1.Stability study of umbilical cord mesenchymal stem cells formulation in large-scale production
Wang-long CHU ; Tong-jing LI ; Yan SHANGGUAN ; Fang-tao HE ; Jian-fu WU ; Xiu-ping ZENG ; Tao GUO ; Qing-fang WANG ; Fen ZHANG ; Zhen-zhong ZHONG ; Xiao LIANG ; Jun-yuan HU ; Mu-yun LIU
Acta Pharmaceutica Sinica 2024;59(3):743-750
Umbilical cord mesenchymal stem cells (UC-MSCs) have been widely used in regenerative medicine, but there is limited research on the stability of UC-MSCs formulation during production. This study aims to assess the stability of the cell stock solution and intermediate product throughout the production process, as well as the final product following reconstitution, in order to offer guidance for the manufacturing process and serve as a reference for formulation reconstitution methods. Three batches of cell formulation were produced and stored under low temperature (2-8 ℃) and room temperature (20-26 ℃) during cell stock solution and intermediate product stages. The storage time intervals for cell stock solution were 0, 2, 4, and 6 h, while for intermediate products, the intervals were 0, 1, 2, and 3 h. The evaluation items included visual inspection, viable cell concentration, cell viability, cell surface markers, lymphocyte proliferation inhibition rate, and sterility. Additionally, dilution and culture stability studies were performed after reconstitution of the cell product. The reconstitution diluents included 0.9% sodium chloride injection, 0.9% sodium chloride injection + 1% human serum albumin, and 0.9% sodium chloride injection + 2% human serum albumin, with dilution ratios of 10-fold and 40-fold. The storage time intervals after dilution were 0, 1, 2, 3, and 4 h. The reconstitution culture media included DMEM medium, DMEM + 2% platelet lysate, 0.9% sodium chloride injection, and 0.9% sodium chloride injection + 1% human serum albumin, and the culture duration was 24 h. The evaluation items were viable cell concentration and cell viability. The results showed that the cell stock solution remained stable for up to 6 h under both low temperature (2-8 ℃) and room temperature (20-26 ℃) conditions, while the intermediate product remained stable for up to 3 h under the same conditions. After formulation reconstitution, using sodium chloride injection diluted with 1% or 2% human serum albumin maintained a viability of over 80% within 4 h. It was observed that different dilution factors had an impact on cell viability. After formulation reconstitution, cultivation in medium with 2% platelet lysate resulted in a cell viability of over 80% after 24 h. In conclusion, the stability of cell stock solution within 6 h and intermediate product within 3 h meets the requirements. The addition of 1% or 2% human serum albumin in the reconstitution diluent can better protect the post-reconstitution cell viability.
2.Epidemiological Survey of Hemoglobinopathies Based on Next-Generation Sequencing Platform in Hunan Province, China.
Hui XI ; Qin LIU ; Dong Hua XIE ; Xu ZHOU ; Wang Lan TANG ; De Guo TANG ; Chun Yan ZENG ; Qiong WANG ; Xing Hui NIE ; Jin Ping PENG ; Xiao Ya GAO ; Hong Liang WU ; Hao Qing ZHANG ; Li QIU ; Zong Hui FENG ; Shu Yuan WANG ; Shu Xiang ZHOU ; Jun HE ; Shi Hao ZHOU ; Fa Qun ZHOU ; Jun Qing ZHENG ; Shun Yao WANG ; Shi Ping CHEN ; Zhi Fen ZHENG ; Xiao Yuan MA ; Jun Qun FANG ; Chang Biao LIANG ; Hua WANG
Biomedical and Environmental Sciences 2023;36(2):127-134
OBJECTIVE:
This study was aimed at investigating the carrier rate of, and molecular variation in, α- and β-globin gene mutations in Hunan Province.
METHODS:
We recruited 25,946 individuals attending premarital screening from 42 districts and counties in all 14 cities of Hunan Province. Hematological screening was performed, and molecular parameters were assessed.
