1.Artemin promotes proliferation and invasion of malignant peripheral nerve sheath tumor cells through the PI3K/Akt pathway
Hongliang ZHANG ; Haotian LIU ; Junyang LIU ; Chao ZHANG ; Ting LI ; Zhichao LIAO ; Yancheng LIU ; Jingyu ZHANG ; Kai ZHU ; Shuang LI ; Jinwei LIU ; Jilong YANG
Chinese Journal of Oncology 2025;47(2):149-159
Objective:To investigate the expression of Artemin (ARTN) in malignant peripheral nerve sheath tumor (MPNST), its effect on the malignant behavior of MPNST cells, and its signaling pathway.Methods:Fifty-one MPNST paraffin embedded tissues through surgical resection at Tianjin Medical University Cancer Hospital from January 1995 to November 2011 were collected, the expression of the ARTN protein was detected by immunohistochemistry, and the relationship between the ARTN protein expression and the clinical pathological characteristics and prognosis were analyzed. In human MPNST cell lines ST-8814 (NF-1) and STS26T(sporadic), ARTN overexpression and low expression cell lines were constructed by transfecting ARTN overexpression plasmids and ARTN small interfering RNA (siRNA), respectively. The expression of ARTN mRNA was detected by real time quantitative polymerase chain reaction (RT-qPCR), the expression of the ARTN protein and Phosphoinositide 3-kinase(PI3K)/Akt signaling pathway related proteins were detected by Western blot. CCK-8 assay was used to detect cell proliferation ability, and cell invasion assay was used to detect cell invasion ability. The pathway proteins that interacted with ARTN were searched in the STRING database, and the functional pathways were clarified by Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis. The PI3K/Akt pathway specific inhibitor LY294002 was used to block the PI3K/Akt pathway of ST-8814 and STS26T cells to observe the changes in cell proliferation and invasion.Results:Among the 51 MPNST tissue specimens, 22 cases showed a high expression of the ARTN protein and 29 cases showed a low expression of the protein. Higher expressions of the ARTN protein was associated with larger tumor diameters and disease progression (recurrence or metastasis) (both P<0.05). The median disease-free survival (DFS) of patients with a low expression of the ARTN protein was 26.2 months, and the median overall survival (OS) was 66.9 months. The median DFS and median OS of patients with a high expression of the ARTN protein were 10.7 months and 53.8 months, respectively. The log rank test results showed that the progression free survival rate of patients with a high expression of the ARTN protein was worse than that of patients with a low expression ( P=0.027), but the difference in overall survival rate between the two groups was not statistically significant ( P=0.790), which was also confirmed by Cox regression analysis. The CCK-8 assay results showed that after 48 hours of transfection, the absorbance ( A) values of ST-8814 and STS26T cells in the ARTN overexpression group were 1.35±0.01 and 1.10±0.02, respectively, which were higher than those in the empty plasmid control group (1.05±0.01 and 0.78±0.01, both P<0.01), while the A values of ST-8814 and STS26T cells in the ARTN siRNA group were 0.35±0.01 and 0.61±0.01, respectively, which were lower than those in the control siRNA group (0.74±0.01 and 1.10±0.04, both P<0.01). The results of cell invasion assay showed that the number of transmembrane cells in ST-8814 and STS26T cells overexpressing ARTN was (29.67±2.08) and (31.67±2.08), respectively, which were higher than those in the empty plasmid control group [(20.00±1.00) and (24.33±1.15), both P<0.01]. The number of transmembrane cells in ST-8814 and STS26T cells in the ARTN siRNA group were (14.00±2.00) and (19.33±1.53), respectively, which were lower than those in the control siRNA group [(19.33±2.52) and (23.33±0.58), both P<0.05].The KEGG results showed that ARTN is associated with multiple tumor signaling pathways, especially the PI3K/Akt signaling pathway. Western blot results showed that overexpression of ARTN upregulated the expression of p-PI3K and p-Akt proteins in ST-8814 and STS26T cells (both P<0.01).After knocking down ARTN expression, the expression of p-PI3K and p-Akt proteins was significantly down regulated (both P<0.01). LY294002 could significantly inhibit the effect of ARTN overexpression on ST-8814 and STS26T cells after blocking the PI3K/Akt pathway. The A values of ST-8814 and STS26T cells in the ARTN overexpression+LY294002 group were 1.09±0.06 and 0.82±0.01, respectively, which were lower than those in the ARTN overexpression group (1.50±0.01 and 1.29±0.01, respectively, both P<0.01). The numbers of transmembrane cells in the cell invasion assay were 16.67±3.21 and 19.67±2.31, respectively, which were also lower than those in the ARTN overexpression group (29.67±2.08 and 31.67±2.08, respectively, both P<0.01). Conclusions:In MPNST, a high expression of the ARTN protein was associated with larger tumor size, disease progression, and worse DFS. ARTN promotes the proliferation and invasion of MPNST cells through the PI3K/Akt signaling pathway.
