1.Efficacy Mechanism of Xianlian Jiedu Prescription Against Colorectal Cancer Recurrence vias Regulating Angiogenesis
Yanru XU ; Lihuiping TAO ; Jingyang QIAN ; Weixing SHEN ; Jiani TAN ; Chengtao YU ; Minmin FAN ; Changliang XU ; Yueyang LAI ; Liu LI ; Dongdong SUN ; Haibo CHENG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(6):79-87
ObjectiveTo explore effect of Xianlian Jiedu prescription on the recurrence of colorectal cancer (CRC) and investigate the related mechanisms. MethodsA postoperative recurrence model was established in 25 Balb/c mice by injecting CT26 cells subcutaneously into the armpit, followed by surgical removal of 99% of the subcutaneous tumor. The mice were randomly divided into model group, low-dose Xianlian Jiedu prescription (XLJDP-L) group (6.45 g·kg-1·d-1), medium-dose Xianlian Jiedu prescription (XLJDP-M) group (12.9 g·kg-1·d-1), high-dose Xianlian Jiedu prescription (XLJDP-H) group (25.8 g·kg-1·d-1), and 5-fluorouracil (5-FU) group (1×10-3 g·kg-1·d-1). The mice were euthanized after 14 days of continuous intervention, and recurrent tumor tissue was harvested. Hematoxylin and eosin (HE) staining was used to observe pathological and morphological changes in the recurrent tumor tissue. Immunohistochemistry (IHC) was employed to assess the expression of proliferating cell nuclear antigen (Ki67), vascular endothelial growth factor (VEGF), and platelet-endothelial cell adhesion molecule (CD31) in recurrent tumor tissue. The Western blot was used to detect the protein expression levels of angiopoietin-2 (ANG-2), VEGF, phosphorylated-protein kinase B (p-Akt), protein kinase B (Akt), phosphorylated-phosphatidylinositol 3-kinase (p-PI3K), and phosphatidylinositol 3-kinase (PI3K) in recurrent tumor tissue. ResultsBefore treatment, there were no statistical differences in tumor volume, tumor weight, and body mass among the XLJDP-L, XLJDP-M, and XLJDP-H groups and the 5-FU group compared to the model group, indicating model stability. After treatment, compared with those in the model group, the tumor volume and tumor weight in the XLJDP-L, XLJDP-M, and XLJDP-H groups and the 5-FU group were significantly reduced (P<0.01), showing dose dependency. Meanwhile, there were no significant differences in body weight among the XLJDP-L, XLJDP-M, and XLJDP-H groups and the 5-FU group compared to the model group. HE staining showed that compared with that in the model group, tumor tissue in the XLJDP-L, XLJDP-M, and XLJDP-H groups and the 5-FU group had loosely arranged cells, increased intercellular spaces, small and shriveled nuclei, light staining, fewer mitotic figures and atypical nuclei, and increased necrotic areas. IHC showed that compared with those of the model group, the positive rates of Ki67, VEGF, and CD31 in the recurrent tumor tissue of the XLJDP-L, XLJDP-M, and XLJDP-H groups and the 5-FU group were significantly reduced (P<0.01) in a dose-dependent manner. Western blot results showed that compared with those of the model group, the protein expression levels of ANG-2 and VEGF in the recurrent tumor tissue of the XLJDP-L, XLJDP-M, and XLJDP-H groups and the 5-FU group were significantly downregulated (P<0.05, P<0.01), and the p-Akt/Akt and p-PI3K/PI3K ratios were significantly decreased in a dose-dependent manner (P<0.05, P<0.01). ConclusionXianlian Jiedu prescription significantly inhibits the recurrence of CRC in mice after subcutaneous tumor surgery. The mechanism may involve regulating the PI3K/Akt pathway and downregulating key angiogenic proteins such as ANG-2, VEGF, and CD31.
2.Single-cell RNA sequencing reveals Shen-Bai-Jie-Du decoction retards colorectal tumorigenesis by regulating the TMEM131-TNF signaling pathway-mediated differentiation of immunosuppressive dendritic cells.
