1.Allogeneic lung transplantation in miniature pigs and postoperative monitoring
Yaobo ZHAO ; Ullah SALMAN ; Kaiyan BAO ; Hua KUI ; Taiyun WEI ; Hongfang ZHAO ; Xiaoting TAO ; Xinzhong NING ; Yong LIU ; Guimei ZHANG ; He XIAO ; Jiaoxiang WANG ; Chang YANG ; Feiyan ZHU ; Kaixiang XU ; Kun QIAO ; Hongjiang WEI
Organ Transplantation 2026;17(1):95-105
Objective To explore the feasibility and reference value of allogeneic lung transplantation and postoperative monitoring in miniature pigs for lung transplantation research. Methods Two miniature pigs (R1 and R2) underwent left lung allogeneic transplantation. Complement-dependent cytotoxicity tests and blood cross-matching were performed before surgery. The main operative times and partial pressure of arterial oxygen (PaO2) after opening the pulmonary artery were recorded during surgery. Postoperatively, routine blood tests, biochemical blood indicators and inflammatory factors were detected, and pathological examinations of multiple organs were conducted. Results The complement-dependent cytotoxicity test showed that the survival rate of lymphocytes between donors and recipients was 42.5%-47.3%, and no agglutination reaction occurred in the cross-matching. The first warm ischemia times of D1 and D2 were 17 min and 10 min, respectively, and the cold ischemia times were 246 min and 216 min, respectively. Ultimately, R1 and R2 survived for 1.5 h and 104 h, respectively. Postoperatively, in R1, albumin (ALB) and globulin (GLB) decreased, and alanine aminotransferase increased; in R2, ALB, GLB and aspartate aminotransferase all increased. Urea nitrogen and serum creatinine increased in both recipients. Pathological results showed that in R1, the transplanted lung had partial consolidation with inflammatory cell infiltration, and multiple organs were congested and damaged. In R2, the transplanted lung had severe necrosis with fibrosis, and multiple organs had mild to moderate damage. The expression levels of interleukin-1β and interleukin-6 increased in the transplanted lungs. Conclusions The allogeneic lung transplantation model in miniature pigs may systematically evaluate immunological compatibility, intraoperative function and postoperative organ damage. The data obtained may provide technical references for subsequent lung transplantation research.
2.Cost-effectiveness analysis of acquired immunodeficiency syndrome interventions based on Optima HIV model
Yiling ZHENG ; Xin ZHOU ; Yongchun HOU ; Hua CHENG ; Leiming ZHOU ; Zhen NING
Shanghai Journal of Preventive Medicine 2026;38(3):199-205
ObjectiveTo assess the cost-effectiveness of human immunodeficiency virus (HIV) prevention and control strategies across different high-risk populations, investment levels, and allocation proportions in an area, thereby providing a reference for optimizing resource allocation in acquired immunodeficiency syndrome (AIDS) prevention and control. MethodsDemographic, epidemiological, and clinical progression data of the target population in an area from 2018 to 2024 were collected, along with the input costs and intervention coverage of HIV-related projects. The Optima HIV model was utilized to perform fitting and prediction, whereby the allocation of resources to optimized target populations and program interventions was modeled under varying future investment scenarios to predict the impacts on the reduction of new HIV infections and HIV-related deaths. ResultsUnder the scenario of maintaining the current level of intervention input for HIV key populations, new HIV infections and related deaths in the region were predicted to be controlled at a low level by 2030. In terms of intervention input for HIV key populations, it is suggested that appropriately increasing the intervention input for key HIV populations will further reduce new HIV infections and HIV-related deaths in the region. However, when the total input increases to 1.75 times the baseline level, the marginal effect of input will be saturated. Regarding structural adjustments in investment and considering both the current total investment scenario and 1.75 times the total investment scenario, it is predicted that further reductions in regional HIV new infections and HIV-related deaths can be achieved, provided that the intervention input for key populations (including men who have sex with men, MSM) is increased, while concurrently intensifying the proportion of intervention measures such as condom promotion to form optimized intervention portfolios. ConclusionIn the field of HIV/ AIDS prevention and control, sustained commitment to intervention investment, with a strategic focus on interventions for key populations and intensified implementation of critical intervention measures, will effectively improve the epidemiological impacts of HIV/AIDS prevention and control efforts.
