1.Analysis of chemical constituents and components absorbed into plasma of Ardisia crenata based on UPLC-QE-HF-MS/MS
Hui SHI ; Xiao LI ; Ying ZHOU ; Jingxin DING ; Chang LIU ; Xiongwei LIU ; Xiu DONG ; Yun CHEN ; Tingting FENG
China Pharmacy 2024;35(3):316-321
OBJECTIVE To analyze the chemical constituents and components absorbed into plasma of the extract of Ardisia crenata and to elucidate its possible pharmacodynamic material basis. METHODS Overall, 12 rats were randomly assigned to the blank group (n=6) and A. crenata group (n=6) by the paired comparison method. The drug was administered once daily in the morning and afternoon for three days. Serum samples were prepared from serum after redosing on 4th day. The UPLC-QE-HF-MS/ MS was used to analyze and identify the chemical constituents in A. crenata extract and serum samples. Compound Discoverer 3.0 was employed for retention time correction, peak identification, and peak extraction. According to the secondary mass spectrometry information, the Thermo mzCloud online and Thermo mzVault local databases, referring to the relevant literature and control quality spectrum information were used to preliminarily identify the chemical constituents and components absorbed into plasma of A. crenata. RESULTS A total of 34 compounds were identified from the extract of A. crenata, mainly coumarins, flavonoids, organic acids, amino acids, including bergenin, quercetin, gallic acid, L-pyroglutamic acid, etc. Besides, 5 components absorbed into plasma were identified from serum samples: L-pyroglutamic acid, syringic acid, bergenin, cinnabar root saponin A, and mycophenolic acid. CONCLUSIONS L-pyroglutamic acid, syringic acid, bergenin, cinnabar root saponin A, and mycophenolic acid may act as the pharmacodynamic material basis of A. crenata.
2.Evaluation of chemiluminescence immunoassay kit for detection of hepatitis D virus IgG antibody
Rongchen YUAN ; Fangming CHENG ; Kuanhui XIANG ; Yongcong LI ; Tianxun HUANG ; Zhenchao TIAN ; Xiongwei LIU ; Xiaozhong WANG ; Zhuanguo WANG ; Yahong MA ; Jing ZHOU ; Erhei DAI ; Chungen QIAN ; Tong LI ; Tao SHEN ; Bangning CHENG
Chinese Journal of Laboratory Medicine 2024;47(3):234-238
Objective:This study evaluates the performance of chemiluminescence assay, which is designed to detect Hepatitis D Virus (HDV) Immunoglobulin G (IgG) antibodies.Methods:A comparative analysis was conducted among chemiluminescence anti-HDV IgG reagent, the magnetic particle-based domestic reagent A and domestic reagent B, and the Robo Gene HDV RNA kit, using 1909 HBsAg-positive plasma samples. This comparison aimed to delineate clinical specificity and detection accuracy. The anti-HDV IgG reagent precision was assessed at three different concentration levels following the Clinical Laboratory Standards Institute EP5-A2 guidelines. The specificity of the assay was validated using 200 HAV IgM positive, 545 HBsAg-positive but anti-HDV IgG-negative, 350 anti HCV positive plasma samples and 200 healthy human blood samples. Additionally, a concordance study was conducted with 545 HBsAg-positive and 37 anti-HDV IgG-positive plasma samples, comparing the anti-HDV IgG reagent against reagent A.Results:1 909 HBsAg-positive plasma samples were tested using 3 anti HDV IgG reagent and 1 HDV RNA reagent, 19 samples were identified as anti-HDV IgG-positive. The anti-HDV IgG demonstrated superior accuracy and specificity. The assay exhibited excellent precision, with intra-assay coefficient of variation (CV) values ranging from 1.57% to 4.30%, and inter-assay CV values between 1.71% and 4.67% for detecting samples at high, medium, and low concentration levels. Concordance with Reagent A showed consistent results in both positive and negative detections.Conclusion:In this study, the anti-HDV IgG reagent (chemiluminescence method) displayed outstanding specificity in detecting clinical samples and exhibited a high conformity rate with commercialized reagents, making it potentially suitable for screening anti-HDV IgG in HBsAg-positive samples.
