OBJECTIVE To explore the in vitro and in vivo inhibitory effects and mechanism of metformin on the malignant biological behavior of esophageal squamous cell carcinoma （ESCC） cells by the hypoxia inducible factor-1α （HIF-1α）/interleukin-8 （IL-8） signaling pathway. METHODS Human ESCC TE1 cells were assigned into blank group， metformin low-， medium-， and high-dose groups （0.5， 1， 2 mmol/L）， IDF-11774 （HIF-1α inhibitor） group （20 μmol/L）， and high-dose metformin+HIF-1α activator dimethyloxalylglycine （DMOG） group. After 24 h treatment， cell proliferation [measured by the positive rate of 5-ethynyl- 2′-deoxyuridine （EdU） and optical density at 450 nm （OD450 value）]， apoptosis， invasion and migration as well as mRNA expressions of proliferating cell nuclear antigen （PCNA）， Bcl-2 interacting mediator of cell death （Bim）， migration and invasion enhancer 1 （MIEN1）， and matrix metalloproteinase-9 （MMP-9）， and protein expressions of HIF-1α and IL-8 in the cells were detected. The xenograft tumor model of nude mice was established. Thirty nude mice were randomly divided into blank group， metformin low-， medium-， and high-dose groups （i.g. administration of metformin 62.5， 125， 250 mg/kg+i.p. administration of equal volume of normal saline）， IDF-11774 group （i.g. administration of 50 mg/kg IDF-11774+i.p. administration of equal volume of normal saline） and high-dose metformin+DMOG group （i.g. administration of metformin 250 mg/kg+i.p. administration of DMOG 250 mg/kg）， with 5 mice in each group. They were given relevant medicine， once a day， for 4 consecutive weeks； the mass and volume of the tumor and protein expressions of HIF-1α and IL-8 in the tumor tissue were determined. RESULTS The EdU positive rate， OD450 value， cell invasion number， scratch healing rate， mRNA expressions of PCNA， MIEN1 and MMP-9， protein expressions of HIF-1α and IL-8， as well as the mass and volume of transplanted tumors and protein expressions of HIF-1α and IL-8 in tumor tissues were decreased by metformin in concentration/dose-dependent manner （P＜0.05）. Additionally，metformin increased the apoptosis rate and mRNA expression of Bim in cells （P＜0.05）. The trend of changes in corresponding indicators in the IDF-11774 group was consistent with that in the metformin groups， whereas DMOG could significantly attenuate the aforementioned effects of high-concentration/high-dose metformin （P＜0.05）. CONCLUSIONS Metformin can inhibit the proliferation， invasion， migration of TE1 cells， and tumor growth of nude mice， and induce cell apoptosis， the mechanism of which may be related to the inhibition of HIF-1α/IL-8 signaling pathway.

ObjectiveTo determine the syndrome of a rat model of follicular dysplasia induced by Tripterygium glycosides based on prescriptions and investigate the mechanism of traditional Chinese medicine intervention via the adenosine monophosphate-activated protein kinase （AMPK）/mammalian target of rapamycin （mTOR）/hypoxia-inducible factor-1 （HIF-1）/vascular endothelial growth factor （VEGF） pathway. MethodForty-eight rats with regular estrous cycles were randomly assigned into a normal group （n=8） and a modeling group （n=40）. The rats in the modeling group were administrated with Tripterygium glycoside suspension （75 mL·kg^-1） by gavage for 30 days. The modeled rats were assigned into model， Siwutang （3.69 