1.Advances of Fundamental Research on Traditional Chinese Medicine in Regulation of Tumor-associated Macrophages for the Prevention and Treatment of Lung Cancer Metastasis
LIU SHIHUI ; LI JIAXUAN ; QUE ZUJUN ; YU PAN ; TIAN JIANHUI
Chinese Journal of Lung Cancer 2024;27(7):541-549
Lung cancer is the leading cause of cancer-related deaths worldwide,with metastasis being the primary cause of mortality in lung cancer patients,and its prevention and control efficacy remain limited.In recent years,immunothera-py has emerged as a promising direction for overcoming the bottleneck of metastasis.Macrophages,as essential components of innate immunity,participate in the entire process of tumor initiation and progression.Tumor-associated macrophages(TAMs)represent the most abundant immune population in the tumor microenvironment(TME),displaying both anti-tumor M1-like and pro-tumor M2-like phenotypes.The latter promotes tumor invasion and metastasis,angiogenesis,lymphangiogenesis,immune suppression,and reactivation of dormant disseminated tumor cells(DTCs),thereby facilitating tumor metastasis.In recent years,traditional Chinese medicine(TCM)has shown significant efficacy in inhibiting tumor metastasis and has been extensively validated.It exerts anti-tumor effects by reducing the recruitment of TAMs,inhibiting M2-like polarization,and modulating cytokines and proteins in the TME.This paper reviews the relationship between TAMs and lung cancer metastasis,elucidates the targets and mechanisms of TCM in regulating TAMs to prevent and treat lung cancer metastasis,aiming to pro-vide insights into lung cancer prevention and treatment.
2.Initial Construction of the Tumor Metastatic State Doctrine under the Perspective of Integration of Chinese and Western Medicine
Jianhui TIAN ; Bin LUO ; Zujun QUE ; Yun YANG ; Jialiang YAO ; Yan LI
Journal of Traditional Chinese Medicine 2024;65(20):2065-2069
Based on the concepts of "people-oriented" in traditional Chinese medicine and "tumor-suppression" in modern medicine, we have combed the studies on the spatial and temporal evolution of tumor metastasis and its biological characteristics in different perspectives, and initially proposed the theory of tumor metastasis from the perspective of the dynamic game between the tumor cells and the body's immune system under the theory of the integration of Chinese and Western medicine, that is, the formation of metastasis is the result of the dynamic evolution of the cancer cells and their surrounding environmental factors in the body over time and space. It is believed that the symptomatic manifestation of metastasis is systematic, the triggering factors of metastasis are constant, and the clinical outcome of metastasis is staged. Accordingly, it is proposed to understand the mechanism of metastasis from the perspective of spatial and temporal dynamics, to establish a clinical and pathological model for identifying metastasis, and to reveal the critical point of metastasis, so as to facilitate the change of the research on tumor metastasis from static to dynamic, and provide ideas for the formulation of metastasis prevention and treatment strategies, and the construction of a new system of metastasis prevention and treatment in the clinical tumor field.