Yuquan TAO ; Yinuo MA ; Limei GU ; Ye ZHANG ; Qinchang ZHANG ; Lisha ZHOU ; Jie PAN ; Meng SHEN ; Xuefei ZHUANG ; Linmei PAN ; Weixing SHEN ; Chengtao YU ; Dan DONG ; Dong ZHANG ; Tingsheng LING ; Yang SUN ; Haibo CHENG
Acta Pharmaceutica Sinica B 2025;15(7):3545-3560
Colorectal tumorigenesis generally progresses from adenoma to adenocarcinoma, accompanied by dynamic changes in the tumor microenvironment (TME). A randomized controlled trial has confirmed the efficacy and safety of Shen-Bai-Jie-Du decoction (SBJDD) in preventing colorectal tumorigenesis. However, the mechanism remains unclear. In this study, we employed single-cell RNA sequencing (scRNA-seq) to investigate the dynamic evolution of the TME and validated cell infiltration with multiplex immunohistochemistry and flow cytometry. Bulk RNA sequencing was utilized to assess the underlying mechanisms. Our results constructed the mutually verifiable single-cell transcriptomic atlases in Apc Min/+ mice and clinical patients. There was a marked accumulation of CCL22+ dendritic cells (DCs) and an enhanced immunosuppressive action, which SBJDD and berberine reversed. Combined treatment with cholesterol and lipopolysaccharide induced characteristic gene expression of CCL22+ DCs, which may represent "exhausted DCs". Intraperitoneal injection of these DCs after SBJDD treatment eliminated its therapeutic effects. TMEM131 derived CCL22+ DCs generation by TNF signaling pathway and may be a potential target of berberine in retarding colorectal tumorigenesis. These findings emphasize the role of exhausted DCs and the regulatory mechanisms of SBJDD and berberine in colorectal cancer (CRC), suggesting that the multi-component properties of SBJDD may help restore TME homeostasis and offer novel cancer therapy.
3.Analysis of feasibility of selectively preserving the first branch of the right gastro-omental artery using bidirectional dissection in laparoscopic pylorus-preserving gastrectomy
Zeyao YE ; Pengfei YU ; Yang CAO ; Tengjiao CHAI ; Binzhong ZHANG ; Jun SIMA ; Bing WANG ; Zhihui JIANG ; Pingyuan YU ; Weixing WU ; Yi'an DU
Chinese Journal of Gastrointestinal Surgery 2024;27(8):840-845
Objective:To assess the safety and feasibility of selectively preserving the first branch of the right gastro-omental artery using bidirectional dissection in laparoscopic pylorus-preserving gastrectomy (LPPG).Methods:In this retrospective analysis, we studied preoperative, intraoperative, postoperative, and follow-up data of 30 patients with early gastric cancer treated in the Department of Gastric Cancer in Zhejiang Cancer Hospital (28 patients), Department of Gastrointestinal Surgery in Jiaxing Second Hospital (one patient) and Department of Gastrointestinal Surgery in Hangzhou Red Cross Hospital (one patient) who had undergone selective preservation of the first branch of the right gastro-omental artery during LPPG. The main variables studied were as follows: (1) intraoperative preservation of the first branch of the right gastro-omental artery; (2) the overall surgical situation; and (3) postoperative small bowel follow-through and endoscopy findings.Results:LPPG with selective preservation of the right gastro-omental artery vascular branch was achieved in all 30 of the study patients. The mean operation time was (244.3±29.3) minutes and the median intraoperative blood loss 50 (20–200) mL. The median tumor diameter was 1.2 (0.5–3.6) cm and an average of 32.3±11.6 lymph nodes were dissected. The overall median number of positive lymph nodes was 0 (0–6), and of No. 6 lymph nodes 5.1±1.5. Postoperative feeding resumed at an average of 5.2±0.5 days and the postoperative hospital stay averaged 8.4±3.4 days. Pathological stages were as follows: T1a (14 cases), T1b (10 cases), and T2 (6 cases). Small bowel follow-through imaging showed good results in 28 patients 5 days post-surgery, the remaining two exhibiting good results 9 days post-surgery. There were no instances of delayed gastric emptying, and only one patient (3.3%) developed intra-abdominal infection (resolved with conservative treatment).Conclusion:Selective preservation of the right gastro-omental artery during laparoscopic early gastric cancer surgery is a safe and feasible procedure for treating early mid-gastric body cancer with pyloric preservation.
