1.Clinical Analysis of Cutaneous Chronic Graft-Versus-Host Disease Post-Allogeneic Hematopoietic Stem Cell Transplantation in Childhood.
Yu-Xian WANG ; Hao XIONG ; Zhi CHEN ; Li YANG ; Fang TAO ; Yu DU ; Zhuo WANG ; Ming SUN ; Shan-Shan QI ; Lin-Lin LUO
Journal of Experimental Hematology 2025;33(5):1461-1467
OBJECTIVE:
To investigate the clinical features and risk factors associated with cutaneous chronic graft-versus-host disease (cGVHD) after allogeneic hematopoietic stem cell transplantation (allo-HSCT) in children.
METHODS:
A retrospective analysis was conducted on the clinical data of children who underwent allo-HSCT in the Wuhan Children's Hospital from August 1, 2016, to December 31, 2023, and were regularly followed up for 1 year or more. The differences in clinical features between children with and without cutaneous cGVHD were compared, and the risk factors affecting the occurrence of cutaneous cGVHD were analyzed.
RESULTS:
During the study period, 296 children received allo-HSCT. Until December 31, 2024, follow-up showed that 20 children (6.8%) developed cutaneous cGVHD, which manifested as cutaneous lichenification, hyperpigmentation, keratosis pilaris, sclerotic changes, and hair or nail involvement. According to their skin lesion area and degree of grading, 5 cases were mild, 10 cases were moderate, and 5 cases were severe. Multivariate logistic regression analysis revealed that female donors and previous acute GVHD were risk factors for the development of cutaneous cGVHD after allo-HSCT. All 20 children were treated with glucocorticoid ± calcineurin inhibitors (tacrolimus/cyclosporine) as first-line therapeutic agents. Only 1 child improved after first-line treatment. The remaining 19 children treated with a second-line regimen of combination interventions based on individualized status, including 10 children who could not tolerate hormonotherapy or first-line treatment, and showed no significant improvement after 3 months, as well as 9 children with multi-organ cGVHD. After comprehensive second-line treatment, 17 children showed improvement in cutaneous symptoms. There were 3 deaths, including 1 due to primary disease recurrence and 2 due to pulmonary infections.
CONCLUSION
The skin is the first manifestation and most common organ involved in cGVHD in children. Cutaneous cGVHD severely affects the daily activities of transplanted children and requires prolonged immunosuppressive therapy, but has a favorable prognosis. First-line treatments for adults are not applicable to children who usually require a combination treatment with multiple drugs.
Humans
;
Graft vs Host Disease/etiology*
;
Hematopoietic Stem Cell Transplantation/adverse effects*
;
Retrospective Studies
;
Risk Factors
;
Female
;
Child
;
Skin Diseases/etiology*
;
Chronic Disease
;
Transplantation, Homologous
;
Male
;
Child, Preschool
;
Adolescent
2.Expert consensus on evaluation index system construction for new traditional Chinese medicine(TCM) from TCM clinical practice in medical institutions.
Li LIU ; Lei ZHANG ; Wei-An YUAN ; Zhong-Qi YANG ; Jun-Hua ZHANG ; Bao-He WANG ; Si-Yuan HU ; Zu-Guang YE ; Ling HAN ; Yue-Hua ZHOU ; Zi-Feng YANG ; Rui GAO ; Ming YANG ; Ting WANG ; Jie-Lai XIA ; Shi-Shan YU ; Xiao-Hui FAN ; Hua HUA ; Jia HE ; Yin LU ; Zhong WANG ; Jin-Hui DOU ; Geng LI ; Yu DONG ; Hao YU ; Li-Ping QU ; Jian-Yuan TANG
China Journal of Chinese Materia Medica 2025;50(12):3474-3482
Medical institutions, with their clinical practice foundation and abundant human use experience data, have become important carriers for the inheritance and innovation of traditional Chinese medicine(TCM) and the "cradles" of the preparation of new TCM. To effectively promote the transformation of new TCM originating from the TCM clinical practice in medical institutions and establish an effective evaluation index system for the transformation of new TCM conforming to the characteristics of TCM, consensus experts adopted the literature research, questionnaire survey, Delphi method, etc. By focusing on the policy and technical evaluation of new TCM originating from the TCM clinical practice in medical institutions, a comprehensive evaluation from the dimensions of drug safety, efficacy, feasibility, and characteristic advantages was conducted, thus forming a comprehensive evaluation system with four primary indicators and 37 secondary indicators. The expert consensus reached aims to encourage medical institutions at all levels to continuously improve the high-quality research and development and transformation of new TCM originating from the TCM clinical practice in medical institutions and targeted at clinical needs, so as to provide a decision-making basis for the preparation, selection, cultivation, and transformation of new TCM for medical institutions, improve the development efficiency of new TCM, and precisely respond to the public medication needs.