RESULTS:
The overall carrier rate of thalassemia was 7.1%, including 4.83% for α-thalassemia, 2.15% for β-thalassemia, and 0.12% for both α- and β-thalassemia. The highest carrier rate of thalassemia was in Yongzhou (14.57%). The most abundant genotype of α-thalassemia and β-thalassemia was -α 3.7/αα (50.23%) and β IVS-II-654/β N (28.23%), respectively. Four α-globin mutations [CD108 (ACC>AAC), CAP +29 (G>C), Hb Agrinio and Hb Cervantes] and six β-globin mutations [CAP +8 (C>T), IVS-II-848 (C>T), -56 (G>C), beta nt-77 (G>C), codon 20/21 (-TGGA) and Hb Knossos] had not previously been identified in China. Furthermore, this study provides the first report of the carrier rates of abnormal hemoglobin variants and α-globin triplication in Hunan Province, which were 0.49% and 1.99%, respectively.
CONCLUSION
Our study demonstrates the high complexity and diversity of thalassemia gene mutations in the Hunan population. The results should facilitate genetic counselling and the prevention of severe thalassemia in this region.
Humans
;
beta-Thalassemia/genetics*
;
alpha-Thalassemia/genetics*
;
Hemoglobinopathies/genetics*
;
China/epidemiology*
;
High-Throughput Nucleotide Sequencing
3.Clinical Characteristics of 67 Cases of Brucellosis
Guo-fen ZENG ; Jiang-feng ZHUANG ; Liang GAO ; Kun-yu SHEN ; Yang ZHANG ; Gui-hua SU ; Zhi-liang GAO ; Xin SHU
Journal of Sun Yat-sen University(Medical Sciences) 2022;43(2):297-304
ObjectiveTo analyze the clinical characteristics of brucellosis patients from two tertiary hospitals in epidemic and non-epidemic areas. MethodsThe clinical data of 67 brucellosis patients hospitalized in The Affiliated Kashi Hospital of Sun Yat-sen University (Kashi, 53) and The Third Affiliated Hospital of Sun Yat-sen University (Guangzhou, 14) from January 2019 to December 2019 were retrospectively reviewed, including demographic data, medical history, symptoms, signs, laboratory tests and treatment, and the differences between the two groups were compared. ResultsThere were 23 patients in Kashi and 11 patients in Guangzhou with definite epidemiological history, respectively (P = 0.019). Fever, fatigue and arthralgia were the most common symptoms of brucellosis. In addition to anorexia, the onset symptoms of patients in the two hospitals were roughly the same. The incidence of fever, muscle pain and fever peak was similar, but the symptoms of hyperhidrosis, fatigue, arthralgia and rash in Kashi were more common in Guangzhou. There were cases of hepatosplenomegaly in both hospitals, but there was no obvious superficial lymphedema. Complications occurred in 39(73.6%) and 11(78.6%) patients in Kashi and Guangzhou, respectively, with no statistical difference (P =0.971), and hepatitis and bone damage were the most common. The incidence of spondylitis (P =0.042) and neurodegenerative brucellosis (P =0.041) in Guangzhou was higher than that in Kashi, and there was no significant difference in other complications. Of the patients in Kashi, 7.5 % were treated with single antibiotics, and 59.7 % were with dual therapy, which were higher than those in Guangzhou (0, 50.0 %). The proportion of the triple scheme in Guangzhou was higher than that of Kashi (50.0 % vs. 11.3 %). ConclusionsThe clinical characteristics of brucellosis in different regions are generally similar, but there are some differences in epidemiological history, symptoms, complications and treatment options. The characteristics of local cases should be concerned in clinical practice to reduce misdiagnosis of brucellosis.
4.Medium and long-term follow-up of the Pul-Stent in treating postoperative branch pulmonary artery stenosis in children with congenital heart disease.
