1.TSPO deficiency exacerbates acute lung injury via NLRP3 inflammasome-mediated pyroptosis
Jingyi HAN ; Xue ZHANG ; Menghua CAI ; Feng TIAN ; Yi XU ; Hui CHEN ; Wei HE ; Jianmin ZHANG ; Hui TIAN
Chinese Medical Journal 2024;137(13):1592-1602
Background::Acute respiratory distress syndrome (ARDS) is a common cause of respiratory failure in many critically ill patients. Although inflammasome activation plays an important role in the induction of acute lung injury (ALI) and ARDS, the regulatory mechanism of this process is still unclear. When cells are stimulated by inflammation, the integrity and physiological function of mitochondria play a crucial part in pyroptosis. However, the underlying mechanisms and function of mitochondrial proteins in the process of pyroptosis are largely not yet known. Here, we identified the 18-kDa translocator protein (TSPO), a mitochondrial outer membrane protein, as an important mediator regulating nucleotide-binding domain, leucine-rich repeat, and pyrin domain-containing protein 3 (NLRP3) inflammasome activation in macrophages during ALI.Methods::TSPO gene knockout (KO) and lipopolysaccharide (LPS)-induced ALI/ARDS mouse models were employed to investigate the biological role of TSPO in the pathogenesis of ARDS. Murine macrophages were used to further characterize the effect of TSPO on the NLRP3 inflammasome pathway. Activation of NLRP3 inflammasome was preformed through LPS + adenosine triphosphate (ATP) co-stimulation, followed by detection of mitochondrial membrane potential, reactive oxygen species (ROS) production, and cell death to evaluate the potential biological function of TSPO. Comparisons between two groups were performed with a two-sided unpaired t-test. Results::TSPO-KO mice exhibited more severe pulmonary inflammation in response to LPS-induced ALI. TSPO deficiency resulted in enhanced activation of the NLRP3 inflammasome pathway, promoting more proinflammatory cytokine production of macrophages in LPS-injured lung tissue, including interleukin (IL)-1β, IL-18, and macrophage inflammatory protein (MIP)-2. Mitochondria in TSPO-KO macrophages tended to depolarize in response to cellular stress. The increased production of mitochondrial damage-associated molecular pattern led to enhanced mitochondrial membrane depolarization and pyroptosis in TSPO-KO cells. Conclusion::TSPO may be the key regulator of cellular pyroptosis, and it plays a vital protective role in ARDS occurrence and development.
2.Accuracy of bedside lung ultrasound in predicting postoperative pulmonary complications in patients undergoing radical resection of gastrointestinal cancer
Jianmin JING ; Weiwei ZHANG ; Zhiqiang FANG ; Nirong WANG ; Yuehong QI ; Yan CHENG ; Jiaqi ZHANG ; Ying XUE ; Shuzhen YU
Chinese Journal of Anesthesiology 2023;43(8):937-941
Objective:To evaluate the accuracy of bedside lung ultrasound in predicting postoperative pulmonary complications (PPCs) in the patients undergoing radical resection of gastrointestinal cancer.Methods:One hundred and eight patients of both sexes, aged >18 yr, undergoing elective radical resection of gastrointestinal cancer with general anesthesia, were enrolled in the study. Lung ultrasound was performed before surgery (T 1) and at 2, 4 and 7 days after surgery (T 2-4). Lung ultrasound score (LUS) and B-line score were recorded. Serum procalcitonin (PCT) concentrations and blood routine were recorded, and systemic immune-inflammatory index (SII) was calculated. All the patients underwent chest CT examination before surgery and 7 days after surgery. The results of chest CT and clinical diagnosis were used as the gold standard for PPCs. The occurrence of PPCs within 7 days after surgery was recorded. The patients were divided into PPCs group and non-PPCs group according to the development of PPCs. Spearman′s correlation analysis was used to analyze the correlation of B-line score and LUS with PPCs, PCT and SII. The receiver operating curve was used to evaluate the accuracy of B-line score and LUB in predicting PPCs. Results:One hundred and three patients were finally enrolled in the study, including 45 patients in PPCs group and 58 patients in non-PPCs group, and the incidence of PPCs was 43.7%. Both B-line score and LUS were positively correlated with PPCs at T 1 ( P<0.001), and B-line score and LUS were positively correlated with PCT and SII at T 2-4 ( P<0.001). The AUC (95% confidence interval) of B-line score and LUB in predicting PPCs were 0.926 (0.879-0.972) and 0.909 (0.852-0.965), respectively ( P<0.001), the best cut-off values of B-line score and LUB in predicting PPCs were set at 25.5 and 11.5 respectively, and the sensitivity and specificity of B-line score were 0.80 and 0.88 respectively, and the sensitivity and specificity of LUB were 0.78 and 0.93 respectively. Conclusions:Bedside pulmonary ultrasonography (B-line score and LUS) can accurately predict the occurrence of PPCs in the patients undergoing radical resection of gastrointestinal cancer and dynamically evaluate the condition of PPCs, and B-line score >25.5 and LUS score >11.5 indicate a high risk of PPCs.