3.Assessment and preliminary clinical application of a domestic nucleic acid detection reagent for hepatitis D virus
Yongcong LI ; Rongchen YUAN ; Kuanhui XIANG ; Guomin OU ; Tianxun HUANG ; Fangming CHENG ; Zhenchao TIAN ; Xiongwei LIU ; Xiaozhong WANG ; Feng GUO ; Yahong MA ; Jing ZHOU ; Erhei DAI ; Bangning CHENG ; Tong LI ; Tao SHEN ; Chungen QIAN
Chinese Journal of Laboratory Medicine 2024;47(3):239-244
Objective:This study aims to evaluate the quality and explore the preliminary clinical applications of a domestically developed hepatitis D virus nucleic acid quantification reagent (abbreviated as"domestic HDV RNA reagent").Methods:The sensitivity and accuracy of the reagent were evaluated in accordance with the WHO HDV RNA international standard, employing the Bio-Rad CFX Opus 96 real-time fluorescence quantitative PCR analysis system. Serial dilutions of pseudo-viruses or cell culture-derived virus were used to determine the linear range of the domestic HDV RNA reagent. Specificity was assessed using positive samples of HAV, HBV, HCV infection, and HEV national reference materials. Precision was evaluated with samples at both high and low concentrations. In a comparative analysis, 30 HDV IgG positive samples were tested using both the domestic HDV RNA reagent and the RoboGene HDV RNA kit based on the ABI 7500 FAST DX system. The Pearson correlation coefficient (r) was used to examine the correlation between the two reagents.Results:The domestic HDV RNA reagent demonstrated a high sensitivity of up to 6 IU/ml, consistent with that of the comparator reagent. The calibration curve for WHO HDV RNA standards had a slope of -3.286, with an amplification efficiency of 101.6%. The linear detection range spanned from 10 to 10 8 IU/ml for eight HDV genotypes. The domestic HDV RNA reagent exhibited exceptional specificity, without cross-reactivity observed with HAV, HBV, HCV, or HEV. Accuracy assessments at five concentration levels met the required standards, with intra-assay precision coefficient of variation ( CV) ranging from 1.20% to 4.20%, and inter-assay precision CV from 1.20% to 7.90%. The detection results for HDV IgG positive samples were highly correlated with the comparator reagent ( r=0.984, P<0.001), achieving a diagnostic accuracy of 100% compared to sequencing results. Conclusion:In this study, the domestic HDV RNA reagent possesses excellent specificity, accuracy, precision, and a broad linear range, attaining a sensitivity level on par with international reagents of the same type.
4.Hemodialysis bilayer bionic blood vessels developed by the mechanical stimulation of hepatitis B viral X(HBX)gene-transfected hepatic stellate cells
LIU HONGYI ; ZHOU YUANYUAN ; GUO PENG ; ZHENG XIONGWEI ; CHEN WEIBIN ; ZHANG SHICHAO ; FU YU ; ZHOU XU ; WAN ZHENG ; ZHAO BIN ; ZHAO YILIN
Journal of Zhejiang University. Science. B 2024;25(6):499-512
Artificial vascular graft(AVG)fistula is widely used for hemodialysis treatment in patients with renal failure.However,it has poor elasticity and compliance,leading to stenosis and thrombosis.The ideal artificial blood vessel for dialysis should replicate the structure and components of a real artery,which is primarily maintained by collagen in the extracellular matrix(ECM)of arterial cells.Studies have revealed that in hepatitis B virus(HBV)-induced liver fibrosis,hepatic stellate cells(HSCs)become hyperactive and produce excessive ECM fibers.Furthermore,mechanical stimulation can encourage ECM secretion and remodeling of a fiber structure.Based on the above factors,we transfected HSCs with the hepatitis B viral X(HBX)gene for simulating the process of HBV infection.Subsequently,these HBX-HSCs were implanted into a polycaprolactone-polyurethane(PCL-PU)bilayer scaffold in which the inner layer is dense and the outer layer consists of pores,which was mechanically stimulated to promote the secretion of collagen nanofiber from the HBX-HSCs and to facilitate crosslinking with the scaffold.We obtained an ECM-PCL-PU composite bionic blood vessel that could act as access for dialysis after decellularization.Then,the vessel scaffold was implanted into a rabbit's neck arteriovenous fistula model.It exhibited strong tensile strength and smooth blood flow and formed autologous blood vessels in the rabbit's body.Our study demonstrates the use of human cells to create biomimetic dialysis blood vessels,providing a novel approach for creating clinical vascular access for dialysis.