Xin Yi XU ; Ting Liang LIU ; Ying GUO ; Xu ZHANG ; Yi Bei WU ; Mei Rong HUANG ; Li Jun FU ; Fen LI ; Wei GAO
Chinese Journal of Pediatrics 2022;60(1):20-24
Objective: To evaluate the effectiveness and safety of Pul-Stent as the treatment of postoperative branch pulmonary artery stenosis in children with congenital heart disease. Methods: This was a retrospective study. Thirty-three patients who underwent Pul-Stent implantation in Shanghai Children's Medical Center due to postoperative residual pulmonary artery stenosis from August 2014 to June 2015 were included. The immediate curative effect, follow-up and complications of Pul-Stent implantation were assessed. Comparisons between groups were performed with unpaired Student t test. Results: Pul-Stent implantation of 33 patients (19 males and 14 females) were performed successfully. Thirty-one patients underwent percutaneous stenting, and 2 patients underwent hybrid stenting. A total of 35 Pul-Stents were implanted (19 of model small, 15 of model medium and one of model large), 23 stents were planted in the proximal left pulmonary artery and 12 stents were in the proximal right pulmonary artery. The initial diameter of dilation balloon ranged from 6 to 16 mm, and the long sheath of percutaneous implantation ranged from 8 to 10 F in 29 patients (29/31, 94%). After stenting, the diameter of the narrowest segment of pulmonary artery increased from (4.0±1.7) mm to (9.1±2.1) mm in all patients (t=-21.60, P<0.001). The pressure gradient at the stenosis in 26 patients after biventricular correction decreased from (30.5±12.3) mmHg (1 mmHg=0.133 kPa) to (9.9±9.6) mmHg (t=12.92, P<0.001), and the right ventricular to aortic pressure ratio decreased from 0.57±0.14 to 0.44±0.12 (t=7.44, P<0.001). The pressure of the superior vena cava after stenting in 5 patients after cavopulmonary anastomosis decreased from (17.0±1.9) mmHg to (14.0±0.7) mmHg (t=2.86, P=0.046). Two patients died during reoperation for repairing other cardiac malformations. The remaining 31 patients were clinically stable during the follow-up period of (5.3±1.6) years, and one stent fracture was found on chest X-ray. Cardiac catheterization reexaminations in 16 patients showed that restenosis was found in one stent, while stent position and patency were satisfactory in the remaining stents. Nine children underwent post-dilation without stent fracture, displacement or aneurysm formation. Cardiac tomography showed no stent stenosis, fracture observed, or significant change in diameter of the stent in 8 patients. The inner diameter and pulmonary blood perfusion could not be accurately evaluated due to artifacts by cardiac magnetic resonance imaging in 4 patients. Conclusions: Pul-Stent has good compliance and adequate radial strength, and can dilate further over time to accommodate for somatic growth. It performs safely and effectively in treating post-operative branch pulmonary artery stenosis in children.
Child
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China
;
Female
;
Follow-Up Studies
;
Heart Defects, Congenital/surgery*
;
Humans
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Male
;
Pulmonary Artery/surgery*
;
Retrospective Studies
;
Stenosis, Pulmonary Artery/surgery*
;
Stents
;
Treatment Outcome
;
Vena Cava, Superior
5. Anatomic characteristics of the lacrimal sac in the nasal cavity of recurrent dacryocystitis and its application in endoscopic dacryocystorhinostomy combined with stent implantation
Shou-Kai ZHANG ; Yu-Fen GUO ; Shou-Kai ZHANG ; Qin LIU ; Dan-Ru LIANG ; Yu-Fen GUO
Acta Anatomica Sinica 2021;52(6):945-949
Objective To observe the efficacy of endoscopic nasal dacryocystoma anastomosis combined with stent implantation in the treatment of chronic recurrent dacryocystitis. Methods Thirty patients (30 eyes) with chronic dacryocystitis who had relapsed after traditional endoscopic nasal dacryocystorhinostomy were hospitalized in the Department of Otolaryngology Head and Neck Surgery department of Gansu Provincial People's Hospital from January 2010 to January 2019. Nasal lacrimal anastomosis under endoscope, intraoperative combined stent implantation, 3 months after operation, the lacrimal duct stent was removed, the patient ' s tearing symptoms were observed, the lacrimal duct was flushed to determine the lacrimal duct obstruction, and the follow-up period was 12 months. Results Twelve months of follow-up to 12 months, 14 eyes of 30 patients had no complaints of tearing, tearing, and lacrimal tract flushing; The ostomy fistula was unobstructed under nasal endoscope and the fistula was not significantly reduced. There was no complaint of tears in the eyes, tears overflowed, and the lacrimal duct was flushed, but the fistula opening was reduced.; Four eyes showed granulation hyperplasia next to the fistula, which blocked the fistula again. The overall effective rate was 87%. Conclusion Endoscopic dacryocystorhinostomy combined with stent implantation is an effective method for the treatment of chronic recurrent dacryocystitis with good clinical result.