2.Efficacy and safety of chimeric antigen receptor T cell therapy combined with zanubrutinib in the treatment of relapsed/refractory diffuse large B-cell lymphoma.
Langqi WANG ; Chunyan YUE ; Xuan ZHOU ; Jilong YANG ; Bo JIN ; Bo WANG ; Minhong HUANG ; Huifang CHEN ; Lijuan ZHOU ; Sanfang TU ; Yuhua LI
Chinese Medical Journal 2025;138(6):748-750
3.Analyzing the influencing factors of work-related musculoskeletal disorders in bus drivers
Chunshuo CHEN ; Xiongda HE ; Bin XIAO ; Xiaming CHEN ; Junle WU ; Jilong YANG ; Yongjian JIANG ; Yanhui LAN ; Maosheng YAN ; Haihua BIN
China Occupational Medicine 2025;52(6):624-630
Objective To investigate the prevalence and influencing factors of work-related musculoskeletal disorders (WMSDs) among bus drivers. Methods A total of 962 drivers from a bus company in Shenzhen City were selected as the research subjects using the judgment sampling method. The Musculoskeletal Disorders Questionnaire for Bus Drivers was used to investigate the prevalence of WMSDs among the research subjects. Results The prevalence of WMSDs was 37.8% in the bus drivers. The prevalence of WMSDs was higher in the low back/waist, neck, and shoulder compared with other body parts, with prevalence of 24.0%, 20.2%, and 14.8%, respectively. The prevalence of single-site and multi-site WMSDs was 18.5% and 19.3%, respectively. The results of the multivariable logistic regression analysis showed that longer job tenure and higher alcohol consumption frequency were associated with higher WMSDs risks (all P<0.01). Weekly work time >48 hours, insufficient rest, work-related fatigue, uncomfortable auxiliary lenses, non-upright trunk posture, prolonged static trunk posture, prolonged wrist flexion, and habitual staying up late were risk factors of WMSDs in the bus drivers (all P<0.05). Conclusion The prevention and treatment of WMSDs among the bus drivers cannot be ignored. Personal characteristics, work organization, work environment, working posture and sleeping habits are the factors that influence the development of WMSDs.