5.Correlation between subclavian artery stenosis disease classification and posterior circulation ischemia
Yang LIU ; Aihua HUANG ; Xiongwei ZHANG ; Ying WANG ; Yonghan LIANG ; Guangyu WANG ; Chenxi TAN ; Feng QIU
Chinese Journal of Neuromedicine 2024;23(11):1094-1099
Objective:To explore the correlation between subclavian artery stenosis disease (SASD) classification and posterior circulation ischemia.Methods:A retrospective study was performed; the clinical data, and Doppler vascular ultrasound and vascular imaging results of 81 SASD patients, admitted to Cerebrovascular Stenosis Diagnosis and Treatment Center, Second Affiliated Hospital of Qiqihar Medical College and Department of Neurology, Rocket Force Specialty Medical Center from May 2018 to August 2023, were collected. SASD was categorized into 2 types (single type and concurrent type) based on the presence or absence of other posterior circulation artery (basilar artery, vertebral artery, or subclavian artery distal segment) stenosis/occlusion, and into 3 groups (non-posterior circulation ischemia group, posterior circulation transient ischemic attack group and posterior circulation cerebral infarction group) based on the presence or absence of posterior circulation ischemia. Blood stealing pathways in different SASD classifications were analyzed, and correlation of SASD classification with posterior circulation ischemia was discussed.Results:Single-type SASD was noted in 44 patients (54.3%), mainly initiating blood stealing through the vertebral artery to the vertebral artery and then to the subclavian artery ( n=26); concurrent-type SASD was noted in 37 patients (45.7%), mainly initiating blood stealing through the occipital artery to the costocervical trunk and then to the subclavian artery ( n=10). Sixty-five patients (80.2%) were into the non-posterior circulation ischemia group, 4 (4.9%) into the posterior circulation transient ischemic attack group and 12 (14.8%) into the posterior circulation cerebral infarction group. Among the 44 patients with single-type SASD, 39 did not have posterior circulation ischemia, and 3 had posterior circulation cerebral infarction. Among the 37 patients with concurrent-type SASD, 26 did not have posterior circulation ischemia, and 9 had posterior circulation cerebral infarction. Conclusion:Initiation of blood stealing in SASD patients is related to SASD classification, and concurrent-type SASD patients trend to have posterior circulation ischemia.
6.Application value of peri-gastric devascularization without dissociation of esophagus for portal hypertension
Mingguo TIAN ; Baoding LI ; Feng LIU ; Xiongwei FAN ; Liyun WANG ; Yang BO ; Yong YANG ; Guangming WU
Chinese Journal of Digestive Surgery 2023;22(12):1484-1489
Objective:To investigate the application value of peri-gastric devasculariza-tion without dissociation of esophagus for portal hypertension.Methods:The retrospective and descriptive study was conducted. The clinical data of 94 patients with portal hypertension who were admitted to three medical centers, including 75 cases in the People′s Hospital of Ningxia Hui Autonomous Region, 12 cases in the People′s Hospital of Wuhai and 7 cases in the People′s Hospital of Wuzhong, from July 2018 to December 2022 were collected. There were 68 males and 26 females, aged 46(range, 21-70)years. All 94 patients underwent peri-gastric devascularization without dissociation of esophagus. Observation indicators: (1) intraoperative condition; (2) postoperative complications; (3) follow-up. Measurement data with normal distribution were represented as Mean± SD, and measure-ment data with skewed distribution were represented as M(range). Count data were described as absolute numbers, and comparison between groups was conducted using the chi-square test or Fisher exact probability. Results:(1) Intraoperative condition. All 94 patients underwent surgery success-fully without operation death, including 82 cases receiving open surgery and 12 cases receiving laparoscopic surgery. The operation time and volume of intraoperative blood loss were (183±85)minutes and 289(range, 158-560)mL, respectively, for the 94 patients. (2) Postoperative complications. Of 94 patients, early portal vein thrombosis occurred in 24 cases, intra-abdominal infection occurred in 2 cases, hepatic encephalopathy occurred in 1 case, pulmonary embolism occurred in 1 case, intra-abdominal hemorrhage requiring operation to stop bleeding occurred in 1 case and pleural effusion requiring drainage occurred in 1 case. All patients with postoperative complications were cured after treatment. None of the 94 patient had postoperative esophageal complications such as odynophagia or dysphagia. (3) Follow-up. All 94 patients were followed up for 38(range, 6-60)months. Of the 45 patients with paraesophageal vein, there were 36 cases of thinner and 9 cases of occlusion of the distal subphrenic paraesophageal vein after surgery, respectively. Cases with esophageal varices disappearance, cases with mild and moderate residual of esophageal varices, cases with severe residual of esophageal varices, cases with recurrence of esophageal varices, cases with esophageal varices bleeding were 7, 70, 9, 4, 4 in the 94 patients after surgery. Cases with esophageal varices disappearance was 7 in the 45 patients with paraesophageal vein, versus 0 in the 49 patients without paraesophageal vein, showing a significant difference between them ( P<0.05). Of 94 patients, 17 cases developed postoperative late portal vein thrombosis and cavernous transformation, 7 cases developed liver cancer, 1 case had hepatic encephalopathy, and 6 cases died. Conclusion:Peri-gastric devascularization without dissociation of esophagus is safe and feasible for the treatment of portal hypertension.
