1.Single-cell analysis of immune-lineage features in T-cell large granular lymphocytic leukemia
Ke HUANG ; Lele ZHANG ; Chen QIU ; Ruonan LI ; Yucan SHEN ; Weiwang LI ; Hong PAN ; Zhen GAO ; Liwei FANG ; Yajing CHU ; Weiping YUAN ; Jun SHI
Chinese Journal of Hematology 2025;46(5):453-459
Objective:To investigate alterations in the immune lineage of T-cell large granular lymphocytic leukemia (T-LGLL) at the single-cell transcriptome level and to elucidate its pathogenic mechanisms.Methods:Peripheral blood samples were collected from 5 T-LGLL patients before and after treatment (from June 2019 to December 2020) and 3 healthy controls at the Institute of Hematology & Blood Diseases Hospital, CAMS & PUMC. Single-cell transcriptome sequencing libraries were prepared and sequenced using 10× Genomics technology. Differentially expressed genes in immune cells were compared between patients and healthy donors, followed by pathway enrichment analyses.Results:Profiling 67,237 immune cells revealed that, in T-LGLL: 1) Effector CD8+ T cells exhibited increased numbers, enhanced cytotoxicity, and greater proliferative capacity. Following effective immunosuppressive therapy, both the proliferative capacity and effector functions of these cells significantly decreased ( P<0.05). 2) The proportion of regulatory T (Treg) cells was reduced, accompanied by increased apoptosis. After effective immunosuppressive therapy leading to remission, Treg cell proportions increased, and apoptotic pathways were downregulated ( P<0.05). 3) Antigen-presenting cells (APCs) showed enhanced functionality. Monocytes and dendritic cells were enriched in antigen synthesis and presentation pathways, while B cells displayed increased antigen-binding capacity and were enriched in pathways related to T-cell activation ( P<0.05). 4) Natural killer (NK) cells exhibited attenuated cytotoxic function but demonstrated an enhanced regulatory capacity over T cells ( P<0.05) . Conclusions:T-LGLL patients present a characteristic immunological profile marked by an imbalance in immune homeostasis. This profile includes abnormal activation and expansion of effector CD8 + T cells, and a reduction in Treg cell numbers accompanied by functional impairment. Furthermore, APCs and NK cells were found to positively regulate T-lymphocyte activation, differentiation, and proliferation.
2.Mechanism of microRNA-214-3p targeting CHUK in chemosensitivity of ibrutinib by regulating nuclear factor-κB pathway
Yiqi FANG ; Qianwen BAI ; Xiaodu XU ; Bing WU ; Weiping WU ; Jing QIAN
Journal of Clinical Medicine in Practice 2025;29(11):72-78,84
Objective To investigate the mechanism of microRNA-214-3p(miR-214-3p)targe-ting CHUK to regulate the nuclear factor(NF)-κB pathway in modulating the chemosensitivity of lym-phoma cells to ibrutinib.Methods The half-maximal inhibitory concentration(IC50)was used to verify the ibrutinib-resistant cell model.The expression levels of miR-214-3p and CHUK mRNA in tissues and cells were detected by quantitative real-time polymerase chain reaction(qRT-PCR).Western blotting(WB)was employed to assess CHUK protein levels.A dual-luciferase reporter assay was performed to confirm the direct targeting relationship between miR-214-3p and CHUK.Cell viability was measured using CCK-8 assay.Flow cytometry was used to evaluate cell apoptosis.The expression of the NF-κB p65 signaling pathway was detected by WB.Results The CHUK mRNA and CHUK protein expression levels were higher in ibrutinib-resistant cells than in control cells(P<0.05).The qRT-PCR results showed that miR-214-3p was downregulated in ibrutinib-resistant cells.The dual-luciferase reporter assay confirmed that miR-214-3p directly targeted CHUK.Transfection of miR-214-3p mimics and knockdown of CHUK(si-CHUK)reduced the number of colony-forming cells,whereas transfection of miR-214-3p inhibitor increased the number of colony-forming cells(P<0.05).Transfection of miR-214-3p mimics and knockdown of CHUK(si-CHUK)can promote apoptosis,while transfection of miR-214-3p inhibitor can inhibit apoptosis(P<0.05).Inhibition of the NF-κB p65 pathway was observed in cells transfected with miR-214-3p mimics(P<0.05).Conclusion MiR-214-3p may regulate the NF-κB p65 pathway by targeting the expression of CHUK,thereby enhancing the chemosensitivity of ibrutinib to lymphoma.