3.Expression Levels of Plasma Exosomal lncNORAD in Bladder Cancer and Its Clinical Diagnostic Value
Yuting GAO ; Dong LI ; Zujun SUN
Chinese Journal of Laboratory Medicine 2024;47(8):872-878
Objective:To evaluate the diagnostic efficacy of plasma exosomal lncNORAD as a tumor marker for bladder cancer (BCa) by examining its expression levels in BCa patients and analyzing the correlation with clinical pathological characteristics.Methods:This retrospective study. 100 BCa patients diagnosed between October 1, 2021, and October 1, 2023, at Tongji Hospital, Tongji University were randomly selected as the patient group (76 males, 24 females, mean age 71±10 years). In the same period, 40 patients with benign urinary system diseases (benign disease group; 33 males, 7 females, mean age 64±9 years) and 70 healthy individuals undergoing routine check-ups (control group; 53 males, 17 females, mean age 45±12 years) were included. Plasma exosomes were extracted using an exosome extraction kit, and morphologically and molecularly were characterized by transmission electron microscopy, nanoparticle tracking analysis, and Western blotting. Total RNA from plasma exosomes was extracted, and the expression level of exosomal lncNORAD was detected by real-time quantitative polymerase chain reaction. In the BCa patient group, patients were divided into high expression and low expression groups based on the median relative expression level of lncNORAD. The χ2 test was used to compare the relationship between lncNORAD expression and clinical pathological characteristics, and the receiver operating characteristic (ROC) curve was used to evaluate its diagnostic efficacy for BCa.Results:The expression level of plasma exosomal lncNORAD in the patient group was significantly higher than that in the control group ( P<0.001) and the benign disease group ( P<0.001). High expression of lncNORAD was closely related to TNM stage ( P=0.001, χ2=10.187), lymph node metastasis ( P=0.003, χ2=8.575), tumor grade ( P=0.006, χ2=7.440), and nuclear matrix protein 22 (NMP22, χ2=5.844) ( P=0.016). ROC analysis showed that the area under the curve (AUC) for diagnosing BCa with plasma exosomal lncNORAD was 0.828 (95% CI 0.772-0.883). The diagnostic efficacy was further improved when exosomal lncNORAD was combined with the conventional marker NMP22 (AUC=0.866, 95% CI 0.815-0.916). Conclusion:The upregulation of lncNORAD in plasma exosomes of BCa patients is associated with multiple clinical pathological characteristics, indicating its potential as a diagnostic biomarker for BCa.
4.Analysis of Key Genes and Immune Infiltration Mechanism of Primary Sj?gren's Syndrome and Prediction of Targeted Traditional Chinese Medicine Based on Bioinformatics
Xinzhu YUAN ; Lingqin LI ; Huan DU ; Changwei LIN ; Zujun YUAN ; Yanjiang WANG ; Baofu WANG ; Xisheng XIE
World Science and Technology-Modernization of Traditional Chinese Medicine 2023;25(11):3592-3604
Objective Using bioinformatics methods to study the immune infiltration mechanism of Primary Sj?gren's syndrome(pSS)and to explore potential target Chinese medicines,which can provide new directions for the clinical treatment of pSS.Methods Gene expression profile microarray dataset of pSS was downloaded from the GEO database,differential genes were screened using R software,and gene ontology(GO)and gene pathway enrichment(KEGG)enrichment analysis was performed on these differential genes.Protein interaction network analysis of differential genes was performed by applying the STRING database,key genes were screened by using Cytoscape,and ELISA for the verification of key genes expression.Immune infiltration and correlation of immune cells in pSS were calculated by CIBERSORT inverse convolution method in 22.Finally,the herbal prediction of key target genes was performed by using the Coremine Medical database.Results A total of 232 differential genes were obtained,of which 207 were up-regulated and 25 were down-regulated.GO was mainly enriched in:leukocyte mediated immunity,lymphocyte mediated immunity,leukocyte cell-cell adhesion,etc;KEGG was mainly enriched in Hematopoietic cell lineage,Primary immunodeficiency,Intestinal immune network for IgA production,Phagosome,Leishmaniasis.Ten key genes were screened:PTPRC,CD19,LCP2,CCR5 and CD69 etc.The hub genes expression in the pSS is the same as that of GSE40611.Immune infiltration showed that memory B cells,T cells CD4 memory activated,and T cells CD4 na?ve were highly expressed in the pSS.Immune cell correlation analysis showed a positive correlation between initial Monocytes and T cells regulatory(Tregs),a positive correlation between Macrophages M1 and B cell na?ve,and a negative correlation between Plasma cells and T cells CD4 memory activated.COREMINE Medical predicted that Ginseng,Panax notoginseng,Tripterygium wilfordii,Burnet,Magnolia,and Strychni may treat pSS.Conclusion The development and progression of pSS are the results of the combined involvement of multiple genes and pathways.Memory B cells,T cells CD4 memory activated,and T cells CD4 na?ve may promote the development of pSS.The predicted Ginseng,Panax notoginseng,Tripterygium wilfordii,Burnet,Magnolia,Strychni may be used as target herbs for the potential treatment of pSS.