Medicine, Chinese Traditional/standards*
;
Humans
;
Consensus
;
Drugs, Chinese Herbal/therapeutic use*
;
Surveys and Questionnaires
3.Effects of Hot Night Exposure on Human Semen Quality: A Multicenter Population-Based Study.
Ting Ting DAI ; Ting XU ; Qi Ling WANG ; Hao Bo NI ; Chun Ying SONG ; Yu Shan LI ; Fu Ping LI ; Tian Qing MENG ; Hui Qiang SHENG ; Ling Xi WANG ; Xiao Yan CAI ; Li Na XIAO ; Xiao Lin YU ; Qing Hui ZENG ; Pi GUO ; Xin Zong ZHANG
Biomedical and Environmental Sciences 2025;38(2):178-193
OBJECTIVE:
To explore and quantify the association of hot night exposure during the sperm development period (0-90 lag days) with semen quality.
METHODS:
A total of 6,640 male sperm donors from 6 human sperm banks in China during 2014-2020 were recruited in this multicenter study. Two indices (i.e., hot night excess [HNE] and hot night duration [HND]) were used to estimate the heat intensity and duration during nighttime. Linear mixed models were used to examine the association between hot nights and semen quality parameters.
RESULTS:
The exposure-response relationship revealed that HNE and HND during 0-90 days before semen collection had a significantly inverse association with sperm motility. Specifically, a 1 °C increase in HNE was associated with decreased sperm progressive motility of 0.0090 (95% confidence interval [ CI]: -0.0147, -0.0033) and decreased total motility of 0.0094 (95% CI: -0.0160, -0.0029). HND was significantly associated with reduced sperm progressive motility and total motility of 0.0021 (95% CI: -0.0040, -0.0003) and 0.0023 (95% CI: -0.0043, -0.0002), respectively. Consistent results were observed at different temperature thresholds on hot nights.
CONCLUSION
Our findings highlight the need to mitigate nocturnal heat exposure during spermatogenesis to maintain optimal semen quality.
Humans
;
Male
;
Semen Analysis
;
Adult
;
Sperm Motility
;
Hot Temperature/adverse effects*
;
China
;
Middle Aged
;
Spermatozoa/physiology*
;
Young Adult
4.Isolation,identification,and biological characterization of enterotoxigenic Escherichia coli from a South China tiger
Jing-ru XU ; Zhi-hao ZHU ; Yu-qi LI ; Si-si FAN ; Ya-li KANG ; Yu-bin ZHUO ; Ling-shan HUANG ; Shu-qi QIU ; XUE-YUXI ; Xiao-ping WU ; Yu-ting LIAO ; Wei-ye LIN ; Xiao-ziyi XIAO ; Xue-jin LI ; Teng-teng CHEN ; Xi-pan LIN ; Kai-xiong LIN ; Ke-wei FAN
Chinese Journal of Zoonoses 2025;41(6):567-573
This study was aimed at identifying the pathogenic bacteria responsible for the death of a young tiger at the Fujian Meihua Mountain South China Tiger Breeding Research Institute.Tissue samples from the lungs,liver,and intestines of the deceased tiger were collected,and the bacteria were cultured inasterile environment.The bacterial strains were characterized according to their morphological and molecular biological properties,including assessment of virulence genes and antibiotic resistance genes,mouse lethality tests,and antibiotic susceptibility evaluations.A predominant bacterial strain isolated from the liver of the deceased tiger was identified as enterotoxigenic Escherichia coli(ETEC)strain Tiger22513F.Phylogenetic analysis of the 16S rRNA gene revealed that the Tiger22513F strain exhibited close genetic similarity to the reference strain ETEC(MF919609.1),with 99.9%nucleotide similarity,and resided on the same evolutionary branch.The Tiger22513F strain contained 11 antibiotic resistance genes(tetA,sul1,sul3,cmlA,floR,blaTEM,blaSHV,blaCMY-2,qnrA,qnrS,and qnrD)along with five virulence genes(VT1,fyuA,tsh,iucD,and ST).Mouse lethality tests indicated significant pathogenicity toward mice,affecting primarily the lungs,liver,and intestines.Antibiotic susceptibility testing demonstrated that this strain exhibited resistance to various classes of beta-lactam antibiotics,as well as quinolones and aminoglycosides.This investigation successfully isolated a multi-drug resistant enterotoxigenic Escherichia coli strain with pronounced pathogenicity from the liver of a deceased tiger;thus providing valuable scientific insights for clinical diagnosis,as well as prevention and control measures,against ETEC infections in South China tigers.