3.Study on the etiological characteristics and prevention and control of adult community-acquired pneumonia in hospitalized patients in a hospital in Beijing from 2015 to 2019
Mei WANG ; Jianyu ZHAO ; Xue LI ; Liyuan WU ; Qianqian ZHOU ; Yanfei HUANG ; Wenjun SUI ; Shaoya ZHANG ; Jie XU ; Jianmin JIN ; Haitong GU ; Xinxin LU
Chinese Journal of Preventive Medicine 2021;55(12):1410-1418
Objective:To explore the distribution characteristics of pathogens in adult patients with community-acquired pneumonia (CAP) and to provide basis for the diagnosis, treatment, prevention of CAP.Methods:1 446 inpatients with CAP were prospectively enrolled in a third-class hospital in Beijing in recent 5 years (from January 2015 to December 2019). Respiratory tract samples were collected for smear, culture, nucleic acid, antigen and antibody detection to identify the pathogen of CAP. Mann-Whitney U test was used for continuous variables and χ 2 test or Fisher′s exact test was used for categorical data for statistical analysis. Results:Among the 1 446 patients, 822 (56.85%) patients were infected with a single pathogen, 231 (15.98%) patients were infected with multiple pathogens, and 393 (27.18%) patients were not clear about the pathogen. Influenza virus is the first pathogen of CAP (20.95%, 303/1 446), mainly H1N1 (8.51%, 123/1 446), followed by mycoplasma pneumoniae (7.19%, 104/1 446), Mycobacterium tuberculosis (5.33%, 77/1 446) and Streptococcus pneumoniae (5.05%, 73/1 446). The outbreak of H1N1 occurred from December 2018 to February 2019, and the epidemic of mycoplasma pneumoniae pneumonia was monitored from August to November 2019. Patients under 65 years old had high detection rates of Mycoplasma pneumoniae (14.41% vs. 2.41%, χ2=74.712, P<0.001), Streptococcus pneumoniae (8.16% vs. 2.99%, χ2=18.156, P<0.001), rhinovirus (6.08% vs. 3.56%, χ2=5.025, P<0.025), Chlamydia pneumoniae (5.90% vs. 1.15%, χ2=26.542, P<0.001) and adenovirus (3.13% vs. 0.92%, χ2=9.547, P=0.002). The severe disease rate of CAP was 14.66% (212/1 446), and the average mortality rate was 3.66% (53/1 446). The severe illness rate and mortality rate of bacterial-viral co-infection were 28.97% (31/107) and 19.63% (21/107), respectively. Conclusions:Influenza virus is the primary pathogen of adult CAP. Outbreaks of Mycoplasma pneumoniae and H1N1 were detected in 2018 and 2019, respectively. The remission rate and mortality rate of virus-bacteria co-infection were significantly higher than those of single pathogen infection. Accurate etiological basis not only plays a role in clinical diagnosis and treatment, but also provides important data support for prevention and early warning.
