1.Molecular Mechanisms of Salvia Miltiorrhiza and Its Active Ingredients against Colorectal Cancer: A Review
Jianing GUO ; Xiaochen NI ; Kaiyuan ZHANG ; Wei FAN ; Chuhang WANG ; Chao XU ; Jianbo HUANG ; Tao JIANG ; Guangji ZHANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(4):307-314
Colorectal cancer (CRC) is one of the most common cancers, with its incidence ranking high among cancers. It stands as the second leading cause of cancer-related death worldwide. In the early stages, CRC lacks specific symptoms, and most patients are diagnosed at advanced stages, making it a major research focus in the field of gastrointestinal tumors. Currently, clinical CRC treatments face several common challenges, including high surgical risks, frequent metastasis and recurrence, drug resistance, and significant side effects from chemotherapy and radiation therapy. With the development and application of traditional Chinese medicine (TCM), it has been found that TCM and its active ingredients can effectively inhibit CRC cell proliferation, invasion, migration, and angiogenesis, and promote apoptosis and autophagy, thereby slowing the progression of CRC. This has become a key focus of CRC treatment research. Salvia Miltiorrhiza has multiple pharmacological effects, including activating blood circulation to dispel blood stasis, unlocking meridians to relieve pain, clearing heat to calm irritability, and cooling blood to reduce abscesses. It contains a variety of chemical components, including diterpenoids, phenolic acids, flavonoids, polysaccharides, nitrogen-containing compounds, steroids, and lactone compounds. This review summarized the molecular mechanisms of Salvia miltiorrhiza and its active ingredients in the treatment of CRC. It is found that these ingredients exert anti-CRC effects through various molecular mechanisms, including cell cycle arrest, promotion of apoptosis, inhibition of cell invasion and migration, induction of autophagy, suppression of tumor angiogenesis, and remodeling of the tumor microenvironment. The review aims to provide new insights for the drug development and clinical application of Salvia miltiorrhiza in CRC treatment.
2.Effect of Ligustilide on Neutrophil Extracellular Traps in Rats with Cerebral Ischemia-reperfusion Injury
Qian WU ; Yang WANG ; Jianing ZHOU ; Zhihan WAN ; Ke HU ; Qi HUANG ; Ning WANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(8):82-88
ObjectiveTo explore the possible mechanisms by which ligustilide (LIG) exerts neuroprotective effects on ischemic stroke (IS) by inhibiting the release of neutrophil extracellular traps (NETs), promoting blood-brain barrier repair, and alleviating post-ischemic neuroinflammation, thereby providing a new direction for IS treatment. MethodsA middle cerebral artery occlusion (MCAO) model was established in rats. The rats were divided into the sham operation (Sham) group, model (Model) group, low- and high-dose LIG groups (20, 40 mg·kg-1), and the NET inhibitor CI-amidine group (CI-amidine, 10 mg·kg-1). Drug treatments were administered for 3 days. Neurological injury after ischemia was evaluated by 2,3,5-triphenyltetrazolium chloride (TTC) staining, neurological deficit scoring, and brain index measurement. Flow cytometry and Western blot were used to analyze changes in neutrophil expression. Immunofluorescence was used to observe the fluorescence intensity of the NET marker citrullinated histone H3 (H3Cit). Western blot was performed to detect the expression of blood-brain barrier tight junction-related proteins and inflammatory factors, including interleukin-18 (IL-18) and interleukin-1β (IL-1β). ResultsCompared with the Sham group, the Model group exhibited significant brain tissue injury (P<0.05), significantly increased neutrophil numbers and NET expression (P<0.05), significantly impaired blood-brain barrier permeability (P<0.05), and significantly increased expression of inflammatory factors (P<0.05). Compared with the Model group, both low- and high-dose LIG significantly alleviated brain tissue injury in rats (P<0.01), inhibited neutrophil numbers and NET expression (P<0.01), reduced blood-brain barrier damage (P<0.01), and suppressed the expression of inflammatory factors IL-18 and IL-1β (P<0.01), thereby ultimately exerting a neuroprotective effect. ConclusionThe neuroprotective effect of LIG in rats with cerebral ischemia-reperfusion injury may be related to inhibition of neutrophils and the NETs induced by them.