5.Correlation of serum interleukin-6 and homocysteine levels, monocyte-to-lymphocyte ratio, and serum lipid levels with ulcerative colitis
Zhi QI ; Jian PANG ; Huiming SHAN ; Hao ZHANG ; Tengqi WANG
Chinese Journal of Primary Medicine and Pharmacy 2025;32(2):239-244
Objective:To investigate the correlation of serum interleukin-6 (IL-6) and homocysteine (Hcy) levels, monocyte-to-lymphocyte ratio (MLR), and serum lipid levels [triglycerides (TG), total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), and high-density lipoprotein cholesterol (HDL-C) levels] with ulcerative colitis.Methods:The clinical data of 98 patients with ulcerative colitis admitted to Bayannur Hospital from November 2021 to November 2023 (observation group) were retrospectively analyzed. Forty-nine healthy individuals who were selected at a 2:1 ratio during the same period were included in the control group. Serum IL-6 and Hcy levels, MLR, and lipid levels were compared between the two groups. The diagnostic efficacy of serum IL-6, Hcy, MLR, and lipid levels for ulcerative colitis was assessed using receiver operating characteristic (ROC) curves. Additionally, Pearson correlation analysis was conducted to analyze correlation of serum IL-6 and Hcy levels, MLR, and lipid levels with ulcerative colitis.Results:In the observation group, serum IL-6 and Hcy levels and MLR were (39.87 ± 12.36) pg/mL, (13.01 ± 3.52) μmol/L, and (0.38 ± 0.12), respectively, all of which were significantly higher than those in the control group [(22.3 ± 3.26) pg/mL, (10.05 ± 3.26) μmol/L, (0.29 ± 0.08), t = 9.77, 4.92, 4.78, all P < 0.05]. In the observation group, serum levels of TG, TC, LDL-C, and HDL-C levels were (1.16 ± 0.32) mmol/L, (4.12 ± 1.15) mmol/L, (2.60 ± 0.75) mmol/L, and (1.02 ± 0.17) mmol/L, respectively, all of which were significantly lower than those in the control group [(1.45 ± 0.41) mmol/L, (4.91 ± 0.99) mmol/L, (3.20 ± 0.71) mmol/L, (1.13 ± 0.16) mmol/L, t = 4.71, 4.11, 4.65, 3.77, all P < 0.05]. ROC curve analysis indicated that the areas under the curve (AUC) for diagnosing ulcerative colitis based on serum levels of IL-6, Hcy, MLR, TG, TC, LDL-C, and HDL-C were 0.957, 0.749, 0.746, 0.732, 0.678, 0.722, and 0.681, respectively. Pearson correlation analysis showed that serum levels of IL-6, Hcy, MLR, TG, TC, LDL-C, and HDL-C were all correlated with the severity of ulcerative colitis in patients ( r = 0.501, 0.615, 0.605, -0.577, -0.542, -0.548, -0.646, all P < 0.05). Additionally, serum levels of IL-6, Hcy, and MLR were negatively correlated with lipid levels ( r = -0.806, -0.801, -0.791, -0.649, -0.728, -0.671, -0.720, -0.655, -0.857, -0.877, -0.889, -0.583, all P < 0.05). Conclusions:In patients with ulcerative colitis, serum levels of IL-6, Hcy, and MLR are elevated, while lipid levels are decreased. Additionally, serum levels of IL-6, Hcy, MLR, and lipid levels are associated with the severity of the disease. There is also a correlation between serum levels of IL-6, Hcy, MLR, and lipid levels.