5.Distribution and transfusion efficacy of unexpected antibody
Rong ZHANG ; Kai KANG ; Jiang XIE ; Yuwei LIN ; Zujun LI ; Jiping XU ; Zhengqiu LIAN ; Ling LI
Chinese Journal of Blood Transfusion 2023;36(12):1132-1135
【Objective】 To investigate the clinical characteristics and antibody distribution as well as evaluate the transfusion efficacy in unexpected antibody positive patients. 【Methods】 A total of 12 235 patients from January 1, 2022 to March 31, 2023 who hospitalized in our hospital and applied for blood transfusion were selected, and those with unexpected antibody were included. The clinical data, including gender, age, diagnosis, blood type, history of transfusion and pregnancy were collected for antibody distribution analysis. Patients who received transfusion were grouped according to the DAT results and the components of red blood cells transfused, and the Hb values of each group before and after transfusion were compared. 【Results】 Among12 235 patients, 118 were positive for antibody screening, with a prevalence of 0.96%. The antibodies from Rh system were the most common (27.43%, 48/175), followed by MNS system (8.57%, 15/175) and Lewis system (6.29%, 11/175), mainly anti-E (18.29%, 32/175), anti-M (8.00%, 14/175) and anti-Lea (5.71%, 10/175). In addition, 62 transfused patients were divided into group A with suspended red blood cell transfusion and group B with washed red blood cell transfusion for positive DAT, and group C for negative DAT. Hb values (g/L) pre- and post-transfusion were 59.19±15.67 vs 77.52±15.09 in group A, 56.35±14.08 vs 74.44±15.63 in group B, 56.00±12.06 vs 75.00±4.73 in group C, respectively. The Hb values of post-transfusion for three groups were all higher than those of pre-transfusion (P<0.05). 【Conclusion】 Anti-E from Rh system is the most common antibody in patients with unexpected antibody. Appropriate red blood cells transfusion with Hb increases by an average of 6-7 g/L per 1 U of red blood cells indicating good transfusion efficacy. For positive DAT patients, transfusion of suspended red blood cell is feasible.
6.Haploidentical transplantation has a superior graft-versus-leukemia effect than HLA-matched sibling transplantation for Ph- high-risk B-cell acute lymphoblastic leukemia
Menglin FAN ; Yu WANG ; Ren LIN ; Tong LIN ; Fen HUANG ; Zhiping FAN ; Yajing XU ; Ting YANG ; Na XU ; Pengcheng SHI ; Danian NIE ; Dongjun LIN ; Zujun JIANG ; Shunqing WANG ; Jing SUN ; Xiaojun HUANG ; Qifa LIU ; Li XUAN
Chinese Medical Journal 2022;135(8):930-939
Background::Compared with human leukocyte antigen (HLA)-matched sibling donor (MSD) transplantation, it remains unclear whether haploidentical donor (HID) transplantation has a superior graft-versus-leukemia (GVL) effect for Philadelphia-negative (Ph-) high-risk B-cell acute lymphoblastic leukemia (B-ALL). This study aimed to compare the GVL effect between HID and MSD transplantation for Ph- high-risk B-ALL.Methods::This study population came from two prospective multicenter trials (NCT01883180, NCT02673008). Immunosuppressant withdrawal and prophylactic or pre-emptive donor lymphocyte infusion (DLI) were administered in patients without active graft-versus-host disease (GVHD) to prevent relapse. All patients with measurable residual disease (MRD) positivity posttransplantation (post-MRD+) or non-remission (NR) pre-transplantation received prophylactic/pre-emptive interventions. The primary endpoint was the incidence of post-MRD+.Results::A total of 335 patients with Ph- high-risk B-ALL were enrolled, including 145 and 190, respectively, in the HID and MSD groups. The 3-year cumulative incidence of post-MRD+ was 27.2% (95% confidence interval [CI]: 20.2%-34.7%) and 42.6% (35.5%-49.6%) in the HID and MSD groups (P = 0.003), respectively. A total of 156 patients received