4.Inconsistent and consistent classifications for small liver lesions by contrast-enhanced ultrasound LI-RADS and CECT/MRI LI-RADS
Xue LIANG ; Jinbin PAN ; Jianmin DING ; Fengmei WANG ; Lei LONG ; Yan ZHOU ; Yandong WANG ; Xiang JING
Chinese Journal of Ultrasonography 2021;30(11):938-943
Objective:To explore the inconsistent and consistent classifications for lesions ≤2 cm by contrast-enhanced ultrasound(CEUS) Liver Imaging Reporting and Data System(LI-RADS) v2017 and contrast-enhanced computed tomography/contrast-enhanced magnetic resonance imaging(CECT/MRI) LI-RADS v2018.Methods:The focal liver lesions ≤2 cm underwent CEUS and CECT/MRI within 1 month were enrolled in this retrospective study.Each nodule was categorized according to the CEUS LI-RADS v2017 and CECT/MRI LI-RADS v2018. Intermodality agreement between the CEUS LI-RADS and CECT/MRI LI-RADS for each lesion was assessed with Cohen′s Kappa. Lesions with inconsistent classification for CEUS LI-RADS and CECT/MRI were analyzed.Results:A total of 145 lesions with a size of (1.65±0.33)cm in 145 patients were included. The numbers of lesions in LR-3, 4, 5 and M were 16, 23, 90 and 16 on CEUS LI-RADS, 25, 31, 87 and 2 on CECT/MRI, respectively. And 73.1% lesions were classified as LR-5 or M on CEUS, while 61.4% lesions were classified as LR-5 or M on CECT/MRI ( P=0.033). The incidences of HCC in LR-3, 4 and 5 were 37.5%, 52.2% and 97.8% on CEUS LI-RADS, 56.0%, 64.5% and 96.6% on CECT/MRI LI-RADS respectively. Among the 145 lesions, 56 lesions had inconsistent classifications of CEUS and CECT/MRI LI-RADS. Twenty-eight lesions in CECT/MRI LR-3 and 4 were escalated to LR-4 and 5 by CEUS and 82.1% of them were found to be HCC. Fourteen lesions on CEUS LR-3 and 4 were escalated to LR-4 and 5 by CECT/MRI and 85.7% of them were found to be HCC. Conclusions:The LR-5 of the CEUS and CECT/EOB-MRI LI-RADS has a comparable incidence of HCC. However, the inter-modality agreement of the LI-RADS category between CEUS and CECT/EOB-MRI is poor. The proportion of lesions in CEUS LR-5 and M is much higher than that in CECT/MRI LR-5 and M, while the proportion of lesions in CECT/MRI LR-3 and 4 is high than that in CEUS LR-3 and 4.
5.Expression of serum Cav-1 and YKL-40 in acute cerebral infarction and the value of combined detection in prognosis evaluation
Jinyan WANG ; Liping JIAO ; Lisha HAO ; Jianmin ZHOU ; Qian XUE ; Aixia SONG
Journal of Chinese Physician 2021;23(2):231-235
Objective:To explore the expression of caveolin-1 (Cav-1) and Chitinase-40 (YKL-40) in acute cerebral infarction and the value of combined detection in prognosis evaluation.Methods:118 patients with acute cerebral infarction admitted to the First Affiliated Hospital of Hebei Northern University from January 2016 to June 2019 were selected as the research objects. According to the cerebral infarction volume, the patients were divided into small infarction group (<5 cm 3), middle infarction group (5-10 cm 3) and large infarction group (>10 cm 3). 108 healthy people were selected as the healthy control group. The serum levels of Cav-1 and YKL-40 were compared in the 3 groups, and the correlation between the degree of cerebral infarction and serum levels of Cav-1 and YKL-40 was analyzed. Receiver operating characteristic curve (ROC) was used to analyze the diagnostic value of the expression levels of Cav-1 and YKL-40 in patients with acute cerebral infarction; the patients were followed up for one year and the prognosis was evaluated by modified Rankin Scale (mRS); the correlation between serum Cav-1 and YKL-40 and prognosis was analyzed. Results:The expression levels of serum Cav-1 and YKL-40 in patients with acute cerebral infarction were significantly higher than those in healthy group ( P<0.001). The serum levels of Cav-1 and YKL-40 were positively correlated with the infarct volume of acute cerebral infarction ( r=0.854, P=0.004; r=0.867, P=0.002). ROC curve analysis showed that the sensitivity, Youden index and area under ROC curve of Cav-1 (21.78 μg/L) combined with YKL-40 (158.69 ng/ml) in the diagnosis of acute cerebral infarction were 85.59%, 0.532 and 0.896 (95% CI: 0.741-0.932), respectively, which were significantly higher than those of single index ( P<0.05). At 8 and 12 months of follow-up, the proportion of death and mRS score in the positive group were significantly higher than those in the negative group ( P<0.05). Conclusions:The serum Cav-1 and YKL-40 levels are significantly higher in patients with acute cerebral infarction. The combined examination of Cav-1 and YKL-40 can improve the diagnostic efficiency and has potential application value for early diagnosis and prognosis prediction of patients.