g·kg^-1）， Youguiyin （3.11 g·kg^-1）， Zuoguiyin （7.29 g·kg^-1）， and Guishenwan （10.35 g·kg^-1） groups， with 8 rats in each group. The drug intervention lasted for 14 days. The changes of estrous cycle were detected by Pap staining， and a stereoscope was used to observe the morphology of the ovarian tissue. Hematoxylin-eosin staining was employed to observe the pathological changes and follicle count in the ovarian tissue. Enzyme-related immunosorbent assay （ELISA） was used to measure the levels of follicle-stimulating hormone （FSH）， luteinizing hormone （LH）， and estradiol （E₂） in the serum. Real-time fluorescence quantitative polymerase chain reaction and Western blot were employed to determine the mRNA and protein levels， respectively， of AMPK， mTOR， HIF-1， and VEGF in the ovarian tissue. ResultCompared with the normal group， the model group had a disordered estrous cycle， reduced secondary and mature follicles， increased atretic follicles， elevated FSH and LH levels， lowered E₂ level， up-regulated mRNA and protein levels of AMPK， and down-regulated mRNA and protein levels of mTOR， HIF-1， and VEGF （P<0.01）. Compared with the model group， Guishenwan increased secondary and mature follicles， decreased atretic follicles， lowered the FSH and LH levels， elevated the E₂ level， down-regulated the mRNA and protein levels of AMPK， and up-regulated the mRNA and protein levels of mTOR， HIF-1， and VEGF （P<0.01）. Compared with Guishenwan group， Siwutang， Youguiyin， and Zuoguiyin decreased mature follicles， increased atretic follicles （P<0.01）， elevated the LH （P<0.01） and FSH （P<0.05） levels， and lowered the E₂ level （P<0.05）. In addition， Youguiyin up-regulated the protein level of AMPK （P<0.05） and down-regulated the mRNA levels of mTOR and HIF-1 （P<0.01） as well as the mRNA and protein levels of VEGF （P<0.01）. Siwutang down-regulated the mRNA levels of mTOR and HIF-1 as well as the mRNA and protein levels of VEGF （P<0.05）. Zuoguiyin down-regulated the mRNA level of mTOR and the protein and mRNA levels of VEGF （P<0.05）. ConclusionGuishenwan may improve the ovarian function and promote follicle maturation in a rat model of follicular dysplasia by inhibiting the AMPK/mTOR/HIF-1/VEGF pathway， with the therapeutic effect superior to Zuoguiyin， Youguiyin， and Siwutang. It was hypothesized that this model presented the syndrome of kidney-essence deficiency.

This study aimed to investigate the effect of iron overload on the transplant outcomes of pediatric patients with severe aplastic anemia (SAA) undergoing allogeneic hematopoietic stem cell transplantation (allo-HSCT). A retrospective analysis was conducted on the clinical data of 74 children with SAA who received allo-HSCT at the Hematology Department of Wuhan Children’s Hospital between January 2018 and August 2022. Children with iron overload (serum ferritin >1 000 μg/L) before transplantation had a longer disease course, received more red blood cell transfusions, and had a higher number of CD34 + cells infused. Moreover, iron overload significantly delayed the reconstitution of regulatory T cells after transplantation, increasing the incidence of hemorrhagic cystitis and grade Ⅲ-Ⅳ acute graft-versus-host disease after transplantation. However, iron overload did not significantly affect the overall survival and failure-free survival rates of the children.