3.Surveillance of Oncomelania hupensis snails following interruption of schistosomiasis transmission in Yunnan Province
Siqi NING ; Yi DONG ; Chunhong DU ; Lifang WANG ; Yun ZHANG ; Yuhe HE ; Hua JIANG ; Jiayu SUN ; Chunqiong CHEN ; Jiaqi YAN ; Jihua ZHOU ; Zongya ZHANG ; Hongqiong WANG ; Meifen SHEN ; Jing SONG
Chinese Journal of Schistosomiasis Control 2026;38(2):200-206
Objective To investigate the distribution characteristics of Oncomelania hupensis snails in Yunnan Province fol-lowing interruption of schistosomiasis transmission, so as to provide the evidence for assessing the risk of schistosomiasis transmission and scientifically formulating the schistosomiasis surveillance program. Methods According to the requirements of the National Schistosomiasis Surveillance Scheme (2020 Edition), O. hupensis snail surveillance data were collected from 18 schistosomiasis-endemic counties (cities, districts) in Yunnan Province from 2020 to 2024, including area of snail survey, area of snail habitats, area of re-emerging snail habitats, number of frames surveyed, number of frames with O. hupensis snails, number of O. hupensis snails captured, and number of living snails, and the occurrence of frames with snails and mean density of living snails were calculated. Changes in snail status over the 5-year period from 2020 to 2024 and the differences in snail distributions specified by epidemic intensity, environmental type, and vegetation type were analyzed. Results The areas of snail survey increased from 1 727.96 hm2 in 2020 to 3 894.45 hm2 in 2024 (peak) across 18 schistosomiasis-endemic counties (cities, districts) in Yunnan Province during the period from 2020 through 2024. The areas of snail habitats increased from 70.36 hm2 in 2020 to a peak in 2023 (172.04 hm2), followed by a reduction to 132.36 hm2 in 2024, and the areas of re-emerging snail habitats increased from 42.71 hm2 in 2020 to a peak in 2022 (78.43 hm2), followed by a reduction to 40.21 hm2 in 2024. The occurrence of frames with snails and mean density of living snails increased from 1.24% (3 025/244 404) and (0.033 2 ± 0.038 7) snails/0.1 m2 in 2020 to peaks at 2.03% (6 231/307 563) and (0.066 9 ± 0.068 4) snails/0.1 m2 in 2023, followed by reductions to 1.04% (5 829/559 941) and (0.032 6 ± 0.057 7) snails/0.1 m2 in 2024, respectively. There was a significant difference in the occurrence of frames with snails over the 5-year study period (χ2 = 1 962.95, P < 0.05), and the occurrence of frames with snails reduced by 48.71% in 2024 relative to in 2023 (χ2 = 1 411.05, P < 0.005); however, there was no significant difference in the mean density of living snails over the 5 years (H = 5.310, P > 0.05). There were significant differences in the occurrence of frames with snails (χ2 = 481.27, P < 0.05) and mean density of living snails (H = 6.872, P < 0.05) in schistosomiasis-endemic areas with different epidemic intensities. The occurrence of frames with snails (χ2 = 25.32 and 38.70, both P values < 0.017) and mean density of living snails (Z = 28.55 and 49.96, both P values < 0.017) were higher in schistosomiasis transmission-interrupted and eliminated areas with snails than in schistosomiasis-eliminated areas without snails, and the occurrence of frames with snails (χ2 = 453.54, P < 0.017) and mean density of living snails (Z = −56.97, P < 0.017) were higher in schistosomiasis-eliminated areas with snails than in schistosomiasis transmission-interrupted areas with snails. O. hupensis snails were mainly distributed in paddy fields, dry farmlands and ditches; however, the occurrence of frames with snails (13.40%, 424/3 164) and mean density of living snails [(0.252 8 ± 0.158 7) snails/0.1 m2] were higher in ponds/weirs than in other types of environments (both P values < 0.05). Rice, dry farmland crops and weeds were main vegetations in which O. hupensis snails were distributed, and the occurrence of frames with snails (2.29%, 7 111/310 140) and mean density of living snails [(0.072 3 ± 0.018 9) snails/0.1 m2] were higher in weeds than in other types of environments (both P values < 0.05). Conclusions O. hupensis snails have been effectively controlled in Yunnan Province following implementation of integrated schistosomiasis control measures; however, there are still risk factors for schistosomiasis transmission, including reduced attention to schistosomiasis control and snail re-emergence. Improved control efforts and surveillance system construction and timely identification of risk factors of snail status and timely management are recommended to ensure the achievement of the target of schistosomiasis elimination as scheduled.