6.Study on the etiological characteristics and prevention and control of adult community-acquired pneumonia in hospitalized patients in a hospital in Beijing from 2015 to 2019
Mei WANG ; Jianyu ZHAO ; Xue LI ; Liyuan WU ; Qianqian ZHOU ; Yanfei HUANG ; Wenjun SUI ; Shaoya ZHANG ; Jie XU ; Jianmin JIN ; Haitong GU ; Xinxin LU
Chinese Journal of Preventive Medicine 2021;55(12):1410-1418
Objective:To explore the distribution characteristics of pathogens in adult patients with community-acquired pneumonia (CAP) and to provide basis for the diagnosis, treatment, prevention of CAP.Methods:1 446 inpatients with CAP were prospectively enrolled in a third-class hospital in Beijing in recent 5 years (from January 2015 to December 2019). Respiratory tract samples were collected for smear, culture, nucleic acid, antigen and antibody detection to identify the pathogen of CAP. Mann-Whitney U test was used for continuous variables and χ 2 test or Fisher′s exact test was used for categorical data for statistical analysis. Results:Among the 1 446 patients, 822 (56.85%) patients were infected with a single pathogen, 231 (15.98%) patients were infected with multiple pathogens, and 393 (27.18%) patients were not clear about the pathogen. Influenza virus is the first pathogen of CAP (20.95%, 303/1 446), mainly H1N1 (8.51%, 123/1 446), followed by mycoplasma pneumoniae (7.19%, 104/1 446), Mycobacterium tuberculosis (5.33%, 77/1 446) and Streptococcus pneumoniae (5.05%, 73/1 446). The outbreak of H1N1 occurred from December 2018 to February 2019, and the epidemic of mycoplasma pneumoniae pneumonia was monitored from August to November 2019. Patients under 65 years old had high detection rates of Mycoplasma pneumoniae (14.41% vs. 2.41%, χ2=74.712, P<0.001), Streptococcus pneumoniae (8.16% vs. 2.99%, χ2=18.156, P<0.001), rhinovirus (6.08% vs. 3.56%, χ2=5.025, P<0.025), Chlamydia pneumoniae (5.90% vs. 1.15%, χ2=26.542, P<0.001) and adenovirus (3.13% vs. 0.92%, χ2=9.547, P=0.002). The severe disease rate of CAP was 14.66% (212/1 446), and the average mortality rate was 3.66% (53/1 446). The severe illness rate and mortality rate of bacterial-viral co-infection were 28.97% (31/107) and 19.63% (21/107), respectively. Conclusions:Influenza virus is the primary pathogen of adult CAP. Outbreaks of Mycoplasma pneumoniae and H1N1 were detected in 2018 and 2019, respectively. The remission rate and mortality rate of virus-bacteria co-infection were significantly higher than those of single pathogen infection. Accurate etiological basis not only plays a role in clinical diagnosis and treatment, but also provides important data support for prevention and early warning.