4.Analysis of feasibility of selectively preserving the first branch of the right gastro-omental artery using bidirectional dissection in laparoscopic pylorus-preserving gastrectomy
Zeyao YE ; Pengfei YU ; Yang CAO ; Tengjiao CHAI ; Binzhong ZHANG ; Jun SIMA ; Bing WANG ; Zhihui JIANG ; Pingyuan YU ; Weixing WU ; Yi'an DU
Chinese Journal of Gastrointestinal Surgery 2024;27(8):840-845
Objective:To assess the safety and feasibility of selectively preserving the first branch of the right gastro-omental artery using bidirectional dissection in laparoscopic pylorus-preserving gastrectomy (LPPG).Methods:In this retrospective analysis, we studied preoperative, intraoperative, postoperative, and follow-up data of 30 patients with early gastric cancer treated in the Department of Gastric Cancer in Zhejiang Cancer Hospital (28 patients), Department of Gastrointestinal Surgery in Jiaxing Second Hospital (one patient) and Department of Gastrointestinal Surgery in Hangzhou Red Cross Hospital (one patient) who had undergone selective preservation of the first branch of the right gastro-omental artery during LPPG. The main variables studied were as follows: (1) intraoperative preservation of the first branch of the right gastro-omental artery; (2) the overall surgical situation; and (3) postoperative small bowel follow-through and endoscopy findings.Results:LPPG with selective preservation of the right gastro-omental artery vascular branch was achieved in all 30 of the study patients. The mean operation time was (244.3±29.3) minutes and the median intraoperative blood loss 50 (20–200) mL. The median tumor diameter was 1.2 (0.5–3.6) cm and an average of 32.3±11.6 lymph nodes were dissected. The overall median number of positive lymph nodes was 0 (0–6), and of No. 6 lymph nodes 5.1±1.5. Postoperative feeding resumed at an average of 5.2±0.5 days and the postoperative hospital stay averaged 8.4±3.4 days. Pathological stages were as follows: T1a (14 cases), T1b (10 cases), and T2 (6 cases). Small bowel follow-through imaging showed good results in 28 patients 5 days post-surgery, the remaining two exhibiting good results 9 days post-surgery. There were no instances of delayed gastric emptying, and only one patient (3.3%) developed intra-abdominal infection (resolved with conservative treatment).Conclusion:Selective preservation of the right gastro-omental artery during laparoscopic early gastric cancer surgery is a safe and feasible procedure for treating early mid-gastric body cancer with pyloric preservation.
5.Fire needle therapy combined with bladder function training for neurogenic bladder caused by spinal cord injury: a randomized controlled trial.
Yan DONG ; Zhengang LIU ; Yuan LIU ; Huarong LI ; Ran YU ; Weixing LIU ; Xiurong YANG ; Dongsheng WANG
Chinese Acupuncture & Moxibustion 2024;44(12):1395-1400
OBJECTIVE:
To observe the clinical effect and safety of fire needle therapy combined with bladder function training on neurogenic bladder (NB) caused by spinal cord injury.
METHODS:
A total of 60 patients with NB caused by spinal cord injury were randomly divided into an observation group and a control group , with 30 cases in each group. On the basis of conventional treatment with western medicine, the bladder function training was adopted in the control group, once a day and for 4 weeks. In the observation group, on the basis of the interventions as the control group, fire needling was operated at bilateral Sanyinjiao (SP 6) and Guanyuan (CV 4) and Zhongji (CV 3), once every two days and for 4 weeks (14 interventions in total). Separately, at the baseline and in 2 and 4 weeks of interventions, the urination conditions (average daily urination frequency, average daily leakage frequency, average daily single urination volume) were recorded in the two groups; the urodynamic parameters (maximum flow rate [Qmax], maximum detrusor pressure at maximum flow rate [PdetQmax], residual urine volume [RUA], maximum cystometric capacity [MCC], and bladder pressure) were detected; the neurogenic bladder symptom score (NBSS), urinary symptom distress score (USDS) were observed. Before and after treatment, the score of World Health Organization quality of life assessment scale-brief (WHOQOL-BREF) was observed in the two groups. The therapeutic effect, the incidence of urinary infection, and the safety were evaluated.