3.Comparison of the modified Kadish, AJCC T and Dulguerov T staging systems for olfactory neuroblastoma: analysis of the SEER database
Huanhuan LYU ; Xin WEN ; Jingtao LIN ; Ruihua FANG ; Rui HE ; Mengyu CHEN ; Yihui WEN ; Weiping WEN
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2025;60(6):646-653
Objective:To compare the Kadish T staging, AJCC T staging, and Dulguerov T staging system in terms of their impact on surgical treatment selection and survival prognosis in patients with olfactory neuroblastoma (ONB).Methods:The cases of pathologically confirmed ONB from the Surveillance, Epidemiology, and End Results (SEER) database between 2000 and 2018 were collected and screened. Tumors were staged according to Kadish staging system, AJCC T staging and Dulguerov T staging guidelines. Kaplan-Meier analysis was used to calculate 5-and 10-year overall survival rates for different stages, and the log-rank test was used to detect statistically significant differences. Multivariate analysis was performed using Logistic regression and Cox regression models to explore factors influencing surgical treatment choices and prognosis in ONB patients.Results:A total of 519 ONB patients with complete data available for analysis were included in the study. Multivariate analysis revealed that tumor staging, age, and marital status were closely associated with surgical treatment selection. The 10-year survival rates for patients in stage A, B, and C were 74.1%, 68.7%, 55.0%, respectively. The multivariate analysis failed to show a significant prognostic gradient between adjacent stages in any of the three staging systems.Conclusions:The selection of surgical treatment for ONB is influenced by clinical characteristics such as tumor stage and age. The commonly used Kadish, AJCC T, and Dulguerov T staging systems do not significantly differentiate prognosis between adjacent stages, highlighting the need for the development of a more accurate and comprehensive staging system.
4.Single-cell analysis of immune-lineage features in T-cell large granular lymphocytic leukemia
Ke HUANG ; Lele ZHANG ; Chen QIU ; Ruonan LI ; Yucan SHEN ; Weiwang LI ; Hong PAN ; Zhen GAO ; Liwei FANG ; Yajing CHU ; Weiping YUAN ; Jun SHI
Chinese Journal of Hematology 2025;46(5):453-459
Objective:To investigate alterations in the immune lineage of T-cell large granular lymphocytic leukemia (T-LGLL) at the single-cell transcriptome level and to elucidate its pathogenic mechanisms.Methods:Peripheral blood samples were collected from 5 T-LGLL patients before and after treatment (from June 2019 to December 2020) and 3 healthy controls at the Institute of Hematology & Blood Diseases Hospital, CAMS & PUMC. Single-cell transcriptome sequencing libraries were prepared and sequenced using 10× Genomics technology. Differentially expressed genes in immune cells were compared between patients and healthy donors, followed by pathway enrichment analyses.Results:Profiling 67,237 immune cells revealed that, in T-LGLL: 1) Effector CD8+ T cells exhibited increased numbers, enhanced cytotoxicity, and greater proliferative capacity. Following effective immunosuppressive therapy, both the proliferative capacity and effector functions of these cells significantly decreased ( P<0.05). 2) The proportion of regulatory T (Treg) cells was reduced, accompanied by increased apoptosis. After effective immunosuppressive therapy leading to remission, Treg cell proportions increased, and apoptotic pathways were downregulated ( P<0.05). 3) Antigen-presenting cells (APCs) showed enhanced functionality. Monocytes and dendritic cells were enriched in antigen synthesis and presentation pathways, while B cells displayed increased antigen-binding capacity and were enriched in pathways related to T-cell activation ( P<0.05). 4) Natural killer (NK) cells exhibited attenuated cytotoxic function but demonstrated an enhanced regulatory capacity over T cells ( P<0.05) . Conclusions:T-LGLL patients present a characteristic immunological profile marked by an imbalance in immune homeostasis. This profile includes abnormal activation and expansion of effector CD8 + T cells, and a reduction in Treg cell numbers accompanied by functional impairment. Furthermore, APCs and NK cells were found to positively regulate T-lymphocyte activation, differentiation, and proliferation.