7.DPHL:A DIA Pan-human Protein Mass Spectrometry Library for Robust Biomarker Discovery
Zhu TIANSHENG ; Zhu YI ; Xuan YUE ; Gao HUANHUAN ; Cai XUE ; Piersma R. SANDER ; Pham V. THANG ; Schelfhorst TIM ; Haas R.G.D. RICHARD ; Bijnsdorp V. IRENE ; Sun RUI ; Yue LIANG ; Ruan GUAN ; Zhang QIUSHI ; Hu MO ; Zhou YUE ; Winan J. Van Houdt ; Tessa Y.S. Le Large ; Cloos JACQUELINE ; Wojtuszkiewicz ANNA ; Koppers-Lalic DANIJELA ; B(o)ttger FRANZISKA ; Scheepbouwer CHANTAL ; Brakenhoff H. RUUD ; Geert J.L.H. van Leenders ; Ijzermans N.M. JAN ; Martens W.M. JOHN ; Steenbergen D.M. RENSKE ; Grieken C. NICOLE ; Selvarajan SATHIYAMOORTHY ; Mantoo SANGEETA ; Lee S. SZE ; Yeow J.Y. SERENE ; Alkaff M.F. SYED ; Xiang NAN ; Sun YAOTING ; Yi XIAO ; Dai SHAOZHENG ; Liu WEI ; Lu TIAN ; Wu ZHICHENG ; Liang XIAO ; Wang MAN ; Shao YINGKUAN ; Zheng XI ; Xu KAILUN ; Yang QIN ; Meng YIFAN ; Lu CONG ; Zhu JIANG ; Zheng JIN'E ; Wang BO ; Lou SAI ; Dai YIBEI ; Xu CHAO ; Yu CHENHUAN ; Ying HUAZHONG ; Lim K. TONY ; Wu JIANMIN ; Gao XIAOFEI ; Luan ZHONGZHI ; Teng XIAODONG ; Wu PENG ; Huang SHI'ANG ; Tao ZHIHUA ; Iyer G. NARAYANAN ; Zhou SHUIGENG ; Shao WENGUANG ; Lam HENRY ; Ma DING ; Ji JIAFU ; Kon L. OI ; Zheng SHU ; Aebersold RUEDI ; Jimenez R. CONNIE ; Guo TIANNAN
Genomics, Proteomics & Bioinformatics 2020;18(2):104-119
To address the increasing need for detecting and validating protein biomarkers in clinical specimens, mass spectrometry (MS)-based targeted proteomic techniques, including the selected reaction monitoring (SRM), parallel reaction monitoring (PRM), and massively parallel data-independent acquisition (DIA), have been developed. For optimal performance, they require the fragment ion spectra of targeted peptides as prior knowledge. In this report, we describe a MS pipe-line and spectral resource to support targeted proteomics studies for human tissue samples. To build the spectral resource, we integrated common open-source MS computational tools to assemble a freely accessible computational workflow based on Docker. We then applied the workflow to gen-erate DPHL, a comprehensive DIA pan-human library, from 1096 data-dependent acquisition (DDA) MS raw files for 16 types of cancer samples. This extensive spectral resource was then applied to a proteomic study of 17 prostate cancer (PCa) patients. Thereafter, PRM validation was applied to a larger study of 57 PCa patients and the differential expression of three proteins in prostate tumor was validated. As a second application, the DPHL spectral resource was applied to a study consisting of plasma samples from 19 diffuse large B cell lymphoma (DLBCL) patients and 18 healthy control subjects. Differentially expressed proteins between DLBCL patients and healthy control subjects were detected by DIA-MS and confirmed by PRM. These data demonstrate that the DPHL supports DIA and PRM MS pipelines for robust protein biomarker discovery. DPHL is freely accessible at https://www.iprox.org/page/project.html?id=IPX0001400000.
8."J. D" technique: A method for in situ fenestration of left subclavian artery in thoracic endovascular aortic repair
LIU Jidong ; YANG Wengang ; GU Jianmin ; XUE Song
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2020;27(04):450-453
Objective To report a simple and safe method for in situ fenestration of left subclavian artery in thoracic endovascular aortic repair (TEVAR). Methods Twenty-eight patients received in situ fenestration of left subclavian artery in TEVAR from June 2018 to May 2019 in our center, including 23 males and 5 females at an average age of 57.7±9.6 years. Among them, 12 patients used adjustable sheath or guiding catheter (a group A) and 16 patients used "J. D" technique (a group B). The clinical efficacy of the two groups was compared. Results In the group A, 1 patient failed to receive fenestration and was transferred to the chimney technique. In the group B, 1 patient due to the traction system shift during operation, was completed by traditional adjustable sheath puncture. The group B had shorter alignment-perforation time and trigger time and less complications. There was no significant difference in endoleak during short-term follow-up between the two groups. Conclusion The "J. D" technique is simple, safe and easy to obtain materials. It effectively reduces the risk caused by difficult sheath alignment during the in situ fenestration of the left subclavian artery. Although the results of recent follow-up are not significantly different from traditional methods, it still needs to accumulate the cases to observe the possible risks and difficulties.