Objective:To discuss the effect of curcumin on ameliorating doxorubicin(DOX)-induced cytotoxicity in the H9c2 cardiomyocytes,and to clarify its mechanism.Methods:The H9c2 cardiomyocytes were treated with DOX to establish the cardiotoxicity model.The H9c2 cells were divided into normal group,DOX group,DOX+curcumin group,and DOX+curcumin+silent information regulator 3(SIRT3)inhibitor-3(3-TYP)group.After 24 h,the morphology of the cells in various groups were observed;CCK-8 method was used to detect the viabilities of the cells in various groups;TUNEL staining was used to detect the apoptotic rates of the cells in various groups;enzyme-linked immunosorbent assay(ELISA)method was used to detect the activities of catalase(CAT)and superoxide dismutase(SOD)and the levels of malondialdehyde(MDA)in the cells in various groups;2,7-dichlorofluorescein diacetate(DCFH-DA)staining was used to detect the levels of reactive oxygen species(ROS)in the cells in various groups;Western blotting method was used to detect the expression levels of B-cell lymphoma 2(Bcl-2),Bcl-2-associated X protein(Bax),nicotinamide adenine dinucleotide phosphate(NADPH)oxidase 2(NOX2),NADPH oxidase 4(NOX4),SIRT3,acetylated superoxide dismutase 2(Ac-SOD2),and superoxide dismutase 2(SOD2)proteins in the cells in various groups.Results:Compared with normal group,the H9c2 cells in DOX group exhibited swelling,the activity of the cells was decreased(P＜0.05),the CAT and SOD activities were decreased(P＜0.05),the MDA and ROS levels were increased(P＜0.05),the expression levels of NOX2,and NOX4 proteins were increased(P＜0.05),the ratio of Bcl-2/Bax and the expression level of SIRT3 protein were decreased(P＜0.05),and the expression levels of SOD2 and Ac-SOD2 proteins were increased(P＜0.05).Compared with DOX group,the H9c2 cells in DOX+curcumin group showed improved morphology,the activity of the cells was increased(P＜0.05),the CAT and SOD activities were increased(P＜0.05),the MDA and ROS levels were decreased(P＜0.05),the expression levels of NOX2,and NOX4 proteins were decreased(P＜0.05),the ratio of Bcl-2/Bax and the expression level of SIRT3 protein were increased(P＜0.05),and the expression levels of SOD2 and Ac-SOD2 proteins were decreased(P＜0.05).Compared with DOX+curcumin group,the cells in DOX+curcumin+3-TYP group exhibited swelling,the activity of the cells was decreased(P＜0.05),the apoptotic rate of the cells was significantly increased(P＜0.05),the CAT and SOD activities were decreased(P＜0.05),the MDA and ROS levels were significantly increased(P＜0.05),the expression levels of NOX2 and NOX4 proteins were increased(P＜0.05),the ratio of Bcl-2/Bax and the expression level of SIRT3 protein were decreased(P＜0.05),and the expression levels of SOD2 and Ac-SOD2 proteins were increased(P＜0.05).Conclusion:Curcumin suppresses the oxidative stress and apoptosis by activating the SIRT3/SOD2 signaling pathway,improving the cell activity and alleviating the DOX-induced cytotoxicity in the H9c2 cells.

Objective To investigate the clinical and imaging characteristics of myelin oligodendrocyte glycoprotein antibody-associated disease(MOGAD).Methods The clinical symptoms,MRI features,results of laboratory tests and clinical prognosis of 14 MOGAD patients who were hospitalized in our hospital from June 2016 to June 2022 were collected and retrospectively analyzed.Their clinical and imaging characteristics were summarized and discussed.Results Among the 14 enrolled patients,there were 10 males and 4 females,with a male to female ratio of 2.5∶1.Their age of first onset was＜18 years in 3 cases,18～45 years in 8 cases,and＞45 years in 3 cases.Optic neuritis(10/14,71.43％)was the most common clinical type,followed by encephalitis or meningoencephalitis(9/14,64.29％),brainstem encephalitis(5/14,35.71％)and myelitis(5/14,35.71％).Visual impairment(10/14,71.43％)was the most common clinical symptom,followed by headache in 8 cases(8/14,57.14％),fever in 6 cases(6/14,42.86％),dizziness in 6 cases(6/14,42.86％),parethesia in 5 cases(5/14,35.71％),and seizures,limb paralysis,sphincter dysfunction,ataxia,and vomit were all in 4 cases(4/14,28.57％).Four patients(4/14,28.57％)had a history of upper respiratory tract infection before MOGAD onset.There were 10 patients undergoing cerebrospinal fluid(CSF)test,and 8 of them had abnormal results,including 2 patients(2/10,20％)of increased pressure,8 patients(8/10,80％)of larger WBC count in CSF,and 5 patients(5/10,50％)of elevated total protein in CSF.MRI displayed multiple