5.Comparison of the modified Kadish, AJCC T and Dulguerov T staging systems for olfactory neuroblastoma: analysis of the SEER database
Huanhuan LYU ; Xin WEN ; Jingtao LIN ; Ruihua FANG ; Rui HE ; Mengyu CHEN ; Yihui WEN ; Weiping WEN
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2025;60(6):646-653
Objective:To compare the Kadish T staging, AJCC T staging, and Dulguerov T staging system in terms of their impact on surgical treatment selection and survival prognosis in patients with olfactory neuroblastoma (ONB).Methods:The cases of pathologically confirmed ONB from the Surveillance, Epidemiology, and End Results (SEER) database between 2000 and 2018 were collected and screened. Tumors were staged according to Kadish staging system, AJCC T staging and Dulguerov T staging guidelines. Kaplan-Meier analysis was used to calculate 5-and 10-year overall survival rates for different stages, and the log-rank test was used to detect statistically significant differences. Multivariate analysis was performed using Logistic regression and Cox regression models to explore factors influencing surgical treatment choices and prognosis in ONB patients.Results:A total of 519 ONB patients with complete data available for analysis were included in the study. Multivariate analysis revealed that tumor staging, age, and marital status were closely associated with surgical treatment selection. The 10-year survival rates for patients in stage A, B, and C were 74.1%, 68.7%, 55.0%, respectively. The multivariate analysis failed to show a significant prognostic gradient between adjacent stages in any of the three staging systems.Conclusions:The selection of surgical treatment for ONB is influenced by clinical characteristics such as tumor stage and age. The commonly used Kadish, AJCC T, and Dulguerov T staging systems do not significantly differentiate prognosis between adjacent stages, highlighting the need for the development of a more accurate and comprehensive staging system.
6.Effect of deliberate practice combined with intensive training in cardiopulmonary resuscitation teaching for nurses receiving standardized training
Weiping WANG ; Qiuying DENG ; Chuanren LING ; Weisheng CHEN ; Haihui FANG ; Liqun HUANG
Chinese Journal of Medical Education Research 2024;23(1):123-127
Objective:To investigate the mid- and long-term training effect of deliberate practice combined with intensive training in cardiopulmonary resuscitation (CPR) teaching for nurses receiving standardized training.Methods:A total of 160 nurses who received the first year of standardized training in Guangdong Provincial Hospital of Traditional Chinese Medicine from July 2020 to March 2021 were enrolled as subjects, and they were randomly divided into experimental group and control group, with 80 nurses in each group. The nurses in the control group received the mode of single deliberate practice, and those in the experimental group received deliberate practice combined with intensive training at months 0, 3, and 6. Assessment was performed for the two groups before and after training,with objective indicators and subjective evaluation as the assessment criteria. Objective score was determined by the immediate objective feedback provided by the electronic display of Resusci Anne ? QCPR, and subjective score was determined by the same group of examiners based on the evaluation of cardiopulmonary resuscitation performed by trainees,including environment evaluation, judgment of consciousness, judgment of respiration, initiation of emergency treatment system, re-evaluation after resuscitation, and overall evaluation. SPSS 18.0 was used for the independent samples t-test, the non-parametric test, and the chi-squares test. Results:After training, there were significant differences between the experimental group and the control group in the objective indicators of the percentage of correct compression (CC%) [85.500 (77.50, 93.00) vs. 81.00 (71.75, 89.00), Z=-2.49, P=0.013] and the proportion of correct ventilation (V%) [84.00 (75.00, 92.75) vs. 80.00 (64.00, 90.25), Z=-2.15, P=0.031]. Both groups had significant improvements in objective and subjective indicators after training (all P<0.05). Conclusions:Deliberate practice combined with intensive training at an interval of 3 months can effectively improve the mid- and long-term CPR skills of nurses receiving standardized training and is more suitable for high-level muscle memory training.