6.Isolation,identification,and biological characterization of enterotoxigenic Escherichia coli from a South China tiger
Jing-ru XU ; Zhi-hao ZHU ; Yu-qi LI ; Si-si FAN ; Ya-li KANG ; Yu-bin ZHUO ; Ling-shan HUANG ; Shu-qi QIU ; XUE-YUXI ; Xiao-ping WU ; Yu-ting LIAO ; Wei-ye LIN ; Xiao-ziyi XIAO ; Xue-jin LI ; Teng-teng CHEN ; Xi-pan LIN ; Kai-xiong LIN ; Ke-wei FAN
Chinese Journal of Zoonoses 2025;41(6):567-573
This study was aimed at identifying the pathogenic bacteria responsible for the death of a young tiger at the Fujian Meihua Mountain South China Tiger Breeding Research Institute.Tissue samples from the lungs,liver,and intestines of the deceased tiger were collected,and the bacteria were cultured inasterile environment.The bacterial strains were characterized according to their morphological and molecular biological properties,including assessment of virulence genes and antibiotic resistance genes,mouse lethality tests,and antibiotic susceptibility evaluations.A predominant bacterial strain isolated from the liver of the deceased tiger was identified as enterotoxigenic Escherichia coli(ETEC)strain Tiger22513F.Phylogenetic analysis of the 16S rRNA gene revealed that the Tiger22513F strain exhibited close genetic similarity to the reference strain ETEC(MF919609.1),with 99.9%nucleotide similarity,and resided on the same evolutionary branch.The Tiger22513F strain contained 11 antibiotic resistance genes(tetA,sul1,sul3,cmlA,floR,blaTEM,blaSHV,blaCMY-2,qnrA,qnrS,and qnrD)along with five virulence genes(VT1,fyuA,tsh,iucD,and ST).Mouse lethality tests indicated significant pathogenicity toward mice,affecting primarily the lungs,liver,and intestines.Antibiotic susceptibility testing demonstrated that this strain exhibited resistance to various classes of beta-lactam antibiotics,as well as quinolones and aminoglycosides.This investigation successfully isolated a multi-drug resistant enterotoxigenic Escherichia coli strain with pronounced pathogenicity from the liver of a deceased tiger;thus providing valuable scientific insights for clinical diagnosis,as well as prevention and control measures,against ETEC infections in South China tigers.
7.Correlation of serum interleukin-6 and homocysteine levels, monocyte-to-lymphocyte ratio, and serum lipid levels with ulcerative colitis
Zhi QI ; Jian PANG ; Huiming SHAN ; Hao ZHANG ; Tengqi WANG
Chinese Journal of Primary Medicine and Pharmacy 2025;32(2):239-244
Objective:To investigate the correlation of serum interleukin-6 (IL-6) and homocysteine (Hcy) levels, monocyte-to-lymphocyte ratio (MLR), and serum lipid levels [triglycerides (TG), total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), and high-density lipoprotein cholesterol (HDL-C) levels] with ulcerative colitis.Methods:The clinical data of 98 patients with ulcerative colitis admitted to Bayannur Hospital from November 2021 to November 2023 (observation group) were retrospectively analyzed. Forty-nine healthy individuals who were selected at a 2:1 ratio during the same period were included in the control group. Serum IL-6 and Hcy levels, MLR, and lipid levels were compared between the two groups. The diagnostic efficacy of serum IL-6, Hcy, MLR, and lipid levels for ulcerative colitis was assessed using receiver operating characteristic (ROC) curves. Additionally, Pearson correlation analysis was conducted to analyze correlation of serum IL-6 and Hcy levels, MLR, and lipid levels with ulcerative colitis.Results:In the observation group, serum IL-6 and Hcy levels and MLR were (39.87 ± 12.36) pg/mL, (13.01 ± 3.52) μmol/L, and (0.38 ± 0.12), respectively, all of which were significantly higher than those in the control group [(22.3 ± 3.26) pg/mL, (10.05 ± 3.26) μmol/L, (0.29 ± 