4.Rutaecarpine Attenuates Monosodium Urate Crystal-Induced Gouty Inflammation via Inhibition of TNFR-MAPK/NF-κB and NLRP3 Inflammasome Signaling Pathways.
Min LI ; Zhu-Jun YIN ; Li LI ; Yun-Yun QUAN ; Ting WANG ; Xin ZHU ; Rui-Rong TAN ; Jin ZENG ; Hua HUA ; Qin-Xuan WU ; Jun-Ning ZHAO
Chinese journal of integrative medicine 2025;31(7):590-599
OBJECTIVE:
To investigate the anti-inflammatory effect of rutaecarpine (RUT) on monosodium urate crystal (MSU)-induced murine peritonitis in mice and further explored the underlying mechanism of RUT in lipopolysaccharide (LPS)/MSU-induced gout model in vitro.
METHODS:
In MSU-induced mice, 36 male C57BL/6 mice were randomly divided into 6 groups of 8 mice each group, including the control group, model group, RUT low-, medium-, and high-doses groups, and prednisone acetate group. The mice in each group were orally administered the corresponding drugs or vehicle once a day for 7 consecutive days. The gout inflammation model was established by intraperitoneal injection of MSU to evaluate the anti-gout inflammatory effects of RUT. Then the proinflammatory cytokines were measured by enzyme-linked immunosorbent assay (ELISA) and the proportions of infiltrating neutrophils cytokines were detected by flow cytometry. In LPS/MSU-treated or untreated THP-1 macrophages, cell viability was observed by cell counting kit 8 and proinflammatory cytokines were measured by ELISA. The percentage of pyroptotic cells were detected by flow cytometry. Respectively, the mRNA and protein levels were measured by real-time quantitative polymerase chain reaction (qRT-PCR) and Western blot, the nuclear translocation of nuclear factor κB (NF-κB) p65 was observed by laser confocal imaging. Additionally, surface plasmon resonance (SPR) and molecular docking were applied to validate the binding ability of RUT components to tumor necrosis factor α (TNF-α) targets.
RESULTS:
RUT reduced the levels of infiltrating neutrophils and monocytes and decreased the levels of the proinflammatory cytokines interleukin 1β (IL-1β) and interleukin 6 (IL-6, all P<0.01). In vitro, RUT reduced the production of IL-1β, IL-6 and TNF-α. In addition, RT-PCR revealed the inhibitory effects of RUT on the mRNA levels of IL-1β, IL-6, cyclooxygenase-2 and TNF-α (P<0.05 or P<0.01). Mechanistically, RUT markedly reduced protein expressions of tumor necrosis factor receptor (TNFR), phospho-mitogen-activated protein kinase (p-MAPK), phospho-extracellular signal-regulated kinase, phospho-c-Jun N-terminal kinase, phospho-NF-κB, phospho-kinase α/β, NOD-like receptor thermal protein domain associated protein 3 (NLRPS), cleaved-cysteinyl aspartate specific proteinase-1 and cleaved-gasdermin D in macrophages (P<0.05 or P<0.01). Molecularly, SPR revealed that RUT bound to TNF-α with a calculated equilibrium dissociation constant of 31.7 µmol/L. Molecular docking further confirmed that RUT could interact directly with the TNF-α protein via hydrogen bonding, van der Waals interactions, and carbon-hydrogen bonding.