lesion involvement,and there were 7 cases(7/14,50.00％)in cortical/subcortical white matter,6 cases in brainstem(6/14,42.86％),5 cases in optic nerve(5/14,35.71％),4 cases in spinal cord(4/14,28.57％).The hippocampus,thalamus,basal ganglia,and paraventricular white matter were involved in 3 cases(3/14,21.43％),respectively,and the cerebellum and corpus callosum were in 2 cases(2/14,14.29％),respectively.MRI lesions demonstrated patchy hyperintensity on T2 WI and T2 FLAIR,with patchy,nodular and linear enhancement.Among the 10 patients undergoing visual evoked potential(VEP)test,abnormalities were detected in 9 cases(9/10,90％),and 8(8/10,80％)had bilateral visual pathway abnormalities.Eight patients(8/14,57.14％)experienced relapse and remission course.Both methylprednisolone pulse therapy and immunoglobulin modulation therapy were effective in the acute phase.Five patients with relapsed remission presented a significant reduction in recurrence after immunosuppressants.Conclusion MOGAD is manifested with various clinical features,with vision loss,headache,fever and dizziness more common.MRI lesions of MOGAD involve cerebral cortex,subcortical white matter,brainstem,and optic nerve,etc.Patchy hyperintesive signals are observed on T2WI and T2 FLAIR,and some lesions can be enhanced.Corticosteroid pulse therapy and immunoglobulin therapy show effective treatment in the acute phase,and immunosuppressants in the remission phase can reduce relapse.

Objective To observe the clinical effect of Peiyuan Yangxin(cultivating vital essence and nourishing heart)acupuncture and moxibustion method combined with Gubu Pills in the treatment of type 2 diabetes mellitus complicated with osteoporosis(T2DOP).Methods A total of 70 patients with T2DOP were randomly divided into treatment group and control group,with 35 cases in each group.The control group was treated with conventional hypoglycemic and calcium supplementation.On the basis of the control group,the treatment group was treated with Peiyuan Yangxin acupuncture and moxibustion method combined with Gubu Pills for eight weeks.After two months of treatment,the clinical efficacy of the two groups was evaluated,and the changes of serum osteocalcin(OC),total type 1 collagen amino acid extension peptide(PINP),β-collagen special sequence(β-CTX),inorganic phosphorus(P)and glycosylated hemoglobin(HbA1c)were observed before and after treatment.The changes of pain Visual Analogue Scale(VAS),Hamilton anxiety scale(HAMA)and diabetes specific quality of life scale(DSQL)scores were compared before and after treatment between the two groups.The safety and adverse reactions of the two groups were evaluated,and the recurrence rate and rehospitalization rate of the two groups were counted.Results(1)After treatment,the serum OC,PINP,β-CTX and P levels of patients in the two groups were significantly improved(P＜0.05),and the observation group was significantly superior to the control group in improving serum OC,PINP,β-CTX and P levels,with a statistically significant difference(P＜0.05).(2)After treatment,the HbA1c level of patients in both groups was significantly improved(P＜0.05),and the treatment group was significantly superior to the control group in improving HbA1c level,with statistically significant differences(P＜0.05).(3)After treatment,the VAS,HAMA and DSQL scores of patients in the two groups were significantly improved(P＜0.05),and the treatment group was significantly superior to the control group in improving VAS,HAMA and DSQL scores,with statistically significant differences(P＜0.05).(4)The total effective rate was 88.57％(31/35)in the treatment group and 68.57％(24/35)in the control group.The efficacy of the treatment group was superior to that of the control group,and the difference was statistically significant(P＜0.05).(5)The recurrence in the treatment group was significantly less than that in the control group,and the difference was statistically significant(P＜0.05).(6)There were no statistically significant differences of the incidence of adverse reactions between the two groups of patients(P＞0.05).Conclusion Peiyuan Yangxin acupuncture and moxibustion method combined with Gubu Pills in the treatment of T2DOP can significantly reduce the pain and anxiety symptoms of patients,improve the biochemical indexes of bone metabolism,thereby improving the quality of life of patients,increasing the effective rate of treatment,and reducing the recurrence rate and the rate of rehospitalization in the short term.