RESULTS:
In 2 and 4 weeks of interventions, the average daily urination frequency, the average daily leakage frequency, RUA, and the scores of NBSS and USDS decreased in the two groups when compared with the baseline (P<0.05). In 4 weeks of interventions, the above-mention outcomes were lower than those in 2 weeks of interventions (P<0.05); and the results in the observation group were lower in 2 and 4 weeks of interventions when compared with the control group (P<0.05). In 2 and 4 weeks of interventions, the average daily single urination volume, Qmax, PdetQmax, MCC, and bladder pressure increased in the two groups compared with the baseline (P<0.05). In 4 weeks of interventions, the above-mention outcomes were elevated in comparison with those in 2 weeks of interventions (P<0.05); and except for bladder pressure, the results in the observation group in 2 and 4 weeks of interventions were higher when compared with the control group (P<0.05). In 4 weeks of interventions, the scores of each dimension and the total scores of WHOQOL-BREF increased in comparison with the baseline in the two groups (P<0.05), and the scores of the observation group were higher than those of the control group (P<0.05). The incidence of urinary infection was 0% (0/30) in the observation group and 10.0% (3/30) in the control group, without significant difference (P>0.05). The total effective rate of the observation group was 93.3% (28/30), which was higher than that (73.3%, 22/30) of the control group (P<0.05). No serious adverse reactions occurred in the patients of the observation group.
CONCLUSION
Fire needle therapy combined with bladder function training can effectively relieve the clinical symptoms, ameliorate urination, restore bladder function and improve the quality of life in the patients with NB caused by spinal cord injury. This therapeutic regimen presents a high safety in practice.
Humans
;
Spinal Cord Injuries/therapy*
;
Female
;
Male
;
Acupuncture Therapy
;
Middle Aged
;
Adult
;
Urinary Bladder, Neurogenic/etiology*
;
Urinary Bladder/physiopathology*
;
Urination
;
Young Adult
;
Treatment Outcome
;
Aged
;
Adolescent
;
Combined Modality Therapy
6.Anesthesiologists′ proficiency and training needs in flexible bronchoscope-guided awake fiberoptic intubation in China: a nationwide prevalence survey
Dingding WANG ; Wei WEI ; Li WEI ; Lili FENG ; Hongjun LIU ; Yilei SHEN ; Junming XIA ; Weixing LI ; Yirong CAI ; Yuan HAN ; Huafeng WEI ; Wenxian LI ; Buwei YU
Chinese Journal of Anesthesiology 2023;43(7):832-835
Objective:To investigate the Chinese anesthesiologist′s proficiency, training experience and needs of flexible bronchoscope-guided awake flexible bronchoscopy intubation (AFBI) using a questionnaire method.Methods:The cluster sampling was used, and self-designed questionnaires that addressed 54 questions in 5 categories were distributed through WeChat and online platforms. The survey took one month, and the answers were automatically recorded by the WeChat server.Results:A total of 1 250 anesthesiologists participated in the survey in 30 provinces of China, 9 of them were not anesthesiologists, and 1 241 (99.28%) questionnaires were validated. In the valid questionnaires, 52.70% (654) of the anesthesiologists were from tertiary hospitals, and 74.78% (928) of the anesthesiologists were attending physicians or above, only 7.57% (94) of the anesthesiologists had sufficient confidence in AFBI. Twenty-five point two two percent (313) of the anesthesiologists preferred fiberoptic intubation as the first tool when dealing with the anticipated difficult airway. Forty-eight point one one percent (597) of the anesthesiologists had implemented AFBI. Among them, 80.74% (482) had experienced unsuccessful AFBI practices. Eight hundred and ninety-four anesthesiologists had received AFBI training, and the most common AFBI training strategy was theoretical lectures. In addition, the degree of satisfaction regarding the theoretical lectures quality, technical training, clinical practice relativity and non-technical skills training was 21.47% (192), 14.32% (128), 12.3% (110) and 17.90% (160), respectively. The degree of satisfaction with all the 4 training elements mentioned above was 7.27% (65).Conclusions:The awareness and practice of Chinese anesthesiologists in terms of clinical application of AFBI to treat difficult airways need to be strengthened at present, and the lack of high-quality AFBI training may be the key.
7.Comparison of Quality of Life of the Patients Three Months after Uniportal and Multiportal Thoracoscopic Lobectomy.
Qi ZHANG ; Wei DAI ; Xing WEI ; Run XIANG ; Hang GU ; Peihong HU ; Mingxin LIU ; Wei CHEN ; Huaijun GONG ; Yong LIANG ; Shichao ZHANG ; Weixing PENG ; Qiuling SHI ; Qiang LI ; Nanbin YU
Chinese Journal of Lung Cancer 2023;26(11):843-850
BACKGROUND:
The relationship between quality of life at three months after lung cancer surgery and different surgical approaches is remains unclear. This study aimed to compare the quality of life of patients three months after uniportal and multiportal thoracoscopic lobectomy.