6. Suitable soil moisture contents for water use efficiency and saponins accumulation in Panax notoginseng
Yu-jie ZHENG ; Peng-guo XIA ; Zong-suo LIANG ; Wei-guo CHAI ; Hong-guang ZHAO ; Jian-fen ZHENG ; Kai-jing YAN
Chinese Herbal Medicines 2021;13(2):267-273
Objective: The moisture content in the soil directly affects the yield and quality of Panax notoginseng, especially at the age of three years old. However, the suitable moisture for the growth of P. notoginseng is unknown. In this study, the effects of different soil moisture on the growth of P. notoginseng were studied. Methods: Four different water treatments (0.45 field capacity (FC), 0.60 FC, 0.70 FC, and 0.85 FC) were set up in Shilin County, Yunnan Province, China. The water consumption and daily dynamic of water consumption were determined daily (from April 21 to October 18, 2012), and the daily dynamic of water consumption under different weather conditions (sunny and rainy) was determined. The transpiration coefficient and water use efficiency were calculated through dry matter accumulation and total water consumption. Accumulation of saponins of roots of P. notoginseng were analyzed by HPLC after treated, and the soil moisture content suitable for the growth of P. notoginseng was estimated by regression fitting of the active ingredient accumulation and the soil moisture content. Results: The water consumption of 0.85 FC, 0.70 FC, 0.60 FC and 0.45 FC were 2.89, 3.68, 3.37 and 2.73 kg/plant per day, respectively. The water consumption of P. notoginseng from June to August was greater than other months. The daily dynamic of water consumption on sunny days and sunny days after rain showed a “double peak” feature, and it showed a “single peak” feature on rainy days. The water uses efficiency (WUE) of 0.85 FC, 0.70 FC, 0.60 FC and 0.45 FC were 2.51, 3.32, 4.59, 3.39 gDW/kg H
7.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
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Aged
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COVID-19/virology*
;
China/epidemiology*
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Comorbidity
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Female
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Humans
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Male
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Middle Aged
;
Retrospective Studies
;
Severity of Illness Index
;
Treatment Outcome
8.The diagnostic value of multimodal transvaginal contrast-enhanced ultrasonography combined with fallopian tube patency
Fen FU ; Qin YE ; Rongxi LIANG ; Jingjing GUO ; Jiajia YANG ; Ensheng XUE ; Xiaoqing FAN
Chinese Journal of Ultrasonography 2020;29(9):781-785
Objective:To investigate the diagnostic value of multimodal transvaginal contrast-enhanced ultrasound combined with fallopian tubal patency.Methods:From November 2017 to November 2019, 212 patients with infertility were treated in the outpatient clinic of the Union Hospital of Fujian Medical University. Multimodal contrast-enhanced ultrasonography was used to analyze the diagnostic efficiency differences of four-dimensional transvaginal contrast-enhanced hysterosalpinx (TVS 4D-HyCoSy, 4D), three-dimensional contrast-enhanced hysterosalpingography (TVS 3D-HyCoSy 3D), two-dimensional contrast-enhanced hysterosalpingography (TVS 2D-HyCoSy 2D), and transvaginal harmonic imaging (TVS harmonic imaging, HI) individually and in different combinations. The diagnostic efficiency of tubal patency was compared between the high seniority group and the low seniority group.Results:Two patients gave up the examination because of pain. A total of 420 salpingography in 210 patients were successful, of which 375 were unobstructed and 45 were blocked (13 on the right, 18 on the left, and 7 on both sides). In high seniority group and low seniority group, the diagnostic efficiencies of different mode combinations on fallopian tubal patency were significantly different ( P<0.01). There were significant differences( P<0.05) between 4D+ 3D+ 2D+ HI group and 4D+ 3D+ 2D group, 4D+ 3D+ 2D group and 4D+ 3D group, 4D+ 3D group and 4D group. There was no significant difference ( P>0.05) in the diagnostic efficiency of fallopian tubal patency between high seniority group and low seniority group. Conclusions:The diagnostic efficiency of multimodal transvaginal contrast-enhanced ultrasound combined is higher than that of single mode, and ultrasound doctors in both high and low seniority groups can effectively diagnose fallopian tubal patency, which has important clinical value in the diagnosis of fallopian tube patency.