CONCLUSION
RUT alleviated MSU-induced peritonitis and inhibited the TNFR1-MAPK/NF-κB and NLRP3 inflammasome signaling pathway to attenuate gouty inflammation induced by LPS/MSU in THP-1 macrophages, suggesting that RUT could be a potential therapeutic candidate for gout.
Animals
;
NF-kappa B/metabolism*
;
Male
;
Indole Alkaloids/therapeutic use*
;
Signal Transduction/drug effects*
;
Mice, Inbred C57BL
;
Inflammation/complications*
;
Uric Acid
;
Quinazolines/therapeutic use*
;
NLR Family, Pyrin Domain-Containing 3 Protein/metabolism*
;
Humans
;
Gout/chemically induced*
;
Inflammasomes/metabolism*
;
Cytokines/metabolism*
;
THP-1 Cells
;
Mitogen-Activated Protein Kinases/metabolism*
;
Mice
;
Molecular Docking Simulation
;
Lipopolysaccharides
;
Quinazolinones
5.Graph Neural Networks and Multimodal DTI Features for Schizophrenia Classification: Insights from Brain Network Analysis and Gene Expression.
Jingjing GAO ; Heping TANG ; Zhengning WANG ; Yanling LI ; Na LUO ; Ming SONG ; Sangma XIE ; Weiyang SHI ; Hao YAN ; Lin LU ; Jun YAN ; Peng LI ; Yuqing SONG ; Jun CHEN ; Yunchun CHEN ; Huaning WANG ; Wenming LIU ; Zhigang LI ; Hua GUO ; Ping WAN ; Luxian LV ; Yongfeng YANG ; Huiling WANG ; Hongxing ZHANG ; Huawang WU ; Yuping NING ; Dai ZHANG ; Tianzi JIANG
Neuroscience Bulletin 2025;41(6):933-950
Schizophrenia (SZ) stands as a severe psychiatric disorder. This study applied diffusion tensor imaging (DTI) data in conjunction with graph neural networks to distinguish SZ patients from normal controls (NCs) and showcases the superior performance of a graph neural network integrating combined fractional anisotropy and fiber number brain network features, achieving an accuracy of 73.79% in distinguishing SZ patients from NCs. Beyond mere discrimination, our study delved deeper into the advantages of utilizing white matter brain network features for identifying SZ patients through interpretable model analysis and gene expression analysis. These analyses uncovered intricate interrelationships between brain imaging markers and genetic biomarkers, providing novel insights into the neuropathological basis of SZ. In summary, our findings underscore the potential of graph neural networks applied to multimodal DTI data for enhancing SZ detection through an integrated analysis of neuroimaging and genetic features.
Humans
;
Schizophrenia/pathology*
;
Diffusion Tensor Imaging/methods*
;
Male
;
Female
;
Adult
;
Brain/metabolism*
;
Young Adult
;
Middle Aged
;
White Matter/pathology*
;
Gene Expression
;
Nerve Net/diagnostic imaging*
;
Graph Neural Networks
6.Novel biallelic MCMDC2 variants were associated with meiotic arrest and nonobstructive azoospermia.
Hao-Wei BAI ; Na LI ; Yu-Xiang ZHANG ; Jia-Qiang LUO ; Ru-Hui TIAN ; Peng LI ; Yu-Hua HUANG ; Fu-Rong BAI ; Cun-Zhong DENG ; Fu-Jun ZHAO ; Ren MO ; Ning CHI ; Yu-Chuan ZHOU ; Zheng LI ; Chen-Cheng YAO ; Er-Lei ZHI
Asian Journal of Andrology 2025;27(2):268-275
Nonobstructive azoospermia (NOA), one of the most severe types of male infertility, etiology often remains unclear in most cases. Therefore, this study aimed to detect four biallelic detrimental variants (0.5%) in the minichromosome maintenance domain containing 2 ( MCMDC2 ) genes in 768 NOA patients by whole-exome sequencing (WES). Hematoxylin and eosin (H&E) demonstrated that MCMDC2 deleterious variants caused meiotic arrest in three patients (c.1360G>T, c.1956G>T, and c.685C>T) and hypospermatogenesis in one patient (c.94G>T), as further confirmed through immunofluorescence (IF) staining. The single-cell RNA sequencing data indicated that MCMDC2 was substantially expressed during spermatogenesis. The variants were confirmed as deleterious and responsible for patient infertility through bioinformatics and in vitro experimental analyses. The results revealed four MCMDC2 variants related to NOA, which contributes to the current perception of the function of MCMDC2 in male fertility and presents new perspectives on the genetic etiology of NOA.