4.Effects of electroacupuncture at "Shenting", "Benshen" and "Baihui" acupoints on mechanical pain threshold and PI3K/TRPV1 pathway in trigeminal ganglion in rats with infraorbital nerve chronic constriction injury
Qiuhui SUN ; Yang GAO ; Jilong WANG ; Yingzhe SUN ; Tianyang YU ; Yuanzheng SUN
International Journal of Traditional Chinese Medicine 2025;47(4):488-493
Objective:To observe the effects of electroacupuncture at "Shenting", "Benshen" and "Baihui" acupoints on the mechanical pain threshold and JAK/PI3K/TRPV1 pathway in the trigeminal ganglion of rats with trigeminal neuralgia model; To explore the related mechanism.Methods:Totally 36 male SD rats were divided into sham-operation group, model group and electroacupuncture group using random number table method, with 12 rats in each group. Except for the sham-operation group, rats in the model group and electroacupuncture group were modeled by infraorbital nerve chronic constriction injury. In the electroacupuncture group, electroacupuncture was performed at "Shenting", "Benshen" and "Baihui" 14 days after surgery, 20 min every day, once every other day, and every 3 times for 1 course of treatment with an interval of 2 d between each course of treatment. A total of 2 courses of treatment were performed. VonFrey fiber wire was used to measure the mechanical pain threshold of rat whisker pads. HE staining was used to observe the morphology and structure of trigeminal ganglion of rats in each group. Immunohistochemistry and Western blot method were used to observe the protein expressions of JAK, PI3K and TRPV1 in trigeminal ganglion of rats, and the serum level of IL-6 was detected in the serum of rats by ELISA.Results:Compared with the model group, the pain threshold of the electroacupuncture group increased significantly ( P<0.05), and the infiltration of inflammatory cells and demyelination in the trigeminal nerve ganglion decreased, and the positive expressions of JAK, PI3K, and TRPV1 in the trigeminal ganglion decreased ( P<0.05 or P<0.01), the protein expressions of JAK2, PI3K, and TRPV1 decreased ( P<0.05 or P<0.01), and the serum IL-6 level decreased ( P<0.01). Conclusions:Electroacupuncture at "Shenting", "Benshen" and "Baihui" may play an analgesic role by regulating IL-6 levels and inhibiting the activation of JAK/PI3K/TRPV1 signaling pathway.
5.Forensic age assessment based on age-related changes in maxillofacial bones using CBCT
Shoutao NI ; Di SHAN ; Jilong ZHENG ; Yang LI ; Chaopeng YANG ; Weibo CAO ; Guanglong HE
Chinese Journal of Forensic Medicine 2025;40(2):181-187,193
Objective To establish a forensic age inference method based on age-related changes in maxillofacial CBCT imaging of Han population in North China.Methods CBCT imaging data of Han people aged 7~50 years were collected from September 2021 to September 2023.Mimics 17.0 software was used to perform 3D reconstruction on 480 cases that met the inclusion criteria(420 cases in the experimental group,60 cases in the blind test group,with a 50%male-to-female ratio)and obtain measured values for 16 indicators.The data were then statistically analyzed using SPSS 27.0 to identify age-related significant indicators and establish regression equations.Results Regression equations based on age-related significant indicators were established for different genders and age intervals.The accuracy of the equations decreased as the age interval increased.The blind test accuracy rates of regression equations for males across different age groups ranged between 62.6%and 80.6%,while for females,they ranged between 55.3%and 76.6%.Conclusion This study enriches maxillofacial skeletal data for Chinese population and establishes multivariate age inference regression equations based on significant indicators of age-related changes in maxillofacial CBCT,providing a new reference basis and inference method for forensic age assessment.
6.Forensic age assessment based on age-related changes in maxillofacial bones using CBCT
Shoutao NI ; Di SHAN ; Jilong ZHENG ; Yang LI ; Chaopeng YANG ; Weibo CAO ; Guanglong HE
Chinese Journal of Forensic Medicine 2025;40(2):181-187,193
Objective To establish a forensic age inference method based on age-related changes in maxillofacial CBCT imaging of Han population in North China.Methods CBCT imaging data of Han people aged 7~50 years were collected from September 2021 to September 2023.Mimics 17.0 software was used to perform 3D reconstruction on 480 cases that met the inclusion criteria(420 cases in the experimental group,60 cases in the blind test group,with a 50%male-to-female ratio)and obtain measured values for 16 indicators.The data were then statistically analyzed using SPSS 27.0 to identify age-related significant indicators and establish regression equations.Results Regression equations based on age-related significant indicators were established for different genders and age intervals.The accuracy of the equations decreased as the age interval increased.The blind test accuracy rates of regression equations for males across different age groups ranged between 62.6%and 80.6%,while for females,they ranged between 55.3%and 76.6%.Conclusion This study enriches maxillofacial skeletal data for Chinese population and establishes multivariate age inference regression equations based on significant indicators of age-related changes in maxillofacial CBCT,providing a new reference basis and inference method for forensic age assessment.