Objective:To investigate the safety and efficacy of mitoxantrone liposome in the treatment of children with high-risk acute myeloid leukemia(AML).Methods:The children with high-risk AML who received the mitoxantrone liposome regimen at Wuhan Children's Hospital from January 2022 to February 2023 were collected as the observation group,and the children with high-risk AML who received idarubicin regimen were enrolled as controls,and their clinical data were analyzed.Time to bone marrow recovery,the complete remission rate of bone marrow cytology,the clearance rate of minimal residual disease,and treatment-related adverse reactions were compared between the two groups.Results:The patients treated with mitoxantrone liposome showed shorter time to recovery of leukocytes(17 vs 21 day),granulocytes(18 vs 24 day),platelets(17 vs 24 day),and hemoglobin(20 vs 26 day)compared with those treated with idarubicin,there were statistical differences(P＜0.05).The effective rate and MRD turning negative rate in the observation group were 90.9％and 72.7％,respectively,while those in the control group were 94.1％and 76.4％,with no statistical difference(P＞0.05).The overall response rate of the two groups of patients was similar.Conclusion:The efficacy of mitoxantrone liposome is not inferior to that of idarubicin in children with high-risk AML,but mitoxantrone liposome allows a significantly shorter duration of bone marrow suppression and the safety is better.

Objective:To investigate the prognostic value of minimal residual disease(MRD)detected by multi-parameter flow cytometry(MFC)in pediatric patients with acute myeloid leukemia(AML)after induction chemotherapy.Methods:A retrospective study was conducted on 97 pediatric patients initially diagnosed with AML at Wuhan Children's Hospital from August 2015 to December 2022.The study analyzed the results of MRD detection using MFC after the first and second cycles of induction chemotherapy,and its association with prognosis were analyzed.Results:Following the first cycle of induction treatment,57 of the 97 patients tested positive for MRD(MRD1+,58.8％).Subsequently,19 patients remained MRD positive(MRD2+,19.6％)after the second cycle of induction treatment.Kaplan-Meier survival analysis showed that the estimated 3-year overall survival(OS)rate of the 37(64.9％)MRD1+patients who underwent transplantation was significantly higher than that of the 20(35.1％)MRD1+patients who did not undergo transplantation(84.6％vs 40.0％,P=0.0001).Among the 35 MRD1+MRD2-patients,the 3-year OS rate of the 25 children who underwent transplantation was higher than that of the 10 children who did not undergo transplantation(87.2％vs 70.0％,P=0.3229).The 3-year OS rate of the 19 MRD1+MRD2+patients was lower than that of the 35 MRD1+MRD2-patients(57.4％vs 81.8％,P=0.059).In the 19 MRD2+patients,the 3-year OS rate of the 12 children who underwent transplantation was significantly higher than that of the 7 children who did not undergo transplantation(80.8％vs 14.3％,P=0.0007).There was no significant difference in 3-year OS between the 12 MRD1+MRD2+patients and 25 MRD1+MRD2-patients,both treated with transplantation(80.8％vs 87.2％,P=0.8868).In those not treated with transplantation,the 7 MRD1+MRD2+patients had a significantly lower 3-year OS compared with the 10 MRD1+MRD2-patients(14.3％vs 70.7％,P=0.0114).Further multivariate analysis indicated that MRD2 positivity and transplantation were both independent prognostic factors(P=0.031,0.000),while MRD1 positivity was not significantly associated with the overall prognosis of 97 patients(P=0.902).Conclusion:MRD positivity following the second cycle of induction chemotherapy is an independent risk factor for unfavorable outcomes in children with AML.MRD2 positivity indicates a poorer prognosis and can help to identify the candidates requiring transplantation.MRD2 positivity is not a contraindication for transplantation in pediatric patients,and early transplantation significantly improves the prognosis of high-risk patients.