DLI, including 60 (41.4%) and 96 (50.5%), respectively, in the HID and MSD groups ( P= 0.096). The 3-year cumulative incidence of relapse was 18.6% (95% CI: 12.7%-25.4%) and 25.9% (19.9%-32.3%; P = 0.116) in the two groups, respectively. The 3-year overall survival (OS) was 67.4% (95% CI: 59.1%-74.4%) and 61.6% (54.2%-68.1%; P = 0.382), leukemia-free survival (LFS) was 63.4% (95% CI: 55.0%-70.7%) and 58.2% (50.8%-64.9%; P= 0.429), and GVHD-free/relapse-free survival (GRFS) was 51.7% (95% CI: 43.3%-59.5%) and 37.8% (30.9%-44.6%; P= 0.041), respectively, in the HID and MSD groups. Conclusion::HID transplantation has a lower incidence of post-MRD+ than MSD transplantation, suggesting that HID transplantation might have a superior GVL effect than MSD transplantation for Ph- high-risk B-ALL patients.Trial registration::ClinicalTrials.gov: NCT01883180, NCT02673008.
7.Study on the expression of p62 protein in patients with lung adenocarcinoma
Bingjie ZENG ; Ping JI ; Zujun SUN ; Junlu WU ; Anquan SHANG ; Wenqiang QUAN ; Dong LI
Chinese Journal of Preventive Medicine 2021;55(5):685-690
To explore the expression of p62 protein in lung adenocarcinoma (LUAD). In this study, a cross-sectional study was adopted. From December 2011 to May 2013, 60 patients with lung adenocarcinoma who were diagnosed and treated in Tongji Hospital of Tongji University, Shanghai were selected for paraffin embedding and tissue chip preparation, and immunohistochemistry (IHC) technology was used to detect the expression of p62 in lung adenocarcinoma patients′ cancer tissues and adjacent tissues, and analyze the relationship between p62 expression and the clinicopathological characteristics and survival prognosis of patients with lung adenocarcinoma; at the same time, 6 cases of lung adenocarcinoma were selected by random sampling cancer tissues and adjacent tissues were detected by Western Blot (WB) to detect p62 protein and analyzed by gray value. Preoperative examination specimens of inpatients with lung adenocarcinoma diagnosed from April 2018 to early October 2019, and plasma specimens of healthy subjects were collected, and enzyme linked immunosorbent assay (ELISA) was used to detect lung adenocarcinoma patients and healthy patients. The expression of p62 in the plasma of the subjects was statistically analyzed using SPSS 22.0 software. The results of IHC showed that the positive expression rate of p62 in cancer tissues was significantly higher than that in adjacent tissues, and the difference was statistically significant ( t=5.593, P<0.001). Similarly, WB results showed that the expression of p62 protein in cancer tissues was significantly higher than that in adjacent tissues. It is statistically relevant ( t=2.238, P=0.049). The expression of p62 was statistically correlated with tumor size, clinicopathological stage and lymph node metastasis in patients with lung adenocarcinoma (all P<0.05). The overall survival of patients with lung adenocarcinoma with high p62 expression was worse than that of patients with low p62 expression (95 %CI was 0.238-0.870, P=0.028), suggesting that the high expression of p62 is related to the poor prognosis of patients with lung adenocarcinoma. The level of p62 protein in the plasma of patients with lung adenocarcinoma was significantly higher than that in the healthy control group. The difference was statistically significant ( t=8.533, P<0.001). The area under the receiver operating characteristic curve was 0.835 (95 %CI was 0.779-0.891, P<0.001), which is significantly higher than CEA, CA125, CA153 and other single traditional indicators, and the combined detection of four indicators has the highest diagnostic efficiency. p62 was strongly expressed in cancer tissues and serum, which is related to the poor prognosis and overall survival rate of LUAD patients.