0.08), t = 9.77, 4.92, 4.78, all P < 0.05]. In the observation group, serum levels of TG, TC, LDL-C, and HDL-C levels were (1.16 ± 0.32) mmol/L, (4.12 ± 1.15) mmol/L, (2.60 ± 0.75) mmol/L, and (1.02 ± 0.17) mmol/L, respectively, all of which were significantly lower than those in the control group [(1.45 ± 0.41) mmol/L, (4.91 ± 0.99) mmol/L, (3.20 ± 0.71) mmol/L, (1.13 ± 0.16) mmol/L, t = 4.71, 4.11, 4.65, 3.77, all P < 0.05]. ROC curve analysis indicated that the areas under the curve (AUC) for diagnosing ulcerative colitis based on serum levels of IL-6, Hcy, MLR, TG, TC, LDL-C, and HDL-C were 0.957, 0.749, 0.746, 0.732, 0.678, 0.722, and 0.681, respectively. Pearson correlation analysis showed that serum levels of IL-6, Hcy, MLR, TG, TC, LDL-C, and HDL-C were all correlated with the severity of ulcerative colitis in patients ( r = 0.501, 0.615, 0.605, -0.577, -0.542, -0.548, -0.646, all P < 0.05). Additionally, serum levels of IL-6, Hcy, and MLR were negatively correlated with lipid levels ( r = -0.806, -0.801, -0.791, -0.649, -0.728, -0.671, -0.720, -0.655, -0.857, -0.877, -0.889, -0.583, all P < 0.05). Conclusions:In patients with ulcerative colitis, serum levels of IL-6, Hcy, and MLR are elevated, while lipid levels are decreased. Additionally, serum levels of IL-6, Hcy, MLR, and lipid levels are associated with the severity of the disease. There is also a correlation between serum levels of IL-6, Hcy, MLR, and lipid levels.
8.Relationships between Molecular Genetics and Clinical Features of Children with Acute Mveloid Leukemia
Fei LONG ; Hao XIONG ; Li YANG ; Ming SUN ; Zhi CHEN ; Wen-Jie LU ; Shan-Shan QI ; Fang TAO ; Lin-Lin LUO ; Jing-Pei CHEN
Journal of Experimental Hematology 2025;33(1):69-74
Objective:To analyze the molecular genetic spectrum of children with acute myeloid leukemia(AML),and explore its correlation with clinical characteristics and prognosis.Methods:The clinical and molecular genetic data of 116 children with newly diagnosed AML in Wuhan Children's Hospital from September 2015 to August 2022 were retrospectively analyzed.The Fisher's exact test was used to analyze the correlation of gene mutations with clinical features,and Kaplan-Meier curve was used to analyze the influences of gene mutations on the prognosis.Results:NRAS(22%),KRAS(14.9%),and KIT(14.7%)mutations were the most common genetic abnormalities in 116 children with AML.Children with KIT,CEBPA and GATA2 mutations showed a higher median onset-age than those without mutations(all P<0.05).Children with FLT3-ITD mutation exhibited a higher white blood cell count at initial diagnosis compared to those without mutations(P<0.05).Children with ASXL2 mutation had lower platelet count and hemoglobin at initial diagnosis than those without mutations(both P<0.05).KIT mutations were often co-occurred with t(8;21)(q22;q22).There was no significant relationship between gene mutation and minimal residual disease(MRD)remission rate after the first and second induction therapy(P>0.05).KIT and NRAS mutations were not associated with prognosis significantly(P>0.05).The overall survival(OS)rates of children with CEBPA and FLT3-ITD mutations were superior to those without mutations,but the differences were not statistically significant(P>0.05).The 3-year OS rate of 61 children treated by allogeneic hematopoietic stem cell transplantation was 89.8%,which was significantly higher than 55.2%of those only treated by chemotherapy(P<0.001).Conclusions:Gene mutations are common in children with AML,and next-generation sequencing can significantly improve the detection rate of gene mutations,which can guide the risk stratification therapy.In addition,FLT3-ITD and KIT mutations may no longer be poor prognostic factors.