7.Prediction of recurrence risk of estrogen receptor-positive and human epidermal growth factor receptor-2 negative breast cancer using a multi-parameter regression model based on diffusion kurtosis imaging
Weiping ZHOU ; Xingyou ZAN ; Xiao LIU ; Shudong YANG ; Xiangming FANG
Chinese Journal of Radiology 2024;58(2):201-208
Objective:To explore the predictive value of a regression model based on diffusion kurtosis imaging (DKI) parameters for prediction of the recurrence risk in patients with estrogen receptor (ER)-positive, human epidermal growth factor receptor 2 (HER-2)-negative early invasive breast cancer.Methods:A retrospective cross-sectional study was designed. The clinicopathological (age, histological grade, Ki-67 level, etc.) and imaging data of 50 patients (50 lesions) with ER-positive, HER-2 negative early invasive breast cancer who underwent treatment at Wuxi People′s Hospital from January 2016 to December 2018 were retrospectively analyzed. All patients were female, aged 29 to 81 years, and underwent pre-operation conventional MRI and DKI examinations. The volume of breast fibroglandular tissue (FGT), background parenchymal enhancement (BPE), and internal enhancement features were recorded; the peak enhancement (PH), peak enhancement rate, time to peak, mean kurtosis (MK), and mean diffusivity (MD) were calculated. Based on the 21-gene recurrence risk scores, patients were divided into low recurrence risk group and medium-high recurrence risk group. Independent sample t test, Mann-Whitney U test, χ2 test were used to compare the differences of various indicators between the two groups. Two logistic models were constructed with age, PH, MD, and MK as independent variables (Pre1), and with Ki-67, age, PH, MD, and MK as independent variables (Pre2), respectively. The efficacy of the models in predicting low recurrence risk in patients was assessed using receiver operating characteristic curve and area under the curve (AUC). Results:There were 25 cases in the low recurrence risk group and 25 cases in the medium-high recurrence risk group. The differences in age, FGT, PH, MD, MK, and Ki-67 between the low recurrence risk group and the medium-high recurrence risk group were statistically significant (all P<0.05), while other indexes showed no statistically significant differences (all P>0.05). The AUC of Pre1 in predicting low recurrence risk of ER-positive, HER-2 negative early invasive breast cancer was 0.87, with a sensitivity of 0.76 and specificity of 0.88. The AUC of Pre2 for predicting the low recurrence risk of ER-positive, HER-2 negative early invasive breast cancer was 0.92, with a sensitivity of 0.84, and specificity of 0.92. Conclusions:A multi-parameter model based on DKI can effectively predict the recurrence risk of ER-positive and HER-2 negative breast cancer. The model with combination of Ki-67 can further improve the predictive efficacy, and help effectively identify patients at low recurrence risk.
8.Application value of the deep learning-based image reconstruction algorithm in combined head and neck CT angiography with low radiation dose
Yangfei LI ; Weiping ZHU ; Yidi HOU ; Jianxin PANG ; Yicheng FANG ; Huayong ZHU
Chinese Journal of Radiological Medicine and Protection 2024;44(1):53-59
Objective:To explore the differences between the deep learning-based image reconstruction (DLIR) and the adaptive statistical iterative reconstruction V (ASiR-V) algorithms in the radiation dose and image quality of head and neck CT angiography (CTA).Methods:The data of 80 patients undergoing head and neck CTA due to vascular diseases in the head and neck were prospectively collected. These patients were randomly divided into groups A and B based on their examination sequence. The CTA images of group A were reconstructed based on ASiR-V 50%, with a tube voltage of 120 kV and a noise index of 11.0. In contrast, those of group B were reconstructed based on ASiR-V 50% (for group B1) and DLIR-H (for group B2), with a tube voltage of 80 kV and a noise index of 9.0. Then, the radiation doses and image quality of both groups were compared using the independent-sample t-test. The radiation doses, and both subjective and objective image quality of the two imaging method were compared through the Kruskal-Wallis test and the Wilcoxon rank-sum test. The independent- or paired-sample t-test was employed to measure inter-group vascular enhanced CT values, as well as signals and noise from regions of interest (ROIs), with signal-to-noise ratios (SNRs) and contrast-to-noise ratios (CNRs) calculated. Results:The effective doses of groups A and B were (0.77±0.08) and (0.45±0.05) mSv, respectively, with a statistically significant difference ( t = 21.96, P < 0.001). The vascular enhanced CT values, SDs, SNRs, and CNRs in the arch of the aorta, the initial and bifurcation parts of the common carotid artery, and the M1 segment of the middle cerebral artery showed statistically significant differences among groups A, B1, and B2 ( F = 67.69, 68.50, 50.52, 74.10, 63.10, 91.22, 69.16, P < 0.001). Additionally, statistically significant differences were observed in the subjective scores of image quality among groups A, B1, and B2 ( Z = 71.06, P < 0.05). Conclusions:The DLIR algorithm can further reduce the radiation dose in head and neck CTA examination while significantly reducing image noise and ensuring image quality, thus demonstrating high clinical application value.