9.Aucubin regulates skin damage repair by TGF-β/P38MAPK signaling pathway
Liu YANG ; Jianmin LI ; Yeqiu WANG ; Lifeng AN ; Jingwen HUANG ; Hui XUE ; Lihong ZHANG
International Journal of Traditional Chinese Medicine 2019;41(6):603-607
Objective To explore the mechanism of transforming growth factor-beta/p38 mitogen-activated protein kinases (TGF-β/P38MAPK) signaling pathway that transformed corallin into a transforming growth factor-beta/p38 mitogen activated protein kinases (TGF-β/P38MAPK).Methods The hair follicle stem cells (HFSCs) were divided into blank group,positive group and 10-10,10-9,10-8,10-7,10-6,10-5,10-4,10-3 mol/L mycoralin solution groups according to the random number table method.After 24 h,the cell proliferation rate was determined by MTT assay.The HFSCs were divided into blank group,positive group and low,medium and high dose groups according to the random number table method.The positive group was added with 5 mol/L of alkaline fibroblast growth factor solution,while the low,medium and high dose groups were added with 10-7,10-6 and 10-5 mol/L of mycoralin solution for intervention.The cell proliferation rate was determined by MTT assay.The protein expressions ofTGF-β,phosphorylated P38(p-p38),P38,Bax and bcl-2 in cells were detected by Western blot,and mRNA expressions of Bax and bcl-2 in cells of each group were detected by real-time quantitative PCR.Results Compared with the blank group,the productivity of the cells (0.418 ± 0.031,0.412 ± 0.014,0.468 ± 0.024 vs.0.353 ± 0.006) in the 10-7,10-6 and 10-5 mol/L aucubin groups significantly increased (P<0.01).Compared with the blank group,the protein expression of TGF-β (0.377 ± 0.027,0.338 ± 0.021,0.322 ± 0.017 vs.0.557 ± 0.017),p-p38 (0.270 ± 0.020,0.228 ± 0.013,0.216 ± 0.012 vs.0.461 ± 0.012),Bax (0.450 ± 0.017,0.365 ± 0.011,0.279 ± 0.006 vs.0.551 ± 0.015) in the low,medium and high dose groups significantly down-regulated,while the Bcl-2 (0.450 ± 0.017,0.365 ± 0.011,0.279 ± 0.006 vs.0.358 ± 0.011) protein expression significantly increased.Compared with the blank group,the expression of Bcl-2 (1.714 ± 0.028,2.514 ± 0.054,3.382 ± 0.084 vs.1.000 ± 0) mRNA increased,Bax (0.415 ± 0.020,0.353 ± 0.090,0.235 ± 0.114 vs.1.000 ± 0) mRNA in the low,medium and high dose groups significantly down-regulated (P<0.01).Conclusions By regulating the TGF-β/p38MAPK signaling pathway,mycoralin can reduce the expression of downstream apoptotic factors,and promote the repair of damaged skin.
10.Study on Quality Standard Improvement of Pheretima
Shasha WANG ; Yue QU ; Daquan XUE ; Lanqing LI ; Yang XIANG ; Baohui ZHANG
China Pharmacy 2019;30(17):2379-2383
OBJECTIVE: To provide reference for improving the quality standard of Pheretima. METHODS: The contents of hypoxanthine and inosine in medicinal material samples were determined by HPLC. HPLC fingerprint of Pheretima was established according to “Similarity evaluation system for TCM chromatogramtic fingerprint” (2012 edition) software, and similarity evaluation was conducted. The determination was performed on Purospher STAR RP-18 endcapped with mobile phase consisted of methanol-water (gradient elution) at the flow rate of 1 mL/min. The detection wavelength was 248 nm, and the column temperature was set at 30 ℃. The sample size was 20 μL. RESULTS: The results of methodological investigation of content determination showed that the linear range of hypoxanthine and inosine were 1.58-31.6 μg/mL (r=0.999 9), 5.52-110.4 μg/mL(r=0.999 8), respectively. limits of quantify were 0.316, 0.552 μg/mL, respectively; limits of detection were 0.158, 0.110 μg/mL, respectively; RSDs of precision, stability (24 h) and repeatability tests were all less than 2.0% (n=6). Average recovery rates were 103.0% (RSD=1.7%, n=6) and 101.2% (RSD=1.2%, n=6), respectively. HPLC fingerprint for 15 batches of samples were established, and 8 common peaks were identified. The similarity of HPLC fingerprint of 14 batches of sample with control fingerprint R was higher than 0.900. CONCLUSIONS: The established method for content determination of hypoxanthine and inosine and HPLC fingerprint of Pheretima are simple, accurate and reproducible, and can be used for quality control of Pheretima.

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