3.Effect of Ligustilide on Neutrophil Extracellular Traps in Rats with Cerebral Ischemia-reperfusion Injury
Qian WU ; Yang WANG ; Jianing ZHOU ; Zhihan WAN ; Ke HU ; Qi HUANG ; Ning WANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(8):82-88
ObjectiveTo explore the possible mechanisms by which ligustilide (LIG) exerts neuroprotective effects on ischemic stroke (IS) by inhibiting the release of neutrophil extracellular traps (NETs), promoting blood-brain barrier repair, and alleviating post-ischemic neuroinflammation, thereby providing a new direction for IS treatment. MethodsA middle cerebral artery occlusion (MCAO) model was established in rats. The rats were divided into the sham operation (Sham) group, model (Model) group, low- and high-dose LIG groups (20, 40 mg·kg-1), and the NET inhibitor CI-amidine group (CI-amidine, 10 mg·kg-1). Drug treatments were administered for 3 days. Neurological injury after ischemia was evaluated by 2,3,5-triphenyltetrazolium chloride (TTC) staining, neurological deficit scoring, and brain index measurement. Flow cytometry and Western blot were used to analyze changes in neutrophil expression. Immunofluorescence was used to observe the fluorescence intensity of the NET marker citrullinated histone H3 (H3Cit). Western blot was performed to detect the expression of blood-brain barrier tight junction-related proteins and inflammatory factors, including interleukin-18 (IL-18) and interleukin-1β (IL-1β). ResultsCompared with the Sham group, the Model group exhibited significant brain tissue injury (P<0.05), significantly increased neutrophil numbers and NET expression (P<0.05), significantly impaired blood-brain barrier permeability (P<0.05), and significantly increased expression of inflammatory factors (P<0.05). Compared with the Model group, both low- and high-dose LIG significantly alleviated brain tissue injury in rats (P<0.01), inhibited neutrophil numbers and NET expression (P<0.01), reduced blood-brain barrier damage (P<0.01), and suppressed the expression of inflammatory factors IL-18 and IL-1β (P<0.01), thereby ultimately exerting a neuroprotective effect. ConclusionThe neuroprotective effect of LIG in rats with cerebral ischemia-reperfusion injury may be related to inhibition of neutrophils and the NETs induced by them.
4.Construction of A Conceptual Framework for the Integration of Traditional Chinese and Western Medicine in Evolutionary Syndrome Differentiation and Treatment Across Full-cycle of Parkinson's Disease
Yu WANG ; Jianing MEI ; Hongping ZHAO ; Yunzhe TANG ; Zijun WEI ; Qinliang TAO ; Xueyi HAN ; Jiyuan HU ; Yunyun ZHANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(10):270-279
Parkinson's disease (PD) is a complex neurodegenerative disease involving multiple systems and neurotransmitters. Due to the high clinical heterogeneity of PD,it is urgent to establish a comprehensive and long-term traditional Chinese medicine (TCM) management model. In this paper,the conceptual framework of full-cycle management of PD is preliminarily constructed:based on the evolution of the pathophysiological mechanisms of protein deposition and neurotransmitter disorder in PD,the three-stage syndrome characteristics of the prodromal stage (predominant healthy Qi with subtle pathogenic factors),the early clinical stage (declining healthy Qi with growing pathogenic factors) and the middle and late stages (overwhelming pathogenic factors with deficient healthy Qi) are longitudinally described. Through the syndrome differentiation of visceral manifestations,the etiology and pathogenesis of PD motor and non-motor symptoms were comprehensively analyzed,while the matching treatment methods and prescriptions were inferred,and the modular scheme of the combining main symptoms,accompanying symptoms and secondary symptoms was proposed. The conceptual gap of TCM regarding motor complications ('variable syndrome') and PD-related hyperpyrexia syndrome ('critical syndrome') was explained. This framework reflects the characteristics of combination of disease and syndrome and overall constant motion,and provides new theories and research ideas for individualized and whole-process management of PD in TCM.