8.Study on the expression of p62 protein in patients with lung adenocarcinoma
Bingjie ZENG ; Ping JI ; Zujun SUN ; Junlu WU ; Anquan SHANG ; Wenqiang QUAN ; Dong LI
Chinese Journal of Preventive Medicine 2021;55(5):685-690
To explore the expression of p62 protein in lung adenocarcinoma (LUAD). In this study, a cross-sectional study was adopted. From December 2011 to May 2013, 60 patients with lung adenocarcinoma who were diagnosed and treated in Tongji Hospital of Tongji University, Shanghai were selected for paraffin embedding and tissue chip preparation, and immunohistochemistry (IHC) technology was used to detect the expression of p62 in lung adenocarcinoma patients′ cancer tissues and adjacent tissues, and analyze the relationship between p62 expression and the clinicopathological characteristics and survival prognosis of patients with lung adenocarcinoma; at the same time, 6 cases of lung adenocarcinoma were selected by random sampling cancer tissues and adjacent tissues were detected by Western Blot (WB) to detect p62 protein and analyzed by gray value. Preoperative examination specimens of inpatients with lung adenocarcinoma diagnosed from April 2018 to early October 2019, and plasma specimens of healthy subjects were collected, and enzyme linked immunosorbent assay (ELISA) was used to detect lung adenocarcinoma patients and healthy patients. The expression of p62 in the plasma of the subjects was statistically analyzed using SPSS 22.0 software. The results of IHC showed that the positive expression rate of p62 in cancer tissues was significantly higher than that in adjacent tissues, and the difference was statistically significant ( t=5.593, P<0.001). Similarly, WB results showed that the expression of p62 protein in cancer tissues was significantly higher than that in adjacent tissues. It is statistically relevant ( t=2.238, P=0.049). The expression of p62 was statistically correlated with tumor size, clinicopathological stage and lymph node metastasis in patients with lung adenocarcinoma (all P<0.05). The overall survival of patients with lung adenocarcinoma with high p62 expression was worse than that of patients with low p62 expression (95 %CI was 0.238-0.870, P=0.028), suggesting that the high expression of p62 is related to the poor prognosis of patients with lung adenocarcinoma. The level of p62 protein in the plasma of patients with lung adenocarcinoma was significantly higher than that in the healthy control group. The difference was statistically significant ( t=8.533, P<0.001). The area under the receiver operating characteristic curve was 0.835 (95 %CI was 0.779-0.891, P<0.001), which is significantly higher than CEA, CA125, CA153 and other single traditional indicators, and the combined detection of four indicators has the highest diagnostic efficiency. p62 was strongly expressed in cancer tissues and serum, which is related to the poor prognosis and overall survival rate of LUAD patients.