7.Artemin promotes proliferation and invasion of malignant peripheral nerve sheath tumor cells through the PI3K/Akt pathway
Hongliang ZHANG ; Haotian LIU ; Junyang LIU ; Chao ZHANG ; Ting LI ; Zhichao LIAO ; Yancheng LIU ; Jingyu ZHANG ; Kai ZHU ; Shuang LI ; Jinwei LIU ; Jilong YANG
Chinese Journal of Oncology 2025;47(2):149-159
Objective:To investigate the expression of Artemin (ARTN) in malignant peripheral nerve sheath tumor (MPNST), its effect on the malignant behavior of MPNST cells, and its signaling pathway.Methods:Fifty-one MPNST paraffin embedded tissues through surgical resection at Tianjin Medical University Cancer Hospital from January 1995 to November 2011 were collected, the expression of the ARTN protein was detected by immunohistochemistry, and the relationship between the ARTN protein expression and the clinical pathological characteristics and prognosis were analyzed. In human MPNST cell lines ST-8814 (NF-1) and STS26T(sporadic), ARTN overexpression and low expression cell lines were constructed by transfecting ARTN overexpression plasmids and ARTN small interfering RNA (siRNA), respectively. The expression of ARTN mRNA was detected by real time quantitative polymerase chain reaction (RT-qPCR), the expression of the ARTN protein and Phosphoinositide 3-kinase(PI3K)/Akt signaling pathway related proteins were detected by Western blot. CCK-8 assay was used to detect cell proliferation ability, and cell invasion assay was used to detect cell invasion ability. The pathway proteins that interacted with ARTN were searched in the STRING database, and the functional pathways were clarified by Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis. The PI3K/Akt pathway specific inhibitor LY294002 was used to block the PI3K/Akt pathway of ST-8814 and STS26T cells to observe the changes in cell proliferation and invasion.Results:Among the 51 MPNST tissue specimens, 22 cases showed a high expression of the ARTN protein and 29 cases showed a low expression of the protein. Higher expressions of the ARTN protein was associated with larger tumor diameters and disease progression (recurrence or metastasis) (both P<0.05). The median disease-free survival (DFS) of patients with a low expression of the ARTN protein was 26.2 months, and the median overall survival (OS) was 66.9 months. The median DFS and median OS of patients with a high expression of the ARTN protein were 10.7 months and 53.8 months, respectively. The log rank test results showed that the progression free survival rate of patients with a high expression of the ARTN protein was worse than that of patients with a low expression ( P=0.027), but the difference in overall survival rate between the two groups was not statistically significant ( P=0.790), which was also confirmed by Cox regression analysis. The CCK-8 assay results showed that after 48 hours of transfection, the absorbance ( A) values of ST-8814 and STS26T cells in the ARTN overexpression group were 1.35±0.01 and 1.10±0.02, respectively, which were higher than those in the empty plasmid control group (1.05±0.01 and 0.78±0.01, both P<0.01), while the A values of ST-8814 and STS26T cells in the ARTN siRNA group were 0.35±0.01 and 0.61±0.01, respectively, which were lower than those in the control siRNA group (0.74±0.01 and 1.10±0.04, both P<0.01). The results of cell invasion assay showed that the number of transmembrane cells in ST-8814 and STS26T cells overexpressing ARTN was (29.67±2.08) and (31.67±2.08), respectively, which were higher than those in the empty plasmid control group [(20.00±1.00) and (24.33±1.15), both P<0.01]. The number of transmembrane cells in ST-8814 and STS26T cells in the ARTN siRNA group were (14.00±2.00) and (19.33±1.53), respectively, which were lower than those in the control siRNA group [(19.33±2.52) and (23.33±0.58), both P<0.05].The KEGG results showed that ARTN is associated with multiple tumor signaling pathways, especially the PI3K/Akt signaling pathway. Western blot results showed that overexpression of ARTN upregulated the expression of p-PI3K and p-Akt proteins in ST-8814 and STS26T cells (both P<0.01).After knocking down ARTN expression, the expression of p-PI3K and p-Akt proteins was significantly down regulated (both P<0.01). LY294002 could significantly inhibit the effect of ARTN overexpression on ST-8814 and STS26T cells after blocking the PI3K/Akt pathway. The A values of ST-8814 and STS26T cells in the ARTN overexpression+LY294002 group were 1.09±0.06 and 0.82±0.01, respectively, which were lower than those in the ARTN overexpression group (1.50±0.01 and 1.29±0.01, respectively, both P<0.01). The numbers of transmembrane cells in the cell invasion assay were 16.67±3.21 and 19.67±2.31, respectively, which were also lower than those in the ARTN overexpression group (29.67±2.08 and 31.67±2.08, respectively, both P<0.01). Conclusions:In MPNST, a high expression of the ARTN protein was associated with larger tumor size, disease progression, and worse DFS. ARTN promotes the proliferation and invasion of MPNST cells through the PI3K/Akt signaling pathway.
8.Safety and efficacy of donor-derived chimeric antigen receptor T-cell therapy in patients with relapsed B-cell acute lymphoblastic leukemia after allogeneic hematopoietic stem cell transplantation
Yaqi ZHUO ; Sanfang TU ; Xuan ZHOU ; Jilong YANG ; Lijuan ZHOU ; Rui HUANG ; Yuxian HUANG ; Meifang LI ; Bo JIN ; Bo WANG ; Shiqi LI ; Zhongtao YUAN ; Lihua ZHANG ; Lin LIU ; Sanbin WANG ; Yuhua LI
Chinese Journal of Hematology 2024;45(1):74-81
Objective:To investigated the safety and efficacy of donor-derived CD19+ or sequential CD19+ CD22+ chimeric antigen receptor T-cell (CAR-T) therapy in patients with B-cell acute lymphoblastic leukemia (B-ALL) after allogeneic hematopoietic stem cell transplantation (allo-HSCT).Methods:The data of 22 patients with B-ALL who relapsed after allo-HSCT and who underwent donor-derived CAR-T therapy at the Zhujiang Hospital of Southern Medical University and the 920th Hospital of Joint Logistics Support Force of the People’s Liberation Army of China from September 2015 to December 2022 were retrospectively analyzed. The primary endpoint was overall survival (OS), and the secondary endpoints were event-free survival (EFS), complete remission (CR) rate, and Grade 3-4 adverse events.Results:A total of 81.82% ( n=18) of the 22 patients achieved minimal residual disease-negative CR after CAR-T infusion. The median follow-up time was 1037 (95% CI 546–1509) days, and the median OS and EFS were 287 (95% CI 132-441) days and 212 (95% CI 120-303) days, respectively. The 6-month OS and EFS rates were 67.90% (95% CI 48.30%-84.50%) and 58.70% (95% CI 37.92%-79.48%), respectively, and the 1-year OS and EFS rates were 41.10% (95% CI 19.15%-63.05%) and 34.30% (95% CI 13.92%-54.68%), respectively. Grade 1-2 cytokine release syndrome occurred in 36.36% ( n=8) of the patients, and grade 3-4 occurred in 13.64% of the patients ( n=3). Grade 2 and 4 graft-versus-host disease occurred in two patients. Conclusion:Donor-derived CAR-T therapy is safe and effective in patients with relapsed B-ALL after allo-HSCT.