5.Construction of A Conceptual Framework for the Integration of Traditional Chinese and Western Medicine in Evolutionary Syndrome Differentiation and Treatment Across Full-cycle of Parkinson's Disease
Yu WANG ; Jianing MEI ; Hongping ZHAO ; Yunzhe TANG ; Zijun WEI ; Qinliang TAO ; Xueyi HAN ; Jiyuan HU ; Yunyun ZHANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(10):270-279
Parkinson's disease (PD) is a complex neurodegenerative disease involving multiple systems and neurotransmitters. Due to the high clinical heterogeneity of PD,it is urgent to establish a comprehensive and long-term traditional Chinese medicine (TCM) management model. In this paper,the conceptual framework of full-cycle management of PD is preliminarily constructed:based on the evolution of the pathophysiological mechanisms of protein deposition and neurotransmitter disorder in PD,the three-stage syndrome characteristics of the prodromal stage (predominant healthy Qi with subtle pathogenic factors),the early clinical stage (declining healthy Qi with growing pathogenic factors) and the middle and late stages (overwhelming pathogenic factors with deficient healthy Qi) are longitudinally described. Through the syndrome differentiation of visceral manifestations,the etiology and pathogenesis of PD motor and non-motor symptoms were comprehensively analyzed,while the matching treatment methods and prescriptions were inferred,and the modular scheme of the combining main symptoms,accompanying symptoms and secondary symptoms was proposed. The conceptual gap of TCM regarding motor complications ('variable syndrome') and PD-related hyperpyrexia syndrome ('critical syndrome') was explained. This framework reflects the characteristics of combination of disease and syndrome and overall constant motion,and provides new theories and research ideas for individualized and whole-process management of PD in TCM.
6.Expression of long non-coding RNA HOXC13-AS in head and neck squamous cell carcinoma and its effect on the proliferation and migration of tumor cells
Dan WANG ; Xiaoya MA ; Jianing ZHAO ; Ming ZHAO
Chinese Archives of Otolaryngology-Head and Neck Surgery 2025;32(9):551-556
OBJECTIVE To investigate the expression of long non-coding RNA(lncRNA)HOXC13-AS in head and neck squamous cell carcinoma tissues and its effects on tumor cell proliferation and migration in head and neck tumor cell lines.METHODS Clinical samples from 26 patients with laryngeal squamous cell carcinoma and 4 patients with hypopharyngeal squamous cell carcinoma who were treated surgically in the Second Hospital of Jilin University from March 2024 to December 2024 were collected as research subjects.Head and neck squamous cell carcinoma tissue specimens and corresponding adjacent tissue specimens were selected.The expression of HOXC13-AS in 30 cases of head and neck squamous cell carcinoma tissues and corresponding adjacent tissues was detected by real-time fluorescence quantitative polymerase chain reaction(qRT-PCR).Clinical data were collected to analyze the correlation between HOXC13-AS expression and various clinical pathological factors.siRNA technology was used to interfere with the expression of HOXC13-AS in laryngeal cancer cell line TU212 and hypopharyngeal squamous cell line FaDu.Cell Counting Kit-8(CCK-8)assay was used to detect cell proliferation ability,and cell scratch assay was used to detect cell migration ability.RESULTS The expression level of lncRNA HOXC13-AS in head and neck squamous cell carcinoma tissues(12.60±11.26)was significantly higher than that in corresponding adjacent tissues(1.40±0.61),with a statistically significant difference(t=5.485,P<0.001).The expression of HOXC13-AS in head and neck squamous cell carcinoma tissues was closely related to T stage(P=0.004),lymph node metastasis(P=0.006),and clinical stage(P=0.007)of patients.CCK-8 results showed that the absorbance values of TU212 cell line groups at 24,48,and 72 h were si-NC group(0.373±0.010,0.738±0.026,1.003±0.124),si-HOXC13-AS-1 group(0.365±0.015,0.686±0.019,0.935±0.028),and si-HOXC13-AS-2 group(0.364±0.024,0.700±0.026,0.943±0.053),with statistically significant differences(F24 h=0.484,F48 h=7.893,F72 h=1.345,P<0.01);the absorbance values of FaDu cell line groups at 24,48,and 72 h were si-NC group(0.727±0.054,0.834±0.072,1.224±0.127),si-HOXC13-AS-1 group(0.532±0.005,0.650±0.079,1.021±0.044),and si-HOXC13-AS-2 group(0.647±0.088,0.687±0.025,1.074±0.055),with statistically significant differences(F24 h=16.143,F48 h=14.259,F72 h=9.409,P<0.001).Cell scratch assay showed that the migration rates of TU212 cell line groups at 12 and 24 h were si-NC group(38.971%±3.824%,69.185%±0.469%),si-HOXC13-AS-1 group(18.182%±2.580%,33.378%±2.302%),and si-HOXC13-AS-2 group(25.017%±0.288%,48.413%±0.805%),with statistically significant differences(F12 h=47.295,F24 h=471.745,P<0.0001);The migration rates of FaDu cell lines in each group at 12 and 24 hours were as follows:si-NC group(39.067%±3.196%,58.222%±0.448%),si-HOXC13-AS-1 group(13.689%±0.132%,39.358%±3.985%),and si-HOXC13-AS-2 group(23.335%±0.680%,35.526%±0.758%),with statistically significant differences(F12 h=138.1,F24 h=101.749,P<0.0001).CONCLUSION HOXC13-AS is significantly upregulated in head and neck squamous cell carcinoma tissues and cells,and is associated with the clinical pathological characteristics of patients.Silencing HOXC13-AS can significantly inhibit the proliferation and migration of head and neck tumor cells.