Humans
;
Male
;
Azoospermia/genetics*
;
Meiosis/genetics*
;
Spermatogenesis/genetics*
;
Adult
;
Exome Sequencing
;
Microtubule-Associated Proteins/genetics*
;
Alleles
;
Infertility, Male/genetics*
7.A propensity score-matched analysis on biopsy methods: enhanced detection rates of prostate cancer with combined cognitive fusion-targeted biopsy.
Bi-Ran YE ; Hui WANG ; Yong-Qing ZHANG ; Guo-Wen LIN ; Hua XU ; Zhe HONG ; Bo DAI ; Fang-Ning WAN
Asian Journal of Andrology 2025;27(4):488-494
The choice of biopsy method is critical in diagnosing prostate cancer (PCa). This retrospective cohort study compared systematic biopsy (SB) or cognitive fusion-targeted biopsy combined with SB (CB) in detecting PCa and clinically significant prostate cancer (csPCa). Data from 2572 men who underwent either SB or CB in Fudan University Shanghai Cancer Center (Shanghai, China) between January 2019 and December 2023 were analyzed. Propensity score matching (PSM) was used to balance baseline characteristics, and detection rates were compared before and after PSM. Subgroup analyses based on prostate-specific antigen (PSA) levels and Prostate Imaging-Reporting and Data System (PI-RADS) scores were performed. Primary and secondary outcomes were the detection rates of PCa and csPCa, respectively. Of 2572 men, 1778 were included in the PSM analysis. Before PSM, CB had higher detection rates for both PCa (62.9% vs 52.4%, odds ratio [OR]: 1.54, P < 0.001) and csPCa (54.9% vs 43.3%, OR: 1.60, P < 0.001) compared to SB. After PSM, CB remained superior in detecting PCa (63.1% vs 47.9%, OR: 1.86, P < 0.001) and csPCa (55.0% vs 38.2%, OR: 1.98, P < 0.001). In patients with PSA 4-12 ng ml -1 (>4 ng ml -1 and ≤12 ng ml -1 , which is also applicable to the following text), CB detected more PCa (59.8% vs 40.7%, OR: 2.17, P < 0.001) and csPCa (48.1% vs 27.7%, OR: 2.42, P < 0.001). CB also showed superior csPCa detection in those with PI-RADS 3 lesions (32.1% vs 18.0%, OR: 2.15, P = 0.038). Overall, CB significantly improves PCa and csPCa detection, especially in patients with PSA 4-12 ng ml -1 or PI-RADS 3 lesions.
Humans
;
Male
;
Prostatic Neoplasms/diagnosis*
;
Propensity Score
;
Retrospective Studies
;
Middle Aged
;
Aged
;
Image-Guided Biopsy/methods*
;
Prostate-Specific Antigen/blood*
;
Prostate/diagnostic imaging*
8.Efficacy and Safety of Decitabine-Based Myeloablative Preconditioning Regimen for allogeneic Hematopoietic Stem Cell Transplantation in Patients with Acute Myeloid Leukemia.
Xia-Wei ZHANG ; Jing-Jing YANG ; Ning LE ; Yu-Jun WEI ; Ya-Nan WEN ; Nan WANG ; Yi-Fan JIAO ; Song-Hua LUAN ; Li-Ping DOU ; Chun-Ji GAO
Journal of Experimental Hematology 2025;33(2):557-564
OBJECTIVE:
To analyze the efficacy and safety of decitabine-based myeloablative preconditioning regimen for allogeneic hematopoietic stem cell transplantation (allo-HSCT) in patients with acute myeloid leukemia (AML).