METHODS:
Data from patients who underwent lung surgery at the Department of Thoracic Surgery, Sichuan Cancer Hospital between April 2021 and October 2021 were collected. The European Organization for Research and Treatment of Quality of Life Questionnaire-Core 30 (EORTC QLQ-C30) and Quality of Life Questionnaire-Lung Cancer 29 (EORTC QLQ-LC29) were used to collect quality of life data of the patients. Potential confounding factors in the baseline data were included in a multivariate regression model for adjustment, and the quality of life of the two groups three months postoperatively was compared with traditional clinical outcomes.
RESULTS:
A total of 130 lung cancer patients were included, with 57 males (43.8%) and 73 females (56.2%), and an average age of (57.1±9.5) yr. In the baseline data of the two groups, there was a statistical difference in the number of chest drainage tubes placed (P<0.001). After adjustment with the regression model, at three months postoperatively, there were no significant differences in all symptoms and functional status scores between the two groups (all P>0.05). The multiportal group had longer surgery time (120.0 min vs 85.0 min, P=0.001), postoperative hospital stay (6.0 d vs 4.0 d, P=0.020), and a higher incidence of early ≥ grade 2 complications (39.0% vs 10.1%, P=0.011) compared to the uniportal group.
CONCLUSIONS
Patients undergoing uniportal and multiportal thoracoscopic lobectomy have similar quality of life at three months postoperatively. The uniportal group may have certain advantages in terms of traditional clinical outcome indicators such as operation time, postoperative hospital stay, and early postoperative complications.
Male
;
Female
;
Humans
;
Lung Neoplasms/surgery*
;
Quality of Life
;
Thoracic Surgery, Video-Assisted/adverse effects*
;
Pneumonectomy/adverse effects*
;
Postoperative Complications/surgery*
;
Retrospective Studies
8.Efficacy and Mechanism of Shenbai Jiedu Prescription Against Proliferation of HCT116 Cells
Dong JIANG ; Haibo CHENG ; Weixing SHEN ; Changliang XU ; Jiani TAN ; Yueyang LAI ; Dongdong SUN ; Liu LI ; Minmin FAN ; Chengtao YU ; Jun XIAO
Chinese Journal of Experimental Traditional Medical Formulae 2022;28(13):34-41
ObjectiveTo investigate the mechanism by which Shenbai Jiedu prescription (SBJDF) inhibits the proliferation of colorectal cancer (CRC) HCT116 cells. MethodAfter 48 h treatment of HCT116 cells with SBJDF (0, 0.25, 0.5, 1, 2, 4 g·L-1), the viability of HCT116 cells were determined by methyl thiazolyl tetrazolium (MTT) colorimetry. Following the classification of cells into blank control group and SBJDF (1, 2, 4 g·L-1) groups, the effect of SBJDF on HCT116 cell morphology was observed under an inverted microscope. The effects of SBJDF on the proliferation of HCT116 cells and mitochondrial membrane potential (Δψm) were detected by colony formation assay and JC-1 probe, respectively. The flow cytometry was then performed for determining cell cycle distribution and apoptosis. The effects of SBJDF on cell cycle-, apoptosis-, and nuclear factor kappa-B (NF-κB) signaling pathway-related proteins were determined by Western blot. ResultSBJDF effectively inhibited the vitality of HCT116 cells and changed their morphology in a concentration-dependent manner. Compared with the blank control group, SBJDF at 1, 2, 4 g·L-1 significantly reduced cell colony formation (P<0.05, P<0.01),and SBJDF at 2 and 4 g·L-1 arrested the HCT116 cell cycle at G0/G1 phase (P<0.05, P<0.01). Compared with the blank control group, SBJDF at 1, 2, 4 g·L-1 remarkably down-regulated the protein expression of CyclinD1 (P<0.05, P<0.01). SBJDF at 2 and 4 g·L-1 lowered the CyclinA2 and cyclin-dependent kinase 4 (CDK4) (P<0.05, P<0.01). SBJDF at 4 g·L-1 reduced the cyclin-dependent kinase 1 (CDK1) (P<0.01). Compared with the blank control group, SBJDF at 2 and 4 g·L-1 induced HCT116 cell apoptosis, down-regulated the protein expression of anti-apoptosis-related proteins Bcl-2 and Bcl-xl as well as the NF-κB signaling pathway-related proteins IκB kinase α (IKKα),inhibitor α of NF-κB (IκBα),and phospho-NF-κB p65 (p-p65) (P<0.05, P<0.01), and diminished the mitochondrial membrane potential of HCT116 cells. ConclusionSBJDF inhibits the proliferation of HCT116 cells, which may be related to its inhibition of the activation of NF-κB signaling pathway and the induction of cell cycle arrest and apoptosis.