9.Analysis of Mycoplasma pneumoniae infection in children in a tertiary hospital of Nanjing before and after the COVID-19 out-break
Fang HE ; Xiaoyang ZHANG ; Weiping XU ; Yidi GUO ; Guoying ZHANG
Chinese Journal of Clinical Laboratory Science 2024;42(11):873-876
Objective To investigate the epidemiological characteristics of Mycoplasma pneumoniae(MP)infection in children in Nanjing Integrated Traditional Chinese and Western Medicine Hospital before and after the COVID-19 outbreak,and provide an experi-mental basis for the prevention and treatment of MP infection.Methods The clinical data of 17 976 children visited Nanjing Integrated Traditional Chinese and Western Medicine Hospital from January 2019 to November 2023 due to respiratory tract infections were retro-spectively analyzed.The levels of serum specific MP-IgM in the children were detected by the direct luminescence immunoassay,and the detection rates of MP-IgM in different genders,seasons,and ages before and after the COVID-19 epidemic were analyzed using the chi square test to explore the epidemiological characteristics of MP infection.Results The total detection rate of serum MP-IgM in 17 976 children with respiratory tract infections was 28.45%(5 114/17 976).Among them,the total detection rate of serum MP-IgM in female children(31.69%,2 672/8 432)was significantly higher than that in male children(25.59%,2 442/9 544,χ2=81.89,P<0.001).The detection rate of serum MP-IgM was highest in 2019(34.35%,1 415/4 119),followed by in 2023(30.11%,2 409/8 001)and in 2020-2022(22.03%,1 290/5 856),with statistically significant difference(χ2=19.95,P<0.001).The detection rate of serum MP-IgM was highest in autumn(33.16%,1 683/5 075),followed by in summer(28.61%,1 053/3 681),winter(27.65%,1 826/6 604),and spring(21.10%,552/2 616),with statistically significant difference(χ2=126.90,P<0.001).Among different age groups,the detection rate of serum MP-IgM was highest in the age group of 7-9 years old(35.83%,1 190/3 321),fol-lowed by 4-6 years old(28.06%,1 882/6 707),1-3 years old(26.55%,1 493/5 623),10-18 years old(23.64%,486/2 056),and<1 year old(23.42%,63/269),with statistically significant difference(χ2=126.11,P<0.001).Conclusion MP is a common pathogen of respiratory tract infections in children,especially in the age range of 7-9 years old,with female children having a higher in-cidence than male children,and the peak incidence in autumn.The effective prevention and control measures during the COVID-19 ep-idemic have reduced the detection rate of MP-IgM,which may provide certain experimental basis for the control and prevention of MP infection transmission and other respiratory diseases.
10.The application value of MRI high-definition diffusion weighted imaging combined with T1WI dynamic contrast enhancement in preoperative T-stage of rectal cancer
Hongyan WAN ; Xiangming FANG ; Wei SHEN ; Xiaoyun HU ; Weiping ZHOU ; Zhiqiang TIAN ; Shudong YANG ; Haixia MAO ; Zongming ZHU
Journal of Practical Radiology 2024;40(6):926-930
Objective To explore the effectiveness of high-definition diffusion weighted imaging(DWI)sequence combined with T1 WI-fat suppression(FS)dynamic contrast enhancement(DCE)sequence for preoperative T-stage of rectal cancer by using 3.0T MRI standardized scanning.Methods A retrospective analysis was conducted on MRI images of 57 patients with rectal cancer confirmed by pathology.Before surgery,the patients underwent 3.0T MRI standardized rectal cancer scan methods,including routine sequence,high-definition DWI sequence,and T1 WI-FS DCE sequence,etc.Then two experienced physicians evaluated the T-stage of preoperative rectal cancer through high-definition DWI(transverse and sagittal sections)and T1 WI-FS DCE sequences in the double-blind method.Using the postoperative pathological results of rectal cancer as the"gold standard",two sequences were combined to evaluate the accuracy,sensitivity,and specificity of rectal cancer T-stage.Results Among the 57 cases,there were 9 cases of upper rectal cancer,39 cases of middle rectal cancer,and 9 cases of lower rectal cancer.The accuracy rates of preoperative T-stage diagnosis for rectal cancer by two evaluator were both 85.7%(6/7)in T1 stage,88.2%(15/17)and 94.1%(16/17)in T2 stage,96.9%(31/32)and 93.8%(30/32)in T3 stage,and both 100.0%(1/1)in T4 stage.For evaluator 1,the sensitivity and specificity of the rectal cancer T-stage diagnosis were 96.1%and 83.3%,and for evaluator 2 were 94.1%and 83.3%,respectively.For rectal cancer MRI diagnosis,the accuracy rates and sensitivity were higher when combining the high-definition DWI sequence and T1 WI-FS DCE sequence,compared with a single high-definition DWI sequence or T1 WI-FS DCE sequence,and the difference was statistically significant.The average preoperative apparent diffusion coefficient(ADC)value of rectal cancer was compared between the corresponding postoperative pathological T1 to T4 stage groups,and the difference was statistically significant.Conclusion The combination of high-definition DWI sequence and T1 WI-FS DCE sequence improves the accuracy of rectal cancer T-stage,providing assistance for personalized clinical treatment.

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