7.Expression of MCM2 and RF4 genes in liver cancer based on bioinformatics and experimental verification
Jianing GUO ; Pengyu ZHANG ; Xingfen WANG ; Yanchun QU ; Xianglian ZHANG ; Hui LI
International Journal of Biomedical Engineering 2025;48(1):77-87
Objective:To screen differentially expressed genes (DEGs) associated with liver cancer by bioinformatics analysis method, and to investigate the mechanism of the minichromosome maintenance protein 2 ( MCM2) and replication factor C subunit 4 ( RFC4) genes in liver cancer in vitro. Methods:Gene expression profiling data of 80 and 36 hepatocellular carcinoma tissues and 307 and 13 cirrhotic tissues were obtained from the GSE25097 and GSE98620 datasets of the gene expression analysis (GEO) database, respectively. Gene expression profiling data of 374 liver cancer tissues and 50 normal liver tissues were downloaded from the cancer genome atlas (TCGA) database. Limma and DESeqs R software were used to process the gene expression profiling data, construct protein-protein interaction networks, and analysis the relevance of these genes to survival. Weighted gene co-expression network analysis was performed to screen out the core genes. Liver cancer SMMC7721 cells were transfected with MCM2 blank plasmid (MCM2 control group), MCM2 overexpression plasmid [MCM2 WT1 group, MCM2 WT2 (2-fold WT1) group], RFC4 blank plasmid (RFC4 control group), and RFC4 overexpression plasmid [RFC4 WT1 group, RFC4 WT2 (2-fold WT1) group], respectively. The expression of MCM2 and RFC4 in liver cancer cell lines and their transfection levels were detected by real-time fluorescence quantitative reverse transcription PCR and Western blotting. The effects of MCM2 and RFC4 on the proliferation of hepatocellular carcinoma cells were detected by MTT assay and cell cloning assay, respectively. The effects of MCM2 and RFC4 on the migration of liver cancer cells were determined by the scratch assay. The effects of MCM2 and RFC4 on liver cancer cell invasion were detected by Transwell assay.Results:By bioinformatic analysis, 9 HCC DEGs were selected, including ubiquitin conjugating enzyme E2 T ( UBE2T), aurora kinase A ( AURKA), targeting protein for Xklp2 ( TPX2), MCM2, RFC4, ribonucleoside-diphosphate reductase subunit M2 ( RRM2), serine peptidase inhibitory factor Kazal type 1 ( SPINK1), collagen type XV alpha 1 chain ( COL15A1) and C-C motif chemokine 25 ( CCL25). Among the six genes associated with clinical stages, the MCM2 and RFC4 genes were found to be strongly associated with prognosis in liver cancer. The relative protein expression of MCM2 and RFC4 in HepG2 cells (1.83±0.07, 1.44±0.09) and SMMC7721 cells (1.74±0.05, 1.43±0.08) was higher than that in MIHA cells (1.00±0.02, 1.00±0.03), and all the differences were statistically significant (all P<0.05). The relative gene expression of MCM2 and RFC4 in HepG2 cells (14.30±0.12, 5.10±0.18) and SMMC7721 cells (10.60±0.11, 7.60±0.07) was higher than that in MIHA cells (1.00±0.05, 1.00±0.03), and all the differences were statistically significant (all P<0.05). Compared with the MCM2 control group, the absorbance values [(0.28±0.01 and 0.21±0.01) vs 0.18±0.03], the number of clonal cells [(717±12 and 782±29) cells vs (389±17) cells], the percentage migration [(0.43±0.02 and 0.68±0.01) vs 0.15±0.06], and the number of cellular invasions [(933±21 and 821±11) cells vs (409±16) cells] were higher in the MCM2 WT1 and MCM2 WT2 groups, and the differences were all statistically significant (all P<0.05). Compared with the RFC4 control group, the absorbance values [(0.30±0.02 and 0.21±0.01) vs 0.17±0.02], the number of cloned cells [(571±11 and 728±9) cells vs (373±23) cells], the percentage migration [(0.75±0.11 and 0.67±0.04) vs 0.34±0.07], and the number of cell invasion [(835±26 and 818±18) cells vs (629±12) cells] were higher in the RFC4 WT1 and the RFC4 WT2 groups, and the differences were statistically significant (all P<0.05). Conclusions:MCM2 and R FC4 genes play a role in promoting tumorigenesis and growth in hepatocellular carcinoma.