9.Establishment of HPLC Fingerprint ,Chemical Pattern Recognition Analysis and Content Determination of the Leaves of Toricellia angulata from Different Regions
Zhongyao HAN ; Jun XIANG ; Jianyu CHEN ; Yiyong SONG ; Shiwai LI ; Wenshuang TANG ; Zujun YE ; Linsu ZHANG ; Hao TIAN ; Wanle WANG
China Pharmacy 2021;32(10):1224-1229
OBJECTIVE:To provide reference for the quality control of the leaves of Toricellia angulata . METHODS :HPLC method was adopted. The determination was performed on Agela Promosil C 18 column with 0.2% phosphoric acid solution-acetonitrile(gradient elution )as mobile phase at the flow rate of 1.0 mL/min. The detection wavelength was set at 210 nm,and column temperature was 35 ℃. The sample size was 10 μL. HPLC fingerprint of 10 batches of the leaves of T. angulata was established and similarity evaluation was conducted by using Similarity Evaluation System of TCM Chromatographic Fingerprint(2004 edition). The chromatographic peak was identified by comparing with the chromatogram of reference substance. Cluster analysis ,PCA and PLS-DA were used to identify chemical patterns ,and the quality differential markers were screened. The contents of hyperoside and isoquercitrin were determined by the same HPLC. RESULTS :The similarities of HPLC fingerprint of 10 batches of the leaves of T. angulata with control fingerprint were 0.923-0.983. A total of 11 common peaks were identified ,and the peaks 4 and 5 were hyperoside and isoquercitrin ,respectively. Results of cluster analysis ,PCA and PLS-DA showed that 10 batches of leaves of T. angulata could be divided into two categories ,Y10 was clustered into one category ,and others were clustered into one category. PLS-DA analysis showed that 6 common peaks (peaks 4,3,10,2,6 and 11) with variable importance projection (VIP)greater than 1 were selected. Average contents of hyperoside and isoquercitrin in 10 batches of the leaves of T. angulata were 0.47-6.97,0.21-1.87 mg/g,respectively. CONCLUSIONS :Established HPLC fingerprint and the method for content determination are stable and reliable ,and can be used for the quality control of the leaves of T. angulata from different areas. Six quality differential markers including hyperoside in the leaves of T. angulata from different areas are qnyz202034) preliminarily screened.
10.Expression level and clinical application value of human serum exosomal miR-218-5p in colorectal cancer
Wenjing CHANG ; Anquan SHANG ; Dianyu YANG ; Jiale TIAN ; Zujun SUN ; Dong LI ; Wenqiang QUAN
Chinese Journal of Laboratory Medicine 2021;44(8):709-714
Objective:To explore the expression level of exosomal miR-218-5p in the serum of patients with colorectal cancer (CRC) and its correlation with the clinical pathological characteristics, and evaluate its diagnostic efficacy in CRC.Methods:A group of 78 patients with colorectal cancer diagnosed in Tongji Hospital affiliated to Tongji University from October 2016 to October 2018 were selected. Blood was collected before operation and serum was preserved. Forty cases of healthy people in the same period were selected as the control group. ExoQuick kit was used to extract serum exosomes. Transmission electron microscope, NTA and western blot were used to identify the morphology and molecular phenotype of exosomes. MiRNeasy kit was used to extract total RNA in serum exosomes. Real-time fluorescent quantitative PCR (RT-qPCR) was used to detect the expression level of serum exosomal miR-218-5p in each group. CRC patients were divided into high expression and low expression groups using the median relative expression level of miR-218-5p as the cut off value, and the four-square chi-square test (χ 2 test) was used to judge the relationship between miR-218-5p and clinicopathological features. The receiver operating characteristic (ROC) curve was used to evaluate the diagnostic efficacy of miR-218-5p in colorectal cancer. Results:The exosomes in serum were successfully extracted by kit method. The expression level of serum exosomal miR-218-5p in patients with colorectal cancer was significantly lower than that in normal healthy people [0.566(0.364, 0.850) vs 1.054(0.781, 1.709), P<0.001]. The low expression level was significantly better then the correlated with tumor size, TNM stage, lymph node metastasis and depth of invasion (all P<0.05). Receiver operating characteristic (ROC) analysis showed that the area under the curve (AUC) of serum exosomal miR-218-5p for the diagnosis of CRC was 0.827 (95% CI 0.754-0.900), which was significantly better than the conventional tumor marker carcinoembryonic antigen CEA (AUC =0.718, 95% CI 0.626-0.811) and carbohydrate antigen CA199 (AUC = 0.661, 95% CI 0.564-0.758). Conclusions:The down-regulation of miR-218-5p expression in serum exosomes of colorectal cancer patients is associated with a variety of adverse clinicopathological factors, which has the potential to become a diagnostic biomarker for colorectal cancer.

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