9.Prevalence of Echinococcus infections in small rodents in Yushu City, Qinghai Province in 2023
Gengcheng HE ; Shusheng WU ; Xianglan QIN ; Jilong MA ; Tingjun YU ; Chengxi SONG ; Xiaojin MO ; Xiao MA ; Jianfeng BA ; Guirong ZHENG ; Bin JIANG ; Tian TIAN ; Shijie YANG ; Ting ZHANG ; Xiaonong ZHOU
Chinese Journal of Schistosomiasis Control 2024;36(2):169-173
Objective To investigate the prevalence of Echinococcus infections in small rodents around human residential areas in Yushu City, Qinghai Province in 2023, so as to provide insights into precision echinococcosis control. Methods One or two quadrats, each measuring 50 m × 50 m, were randomly assigned in Shanglaxiu Township and Longbao Township, Yushu City, Qinghai Province on June 2023, respectively, and 300 plate-type mouse traps, each measuring 12.0 cm × 6.5 cm, were assigned in each quadrat. Small rodents were captured during the period between 10 : 00 and 18 : 00 each day for 4 days. Then, all captured small rodents were identified and dissected, and liver specimens with suspected Echinococcus infections were subjected to pathological examinations. The Echinococcus cytochrome c oxidase 1 (cox1) gene was amplified using PCR assay, and the sequence of the amplified product was aligned to that was recorded in the GenBank to characterize the parasite species. In addition, a phylogenetic tree of Echinococcus was generated based on the cox1 gene sequence using the neighbor-joining method. Results A total of 236 small rodents were captured in Shanglaxiu and Longbao townships, Yushu City, including 65 Qinghai voles and 51 plateau pikas in Shanglaxiu Township, and 62 Qinghai voles and 58 plateau pikas in Longbao Township, and there was no significant difference in the constituent ratio of small rodents between the two townships (χ2 = 0.294, P > 0.05). Seven plateau pikas and 12 Qinghai voles were suspected to be infected with Echinococcus by dissection, and pathological examinations showed unclear structure of hepatic lobules and disordered hepatocyte arrangement in livers of small rodents suspected of Echinococcus infections. PCR assay identified E. shiquicus DNA in 7 Qinghai voles, which were all captured from Shanglaxiu Township. Phylogenetic analysis showed that the cox1 gene sequence of Echinococcus in small rodents was highly homologous to the E. shiquicus cox1 gene sequence reported previously. Conclusion Plateau pika and Qinghai vole were predominant small rodents around human residential areas in Yushu City, Qinghai Province in 2023, and E. shiquicus infection was detected in Qinghai voles.
10.Definition and Extraction of Traditional Chinese Medicine Inspection Gait Features Based on Three-Dimensional Key Points of the Human Body
Aihua GUAN ; Jilong SHEN ; Ziyan WANG ; Qi ZHANG ; Tao YANG ; Xufeng LANG ; Jiadong XIE ; Kongfa HU
Journal of Nanjing University of Traditional Chinese Medicine 2024;40(12):1331-1339
OBJECTIVE To analyze the differences in gait features between patients with cardiovascular and cerebrovascular dis-eases and normal people,and to explore new objective features of traditional Chinese medicine(TCM)whole-body inspection.METHODS A monocular camera was used to collect frontal walking videos of subjects,and the diagnosis results of TCM practitioners were used as disease annotation data;a deep learning model was used to estimate the three-dimensional coordinates of key points;the gait features were defined and calculated based on the three-dimensional coordinates of key points of the lower limbs;differences in gait features among people with cardiovascular and cerebrovascular diseases were collected and verified.RESULTS The three-di-mensional coordinates of key points of the lower limbs were automatically extracted and 8 types of TCM gait features were calculated:step width,stride length,foot lift height,limb angle,left and right hip joint angles,and left and right knee joint angles.It was found that there were significant differences in the features between people with cardiovascular and cerebrovascular diseases and healthy peo-ple(P<0.05).CONCLUSION The TCM inspection gait extracted by this study can effectively distinguish patients with cardiovas-cular and cerebrovascular diseases from healthy people,expands the research scope of TCM whole-body inspection,and provides new ideas for the early detection and prevention of cardiovascular and cerebrovascular diseases.

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