METHODS:
The clinical characteristics and efficacy of 115 AML patients who underwent allo-HSCT at the First Medical Center of Chinese PLA General Hospital from August 2018 to August 2022 were retrospectively analyzed, including 37 patients treated with decitabine conditioning regimen (decitabine group) and 78 patients without decitabine conditioning regimen (non-decitabine group). The cumulative incidence of relapse (CIR), overall survival (OS), leukemia-free survival (LFS), non-relapse mortality (NRM) and graft versus host disease (GVHD) were analyzed.
RESULTS:
For the patients in first complete remission (CR1) state before allo-HSCT, the 1-year relapse rates of decitabine group(22 cases) and non-decitabine group(69 cases) were 9.1% and 29.6%, respectively, the difference was statistically significant(P =0.042). The 1-year cumulative incidence of acute graft-versus-host disease (aGVHD) in decitabine group and non-decitabine group was 62.2% and 70.5%, respectively, and the 1-year cumulative incidence of chronic inhibitor-versus-host disease (cGVHD) was 18.9% and 14.1%, respectively, there were no significant differences in the incidence of aGVHD and cGVHD between the two groups (P >0.05). Of the 115 patients, there were no significantly differences in the 1-year CIR(21.7% vs 28.8%, P =0.866), NRM(10.9% vs 3.9%, P =0.203), OS(75.2% vs 83.8%, P =0.131) and LFS(74.6% vs 69.1%, P =0.912) between the decitabine group(37 cases) and the non-decitabine group(78 cases).
CONCLUSION
Decitabine-based conditioning regimen could reduce the relapse rate of AML CR1 patients with good safety.
Humans
;
Leukemia, Myeloid, Acute/therapy*
;
Hematopoietic Stem Cell Transplantation/methods*
;
Decitabine/therapeutic use*
;
Transplantation Conditioning/methods*
;
Retrospective Studies
;
Graft vs Host Disease
;
Transplantation, Homologous
;
Male
;
Female
;
Adult
;
Middle Aged
;
Adolescent
;
Young Adult
9.Effective Salvage Mobilization of Peripheral Blood Stem Cells with High-Dose Etoposide in Newly Diagnosed Multiple Myeloma Patients Who Failed Initial Mobilization with High-Dose Cyclophosphamide.
Yue-Qi WANG ; Shi-Hua ZHAO ; Yi MA ; Xi-Lin CHEN ; Shun-Zong YUAN ; Na-Na CHENG ; Guang-Ning SHI ; Wen-Rong HUANG ; Xiu-Bin XIAO
Journal of Experimental Hematology 2025;33(5):1380-1385
OBJECTIVE:
To explore the safety and efficacy of high-dose etoposide (VP-16) combined with recombinant human granulocyte colony-stimulating factor (rhG-CSF) as salvage mobilization for peripheral blood stem cells (PBSC) in newly diagnosed multiple myeloma (NDMM) patients.
METHODS:
From April 2021 to May 2023, eight NDMM patients who had failed to yield sufficient PBSC during initial mobilization with high-dose cyclophosphamide (CTX) combined with rhG-CSF underwent salvage mobilization with 1.2 g/m2 etoposide combined with rhG-CSF 10 μg/(kg·d). The effects and adverse reactions of initial mobilization and salvage mobilization were analyzed.
RESULTS:
For salvage mobilization and initial mobilization, the numbers of PBSC collections were 16 and 18, respectively. The mean value of total collected CD34+ cells were (11.90±5.75)×106/kg and (1.67±0.75)×106/kg (P =0.0010) in salvage mobilization group and initial mobilization group, respectively. The proportion of patients with a total collection of CD34+ cell count≥2×106/kg were 100% and 37.5% (P =0.0625), and the proportion of patients with a total collection of CD34+ cell count≥5×106/kg were 87.5% and 0% (P =0.0156) in salvage mobilization group and initial mobilization group, respectively. For five patients who underwent high-dose CTX initial mobilization but had a total CD34+ cell count < 2×106/kg, successful collection was achieved through salvage mobilization with high-dose VP-16. Salvage mobilization with high-dose VP-16 was scheduled 2-3 weeks after failure of CTX mobilization. Adverse reactions of high-dose VP-16 mobilization did not increase compared to the initial mobilization with high-dose CTX.