9.Shenbai Jiedu Prescription Inhibits Proliferation of Colorectal Cancer Cells by Regulating PTEN/PI3K/Akt Signaling Pathway
Jianrong LIU ; Min HUANG ; Minmin FAN ; Haibo CHENG ; Weixing SHEN ; Jun XIAO ; Changliang XU ; Jiani TAN ; Yueyang LAI ; Chengtao YU ; Dongdong SUN ; Liu LI
Chinese Journal of Experimental Traditional Medical Formulae 2022;28(14):36-43
ObjectiveTo study the mechanism of Shenbai Jiedu prescription inhibiting the proliferation of HCT116 colorectal cancer (CRC) cells by regulating the phosphatase and tensin homolog deleted on chromosome ten (PTEN)/phosphatidylinositol 3-kinase (PI3K)/ protein kinase B (Akt) signaling pathway. MethodShenbai Jiedu prescription was extracted by water extraction and alcohol precipitation to prepare freeze-dried powder. HCT116 cells were cultured in vitro, and treated with different concentrations of Shenbai Jiedu prescription (2, 4, 8, 16 g·L-1). The inhibitory effect of Shenbai Jiedu prescription on the proliferation of HCT116 cells was tested by methyl thiazolyl tetrazolium (MTT). Real-time quantitative PCR was used to detect the mRNA expression levels of PTEN, PI3K, Akt, glycogen synthase kinase-3β (GSK-3β), c-Myc, survivin and Cyclin D1. Western blot was employed to measure the protein expression levels of PTEN, phosphorylated PTEN (p-PTEN), PI3K, Akt, phosphorylated Akt (p-Akt), GSK-3β, phosphorylated GSK-3β (p-GSK-3β), c-Myc, survivin and Cyclin D1, β-catenin nuclear import was explored by immunofluorescence assay. ResultCompared with the control group, Shenbai Jiedu prescription inhibited the proliferation of HCT116 cells in a dose-dependent manner (P<0.01). Compared with the control group, the mRNA expression levels of PTEN and GSK-3β were up-regulated whereas those of PI3K, Akt, c-Myc, survivin and CyclinD1 were down-regulated after treatment with Shenbai Jiedu prescription (P<0.01). The protein expression levels of PTEN, p-PTEN and GSK-3β were up-regulated whereas those of PI3K, Akt, p-Akt, GSK-3β, p-GSK-3β, c-Myc, survivin and CyclinD1 were down-regulated (P<0.05, P<0.01). Immunofluorescence assay showed that Shenbai Jiedu prescription suppressed β-catenin nuclear import in HCT116 cells. ConclusionShenbai Jiedu prescription inhibited the proliferation of HCT116 cells via the mechanism of regulating the PTEN/PI3K/Akt signaling pathway.
10.Determination of potential genotoxic impurities chloroacetyl chloride and chloroacetic acid in azintamide raw material
Yu ZHOU ; Weixing NI ; Yong LIU ; Pengfei HUANG ; Mengxiang SU
Journal of China Pharmaceutical University 2022;53(3):300-305
In this paper, chemical derivatization-high performance liquid chromatography was used to determine the potential genotoxic impurities chloroacetyl chloride and chloroacetic acid, respectively, in the raw material of azintamide.Derivatization was carried out using 2-nitrophenylhydrazine followed by the determination.Separation was performed on a Thermo Syncronis C18 column (250 mm × 4.6 mm, 5 μm), with mobile phase consisting of 0.1% phosphoric acid in water (A) and acetonitrile(B) by gradient elution, at a flow rate of 1 mL/min.The column temperature was 40 °C and the detection wavelength was 226 nm.The blank solvent, derivatization reagent, and azintamide did not interfere with the peak of the test substance, and the target component was well separated from the others.For impurities chloroacetyl chloride and chloroacetic acid, the limits of detection (LOD) were 7.5 ng/mL and 15 ng/mL respectively. There was a good linear relationship between the integral area and the concentration in the range of 30-300 ng/mL.The sample recovery rate was in the range of 87.37% ~ 109.75%.The two methods established in this study have good specificity, good precision, high sensitivity and simple operation, which can be used for the trace determination of potential genotoxic impurities chloroacetyl chloride and chloroacetic acid in the raw material of azintamide.

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