8.Morphologic and functional effect of core training combined with respiratory training on multifidus and transversus abdominis in patients with lumbar disc herniation
Jianing SONG ; Xiaole LOU ; Huan LIU ; Xue HAN ; Lei XU ; Min WANG
Chinese Journal of Rehabilitation Theory and Practice 2025;31(1):107-116
ObjectiveTo explore the effect of respiratory training based on core stabilization training on lumbar disc herniation. MethodsFrom January, 2023 to October, 2024, 96 patients with lumbar disc herniation admitted to the First Affiliated Hospital of Bengbu Medical University were divided into control group (n = 32), core group (n = 32) and respiratory group (n = 32). All the groups underwent conventional rehabilitation therapy, with core stabilization training in the core group and respiratory training combined with core stabilization training in the respiratory group, additionally, for four weeks. Before and after training, the scores of Visual Analogue Scale, Japanese Orthopaedic Association (JOA) and Oswestry Dysfunction Index (ODI) were compared, the average electromyographic value (AEMG) and root mean square (RMS) value of the multifidus and transversus abdominis were detected by surface electromyography (sEMG); and the thickness of the multifidus and transversus abdominis were measured by musculoskeletal ultrasonography bilaterally. ResultsThe intra-group effect (F > 597.796, P < 0.001), inter-group effect (F > 16.535, P < 0.001) and interaction effect (F > 49.622, P < 0.001) were significant in the scores of VAS, JOA and ODI; which were better in the respiratory group than in the control group and the core group (P < 0.05), and were better in the core group than in the control group (P < 0.001). The intra-group effect (F > 7971.631, P < 0.001), inter-group effect (F > 177.760, P < 0.001) and interaction effect (F > 478.771, P < 0.001) were significant in the thickness of the transversus abdominis and multifidus; which were better in the respiratory group than in the control group and the core group (P < 0.001), and were better in the core group than in the control group (P < 0.001). The intra-group effect (F > 144303.007, P < 0.001), inter-group effect (F > 1495.458, P < 0.001) and interaction effect (F > 3121.361, P < 0.001) were significant in the RMS of the multifidus and transversus abdominis; which were better in the respiratory group than in the control group and the core group (P < 0.001), and were better in the core group than in the control group (P < 0.001). The intra-group effect (F > 1890.532, P < 0.001), inter-group effect (F > 607.132, P < 0.001) and interaction effect (F > 824.923, P < 0.001) were significant in the AEMG of the multifidus and transversus abdominis; which were better in the respiratory group than in the control group and core group (P < 0.001), and were better in the core group than in the control group (P < 0.001). ConclusionCore training combined with respiratory training can more effectively reduce pain and improve dysfunction by enhancing the strength and control of the core muscles, thus improving the quality of life of patients with lumbar disc herniation.