CONCLUSION
As a salvage mobilization regimen, VP-16 1.2 g/m2 combined with rhG-CSF is safe and highly effective in NDMM patients who failed to initial mobilization with high-dose CTX combined with rhG-CSF.
Humans
;
Multiple Myeloma/therapy*
;
Etoposide/therapeutic use*
;
Hematopoietic Stem Cell Mobilization/methods*
;
Cyclophosphamide/therapeutic use*
;
Granulocyte Colony-Stimulating Factor
;
Salvage Therapy
;
Peripheral Blood Stem Cells
;
Male
;
Middle Aged
;
Female
;
Peripheral Blood Stem Cell Transplantation
10.Effects of ALKBH5 on the malignant biological behavior of esophageal squamous cell carcinoma and the related mechanism
Peihan MA ; Lingmin ZHANG ; Qian LI ; Ning LU ; Hua WEN ; Mingxin ZHANG
Journal of International Oncology 2025;52(2):79-88
Objective:To investigate the role and potential mechanism of m 6A demethylase ALKBH5 in esophageal squamous cell carcinoma (ESCC) . Methods:Real time fluorogenic quantitative PCR and Western blotting were used to detect ALKBH5 expression in normal esophageal epithelial cells (Het-1A) and ESCC cell lines (Eca109, KYSE30, KYSE150, KYSE410). Transient cell lines with overexpression/knockdown of ALKBH5 (siRNA transfection was divided into si-ALKBH5-1 group and si-ALKBH5-2 group) and control cell lines were constructed. The effects of ALKBH5 on ESCC cell proliferation, migration and apoptosis were studied by MTT assay, cell scratch assay and cell apoptosis assay respectively. The differentially expressed gene was screened by the intersection of RNA sequencing (RNA-seq) and methylated RNA immunoprecipitation sequencing (MeRIP-seq) techniques, and the effect of ALKBH5 on the gene expression was detected by RT-qPCR.Results:Real time fluorogenic quantitative PCR results showed that, the relative expression levels of ALKBH5 RNA in Het-1A, Eca109, KYSE30, KYSE150 and KYSE410 were 1.03±0.28, 0.46±0.02, 0.23±0.10, 0.04±0.02, 0.05±0.00, respectively, with a statistically significant difference ( F=444.60, P<0.001). Western blotting showed that, the relative expression levels of ALKBH5 protein in Het-1A, Eca109, KYSE30, KYSE150 and KYSE410 were 1.14±0.03, 0.88±0.04, 0.66±0.01, 0.69±0.01, 0.95±0.01, respectively, with a statistically significant difference ( F=139.90, P<0.001). MTT test showed that the absorbance ( A) values of KYSE30 control group and ALKBH5 overexpression group were 0.86±0.01 and 1.25±0.01 after 72 hours, respectively, with a statistically significant difference ( t=46.93, P<0.001). The A values of KYSE150 control group and ALKBH5 overexpression group were 1.00±0.03 and 1.43±0.02 after 72 hours, respectively, with a statistically significant difference ( t=16.80, P<0.001). The A values of KYSE30 control group, si-ALKBH5-1 group and si-ALKBH5-2 group were 0.98±0.01, 0.85±0.02 and 0.80±0.09 after 96 hours, respectively, with a statistically significant difference ( F=72.97, P<0.001). The A values of KYSE30 control group were higher than those of si-ALKBH5-1 and si-ALKBH5-2 groups (both P<0.001). The A values of KYSE410 control group, si-ALKBH5-1 group and si-ALKBH5-2 group were 1.28±0.02, 1.15±0.02 and 1.08±0.05 after 72 hours, respectively, with a statistically significant difference ( F=16.97, P=0.003). The A values in KYSE410 control group were higher than those in si-ALKBH5-1 group and si-ALKBH5-2 group ( P=0.020; P=0.003). The cell scratch test showed that 48 hours after scratch, the migration rates of KYSE30 cells in control group and ALKBH5 overexpression group were (27.39±0.54) % and (48.89±5.12) %, respectively, with a statistically significant difference ( t=5.90, P=0.004). The migration rates of KYSE150 cells in control group and ALKBH5 overexpression group were (39.67±0.43) % and (62.20±0.60) %, respectively, with a statistically significant difference ( t=43.15, P<0.001). The migration rates of KYSE30 cells in control group, si-ALKBH5-1 group and si-ALKBH5-2 group were (25.08±1.86) %, (18.75±1.59) % and (7.67±0.52) %, respectively, with a statistically significant difference ( F=74.28, P<0.001). The migration rates of KYSE30 cells in control group were higher than those of si-ALKBH5-1 group and si-ALKBH5-2 group ( P=0.010; P<0.001). The migration rates of KYSE410 cells in control group and si-ALKBH5-1 group, si-ALKBH5-2 group were (38.70±0.41) %, (28.27±1.01) % and (19.40±0.47) %, respectively, with a statistically significant difference ( F=400.20, P<0.001). The migration rates of KYSE410 cells in control group were higher than those of si-ALKBH5-1 group and si-ALKBH5-2 group (both P<0.001). Apoptosis test showed that the apoptosis rates of KYSE30 cells in control group and ALKBH5 overexpression group were (9.59±0.88) % and (4.81±0.89) %, respectively, with a statistically significant difference ( t=6.23, P=0.006). The apoptosis rates of KYSE150 cells in control group and ALKBH5 overexpression group were (8.36±0.09) % and (6.42±0.19) %, respectively, with a statistically significant difference ( t=12.90, P<0.001). The apoptosis rates of KYSE30 cells in control group, si-ALKBH5-1 group and si-ALKBH5-2 group were (4.31±0.19) %, (5.72±0.30) % and (8.94±0.71) %, respectively, with a statistically significant difference ( F=53.46, P<0.001). The apoptosis rates in KYSE30 cells in control group were lower than those in si-ALKBH5-1 group and si-ALKBH5-2 group ( P=0.049; P<0.001). The apoptosis rates of KYSE410 control group, si-ALKBH5-1 group and si-ALKBH5-2 group were (4.45±0.36) %, (5.40±0.11) % and (6.64±0.15) %, respectively, with a statistically significant difference ( F=43.36, P<0.001). The apoptosis rates in KYSE410 cells in control group were lower than those in si-ALKBH5-1 group and si-ALKBH5-2 group ( P=0.016; P<0.001). The differentially expressed gene IGF2BP3 was screened by the intersection of RNA-seq and MeRIP-seq techniques, and the RT-qPCR results showed that, the relative expression levels of IGF2BP3 in KYSE30 were 1.01±0.10 and 1.41±0.10 in control group and ALKBH5 overexpression group, respectively, with a statistically significant difference ( t=4.06, P=0.015). The relative expression levels of IGF2BP3 in KYSE150 were 1.00±0.10 and 1.94±0.24 in control group and ALKBH5 overexpression group, respectively, with a statistically significant difference ( t=5.08, P=0.007). The relative expression levels of IGF2BP3 in KYSE410 were 1.01±0.14, 0.67±0.04 and 0.41±0.04 in control group, si-ALKBH5-1 group and si-ALKBH5-2 group, respectively, with a statistically significant difference ( F=24.36, P=0.001). The relative expression levels of IGF2BP3 in KYSE410 control group were higher than those in si-ALKBH5-1 group and si-ALKBH5-2 group ( P=0.017; P=0.001) . Conclusions:ALKBH5 is underexpressed in ESCC cell lines, but the overexpression of ALKBH5 can promote the proliferation and migration of ESCC cells and inhibit cell apoptosis, which may be related to some negative feedback regulation mechanism. IGF2BP3 may be the downstream target of ALKBH5.

Result Analysis
Print
Save
E-mail