OBJECTIVE: To investigate the clinical features and outcomes of infants (<1 year old) with acute lymphoblastic leukemia (IALL). METHODS: The clinical manifestations, laboratory examination results, treatment and prognosis of 18 infants diagnosed with ALL at our department between January 1, 2014 and August 31, 2022 were retrospectively analyzed. RESULTS: Among the 18 cases of IALL, there were 10 males and 8 females. The median age of patients was 6.5 months old (3 months-11 months old). The median white blood cell count (WBC) was 33.63×10⁹/L ［(3.92-470)×10⁹/L］ at initial diagnosis, including 2 patients with WBC≥300×10⁹/L. Flow cytometric immunophenotyping showed a B-lineage infant ALL in all the 18 patients. Eight of the 18 children had abnormal chromosome karyotype analysis. Fusion gene detection showed 12 KMT2A-rearrangement of 18 patients. 15 patients underwent leukemia related mutation gene screening, among which KRAS, NRAS and FLT3 were the most common mutation genes. 4 patients underwent allogeneic hematopoietic stem cell transplantation and two survived. 14 patients received chemotherapy only and ten survived. The 3-year OS rate was (65.5±11.5)%, while the EFS rate was (46.9±12.3)%. CONCLUSION B-cell ALL and KMT2A rearrangement are prevalent in IALL. The therapeutic effect of IALL with standard childhood ALL protocal is similer to international infant specific protocal.
Male ; Child ; Infant ; Female ; Humans ; Retrospective Studies ; Precursor Cell Lymphoblastic Leukemia-Lymphoma/genetics* ; Prognosis ; Hematopoietic Stem Cell Transplantation ; Mutation

OBJECTIVE: To analyze the gene mutation profile in children with acute lymphocyte leukemia (ALL) and to explore its prognostic significance. METHODS: Clinical data of 249 primary pediatric ALL patients diagnosed and treated in the Department of Hematological Oncology of Wuhan Children's Hospital from January 2018 to December 2021 were analyzed retrospectively. Next-generation sequencing (NGS) was used to obtain gene mutation data and analyze the correlation between it and the prognosis of children with ALL. RESULTS: 227 (91.2%) were B-ALL, 22 (8.8%) were T-ALL among the 249 cases, and 178 (71.5%) were found to have gene mutations, of which 85 (34.1%) had ≥3 gene mutations. NRAS(23.7%), KRAS (22.9%),FLT3(11.2%), PTPN11(8.8%), CREBBP (7.2%), NOTCH1(6.4%) were the most frequently mutated genes, the mutations of KRAS, FLT3, PTPN11, CREBBP were mainly found in B-ALL, the mutations of NOTCH1 and FBXW7 were mainly found in T-ALL. The gene mutation incidence of T-ALL was significantly higher than that of B-ALL (χ²= 5.573，P＜0.05) and were more likely to have co-mutations (P＜0.05). The predicted 4-year EFS rate (47.9% vs 88.5%, P＜0.001) and OS rate (53.8% vs 94.1%, P＜0.001) in children with tp53 mutations were significantly lower than those of patients without tp53 mutations. Patients with NOTCH1 mutations had higher initial white blood cell count （128.64×10⁹/L vs 8.23×10⁹/L，P＜0.001）, and children with NOTCH1 mutations had a lower 4-year EFS rate than those of without mutations (71.5% vs 87.2%, P＝0.037). CONCLUSION Genetic mutations are prevalent in childhood ALL and mutations in tp53 and NOTCH1 are strong predictors of adverse outcomes in childhood ALL, with NGS contributing to the discovery of genetic mutations and timely adjustment of treatment regimens.
Child ; Humans ; Precursor T-Cell Lymphoblastic Leukemia-Lymphoma/genetics* ; Cell Cycle Proteins/genetics* ; Proto-Oncogene Proteins p21(ras)/genetics* ; Retrospective Studies ; Ubiquitin-Protein Ligases/genetics* ; Prognosis ; Precursor Cell Lymphoblastic Leukemia-Lymphoma/genetics* ; Precursor B-Cell Lymphoblastic Leukemia-Lymphoma ; Mutation ; Lymphocytes