9.Analysis of visual field manifestations of non-arteritic anterior ischemic optic neuropathy
Yue LI ; Ying WANG ; Wenxin JIAO ; Jilu LIN ; Jianing WANG
International Eye Science 2025;25(4):671-675
AIM: To observe the manifestations and distribution patterns of visual field in non-arteritic anterior ischemic optic neuropathy(NAION).METHODS: Retrospective observational study. The investigation encompassed 183 patients(246 eyes)diagnosed with NAION who were evaluated at the Neuro-Ophthalmology/Acupuncture Department within the Eye Hospital, China Academy of Chinese Medical Sciences from June 2018 to December 2023. Recorded clinical data covered demographic details, incidence, disease duration, presence of systemic diseases, and histories of tobacco and alcohol use, along with best-corrected visual acuity(BCVA), visual field index(VFI), type of visual field defect, and thickness of the peripapillary retinal nerve fiber layer(pRNFL).RESULTS: A total of 183 patients(246 eyes)were enrolled. The cohort consisted of 101 males and 82 females; 120 exhibited unilateral symptoms, while 63 showed bilateral symptoms, with a mean age of 56.20±9.78 years(range 29-81 years). The types of visual field defects observed were varied: 90 eyes(36.6%)had diffuse loss, 63 eyes(25.6%)experienced inferior hemifield loss, 32 eyes(13.0%)displayed ring scotomas, 22 eyes(8.9%)had arcuate scotomas, 11 eyes(4.5%)presented with quadrant defects, 15 eyes(6.1%)had sectorial or wedge defects, and 13 eyes(5.3%)showed superior hemifield loss. The BCVA(LogMAR)and VFI of patients with diffuse visual field loss were poorer than patients with other types of visual defects(all P<0.05). There were statistically significant differences in visual field defects among patients of different genders and ages(all P<0.05). However, history of hypertension, diabetes, hyperlipidemia, sleep apnea and other systemic diseases, history of smoking and alcohol, and course of disease did not show specificity in the NAION visual field(all P>0.05).CONCLUSION:NAION manifests with a broad spectrum of visual field impairments across different genders, age, and levels of visual functionality. Extensive future research is necessary to identify additional reasons influencing the types of visual field damage in NAION.
10.Tongue squamous cell carcinoma-targeting Au-HN-1 nanosystem for CT imaging and photothermal therapy.
Ming HAO ; Xingchen LI ; Xinxin ZHANG ; Boqiang TAO ; He SHI ; Jianing WU ; Yuyang LI ; Xiang LI ; Shuangji LI ; Han WU ; Jingcheng XIANG ; Dongxu WANG ; Weiwei LIU ; Guoqing WANG
International Journal of Oral Science 2025;17(1):9-9
Tongue squamous cell carcinoma (TSCC) is a prevalent malignancy that afflicts the head and neck area and presents a high incidence of metastasis and invasion. Accurate diagnosis and effective treatment are essential for enhancing the quality of life and the survival rates of TSCC patients. The current treatment modalities for TSCC frequently suffer from a lack of specificity and efficacy. Nanoparticles with diagnostic and photothermal therapeutic properties may offer a new approach for the targeted therapy of TSCC. However, inadequate accumulation of photosensitizers at the tumor site diminishes the efficacy of photothermal therapy (PTT). This study modified gold nanodots (AuNDs) with the TSCC-targeting peptide HN-1 to improve the selectivity and therapeutic effects of PTT. The Au-HN-1 nanosystem effectively targeted the TSCC cells and was rapidly delivered to the tumor tissues compared to the AuNDs. The enhanced accumulation of photosensitizing agents at tumor sites achieved significant PTT effects in a mouse model of TSCC. Moreover, owing to its stable long-term fluorescence and high X-ray attenuation coefficient, the Au-HN-1 nanosystem can be used for fluorescence and computed tomography imaging of TSCC, rendering it useful for early tumor detection and accurate delineation of surgical margins. In conclusion, Au-HN-1 represents a promising nanomedicine for imaging-based diagnosis and targeted PTT of TSCC.
Tongue Neoplasms/diagnostic imaging*
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Carcinoma, Squamous Cell/diagnostic imaging*
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Animals
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Gold/chemistry*
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Mice
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Photothermal Therapy/methods*
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Tomography, X-Ray Computed
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Photosensitizing Agents
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Metal Nanoparticles
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Humans
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Cell Line, Tumor

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