9.Nucleocapsid protein from porcine epidemic diarrhea virus isolates can antagonize interferon-λ production by blocking the nuclear factor-κB nuclear translocation.
Ying SHAN ; Zi-Qi LIU ; Guo-Wei LI ; Cong CHEN ; Hao LUO ; Ya-Jie LIU ; Xun-Hui ZHUO ; Xing-Fen SHI ; Wei-Huan FANG ; Xiao-Liang LI
Journal of Zhejiang University. Science. B 2018;19(7):570-580
Porcine epidemic diarrhea virus (PEDV) is a highly infectious pathogen that can cause severe diseases in pigs and result in enormous economic losses in the worldwide swine industry. Previous studies revealed that PEDV exhibits an obvious capacity for modulating interferon (IFN) signaling or expression. The newly discovered type III IFN, which plays a crucial role in antiviral immunity, has strong antiviral activity against PEDV proliferation in IPEC-J2 cells. In this study, we aimed to investigate the effect of PEDV nucleocapsid (N) protein on type III IFN-λ. We found that the N proteins of ten PEDV strains isolated between 2013 and 2017 from different local farms shared high nucleotide identities, while the N protein of the CV777 vaccine strain formed a monophyletic branch in the phylogenetic tree. The N protein of the epidemic strain could antagonize type III IFN, but not type I or type II IFN expression induced by polyinosinic-polycytidylic acid (poly(I:C)) in IPEC-J2 cells. Subsequently, we demonstrated that the inhibition of poly(I:C)-induced IFN-λ3 production by PEDV N protein was dependent on the blocking of nuclear factor-κB (NF-κB) nuclear translocation. These findings might help increase understanding of the pathogenesis of PEDV and its mechanisms for evading the host immune response.
Active Transport, Cell Nucleus
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Animals
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Coronavirus Infections
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immunology
;
veterinary
;
virology
;
Genes, Viral
;
Host-Pathogen Interactions
;
immunology
;
Interferons
;
antagonists & inhibitors
;
biosynthesis
;
genetics
;
Interleukins
;
antagonists & inhibitors
;
biosynthesis
;
genetics
;
NF-kappa B
;
metabolism
;
Nucleocapsid Proteins
;
genetics
;
immunology
;
physiology
;
Porcine epidemic diarrhea virus
;
genetics
;
pathogenicity
;
physiology
;
Promoter Regions, Genetic
;
Swine
;
Swine Diseases
;
immunology
;
virology
10.Trephination mini-craniectomy for traumatic posterior fossa epidural hematomas in selected pediatric patients.
Han-Song SHENG ; Chao-Guo YOU ; Liang YANG ; Nu ZHANG ; Jian LIN ; Fen-Chun LIN ; Mao-De WANG
Chinese Journal of Traumatology 2017;20(4):212-215
PURPOSEPosterior fossa epidural hematomas (PFEDH) are uncommon in children but usually require timely surgical intervention due to the risk of life-threatening brainstem compression. We attempt to make the surgical procedure less invasive by treating selected pediatric patients with trephination mini-craniectomy.
METHODSWe retrospectively reviewed the clinical courses, radiological findings, surgical procedures, and prognoses of the pediatric patients who were treated in our departments for traumatic PFEDH from January 2010 to January 2015.
RESULTSDuring this period, a total of 17 patients were surgically treated for PFEDH and 7 were managed with trephination mini-craniectomy for hematoma evacuation. The outcomes were good in all 7 patients as evaluated with Glasgow Outcome Score. There was no mortality in this series. The on average 30-month clinical follow-up showed that patients experienced satisfactory recoveries without complications.
CONCLUSIONOur results suggest that trephination mini-craniectomy is a safe surgical technique for selected PFEDH patients with moderate hematoma volume and stabilized neurological functions. However, standard craniectomy is recommend when there are rapid deteriorations in patients' neurological functions or the hematomas are large and exerted severe mass effects.

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