9.Bioequivalence study of olmesartan medoxomil tablet in Chinese healthy subjects
Na SHAN ; Da-Hai JIANG ; Lin-Lin MIAO ; Zhen-Li REN ; Peng-Bo JIN ; Pei-Qi HAO ; Li AN ; Hong ZHU ; Yong XIN ; Guang-De YANG ; Feng LIU
The Chinese Journal of Clinical Pharmacology 2024;40(20):3033-3037
Objective To study the bioequivalence of test and reference olmesartan tablet in Chinese healthy subjects after single dose under fasting and fed conditions.Methods A single-center,random,open,single-dose,two-preparations,double-period,crossover study was adopted.A total of 48 healthy adult male and female subjects(24 cases of fasting test and 24 cases of fed test)were included in the random crossover administration.Single oral dose 20 mg of test and reference were taken under fasting and postprandial conditions,respectively.Plasma concentration of olmesartan in plasma were determined by liquid chromatography tandem mass spectrometry.The main pharmacokinetic parameters were calculated by Phoenix WinNonlin 8.0 software.Results The main pharmacokinetic parameters of the test and reference preparations of olmesartan tablets in the fasting group were as follows:Cmax were(653.06±133.53)and(617.37±151.16)ng·mL-1,AUC0-t were(4 201.18±1 035.21)and(4 087.38±889.99)ng·mL-1·h,AUC0-∞ were(4 254.30±1 058.90)and(4 135.69±905.29)ng·mL-1·h.The main pharmacokinetic parameters of the test and reference preparations of olmesartan tablets in the postprandial group were as follows:Cmax were(574.78±177.05)and(579.98±107.74)ng·mL-1,AUC0-t were(3 288.37±866.06)and(3 181.51±801.06)ng·mL-1·h,AUC0-∞ were(3 326.11±874.26)and(3 242.01±823.09)ng·mL-1·h.Under fasting and postprandial conditions,the 90%confidence intervals of the main pharmacokinetic parameters of the test and reference preparations are both 80.00%-125.00%.Conclusion Under fasting and postprandial conditions,a single oral dose of test and reference preparations olmesartan tablets in Chinese healthy adult volunteers showed bioequivalence.
10.Correlation between serum uric acid level and body composition, exercise capacity and cardiopulmonary function in medical examination population
Shan LIU ; Jia CUI ; Wei ZHAO ; Honghai HE ; Jie GE ; Xiaoyan HAO ; Tianyi QI ; Peng WANG
Chinese Journal of Health Management 2024;18(1):24-28
Objective:To investigate the correlation between blood uric acid level and body composition, exercise capacity, and cardiopulmonary function in medical examination population.Methods:In this cross-sectional study, 83 individuals who underwent physical examinations at Peking University Third Hospital from June 1, 2023, to October 1, 2023, and met the inclusion criteria were included. According to whether they had hyperuricemia (HUA), the participants were divided into HUA group (53 cases) and non-HUA group (30 cases). Body composition parameters, such as body mass index and visceral fat area, were measured with a body composition analyzer. Exercise capacity indicators, including grip strength, vertical jump, back strength, and sit-and-reach test, were measured using specific monitoring devices. Cardiopulmonary function was assessed using the stair index test. The clinical characteristics of the two groups were compared with t-tests or chi-square tests, and the correlation between uric acid levels and body composition, exercise capacity, and cardiopulmonary function was analyzed. Results:The HUA group had significantly higher skeletal muscle mass, body fat mass, body mass index, and visceral fat area when compared with the non-HUA group [(31.92±5.60) vs (26.11±6.19) kg, (23.66±9.33) vs (17.19±5.00) kg, (26.53±3.68) vs (23.27±3.59) kg/m2, 91.20 (74.25, 123.90) vs 68.25 (56.25, 90.48) cm 2, respectively] (all P<0.05). The grip strength, vertical jump, and back pull strength were all lower in the HUA group [32.70 (25.25, 40.30) vs 42.35 (35.95, 48.10) kg, 30.30 (24.10, 36.48) vs 40.55 (33.06, 45.10) kg, 24.20(20.60, 32.23) vs 29.90 (25.20, 35.50) cm, 65.60 (51.75, 78.00) vs 91.00 (67.25, 111.50) kg, respectivley] (all P<0.05). The increased step index was positively correlated with reduced risk of hyperuricemia ( OR=0.875, 95% CI: 0.793-0.966) ( P<0.05). Conclusions:Blood uric acid level is correlated with cardiopulmonary function in medical examination population. Individuals with better cardiopulmonary function have a lower risk of developing HUA. However, the relationship between blood uric acid level and body composition and exercise capacity is not clear.

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