7.Comparison of the efficacy of middle meningeal artery embolization and conventional therapy for chronic subdural hematomas: a propensity score matching study
Zhensheng LIU ; Demao CAO ; Yong SUN ; Tingfeng WEI ; Xiongwei KUANG ; Longjiang ZHOU ; Chenyi WU ; Hongsheng WANG
Chinese Journal of Radiology 2022;56(6):661-666
Objective:To compare the effect of middle meningeal artery embolization (MMAE) versus conventional therapy for chronic subdural hematoma (CSDH).Methods:Retrospective analysis of 38 patients with 48 CSDHs treated with MMAE from May 2019 to May 2021 was performed. Comparisons were made with a conventional treatment for 126 patients with 126 CSDHs from January 2016 to May 2021. The MMAE and conventional treatment patients were matched by the propensity score matching method, and a total of 25 pairs of patients (31 pairs of CSDHs) were successfully matched. The CSDH recurrence, rescue treatment, radiographic follow-up outcome, clinical improvement and complication between the two groups were compared by t test, χ 2 test or Fisher exact probability methods. Results:The rescue treatment rate in MMAE group was significantly lower than that in conventional treatment group [0 (0/31) vs 19.4% (6/31), P=0.024] and the complete resolution rate at 6 months follow-up in MMAE group was significantly higher than that in conventional treatment group [96.8 (30/31) vs 74.2% (23/31), P=0.026]. In terms of CSDH recurrence, there was a trend of lower recurrence in the MMAE group [3.2%(1/31) vs 22.6% (7/31), P=0.053]. The complete resolution rate at 3 months follow-up was 61.3% (19/31) in MMAE group and 45.2% (14/31) in conventional treatment, clinical improvement rate was 92.0% (23/25) in MMAE group and 88.0% (22/23) in conventional treatment, good outcome rate (mRS≤2) was 92.0% (23/25) in MMAE group and 84.0% (21/25) in conventional treatment, complication rate was 0(0/25) in MMAE group and 4.0% (1/25) in conventional treatment, and there were no significant differences in all above-mentioned parameters ( P>0.05). Conclusions:The MMAE may be considered as a safe and effective treatment for CSDH, and MMAE for CSDH is associated with lower trend of recurrence, lower rescue treatment rate and better radiographic follow-up outcome than conventional therapy.
8.Application of a computer-assisted surgery system in precision surgery of mediastinal tumors in children
Yao LIU ; Xiongwei WU ; Yuhe DUAN ; Chunyang ZHAO ; Feifei WANG ; Fengjiao WANG ; Nan XIA ; Xiwei HAO ; Qian DONG
Chinese Journal of Applied Clinical Pediatrics 2022;37(22):1750-1755
Objective:To analyze and summarize the application and significance of the computer-assisted surgery system in precision surgery of mediastinal tumors in children.Methods:The clinical data of 54 children that underwent mediastinal tumor resection surgery in the Affiliated Hospital of Qingdao University from August 2008 to November 2021 were collected.According to whether the Hisense CAS computer-assisted surgery system was used before the operation, the patients were divided into three-dimensional (3D) reconstruction group (29 cases, 53.70%) and two-dimensional (2D) CT group (25 cases, 46.30%). The surgical indicators, postoperative hospital stay, and the incidence of surgical complications were compared between the two groups.The t-test, Mann- Whitney U test, χ2 test or Fisher′ s exact test were carried out for statistical analysis.Simulated surgery and actual surgery were compared for both 3D reconstruction and 2D CT groups. Results:There were no significant differences in gender, age, height, weight, maximum tumor diameter and tumor location between the 3D reconstruction group and 2D CT group (all P>0.05). The operation time of the 3D reconstruction group was [(125.14±41.37) min] was shorter than that of the 2D CT group [(149.24±44.53) min] ( P=0.044). The intraoperative blood loss in the 3D reconstruction group [15.00(13.50, 25.00) mL] was less than that in the 2D CT group [36.00(30.00, 75.00) mL] ( P<0.001). In addition, the indwelling time the closed thoracic drainage tube [4.00 (3.00, 5.50) d] and postoperative hospitalization days [(8.83±3.39) d] in the 3D reconstruction group were shorter than those in the 2D CT group [7.00(5.00, 11.50) d, (11.00±4.10) d] ( P=0.001, 0.038). No significant difference in postoperative complication rates was found between the 3D reconstruction group and 2D CT group ( P>0.05). The simulated operation was consistent with the actual operation in the 3D reconstruction group. Conclusions:3D reconstruction by the computer-assisted surgery system can truly reveal the anatomical relationship between tumors and surrounding organs and blood vessels, and improve the accuracy and safety of surgical resection of mediastinal tumors in children.
9.Analysis of Chemical Constituents as Flavonoids and Coumarins in Radix Ardisiae from Different Sources
Xiao LI ; Hui SHI ; Jingxin DING ; Tingting FENG ; Xiongwei LIU ; Chang LIU ; Ying ZHOU
China Pharmacy 2021;32(4):443-452
OBJECTIVE:To identif y and analyze the flavonoids and coumarins in Radix Ardisiae from different sources. METHODS:UPLC-QE-HF-MS/MS was adopted. The determination was performed on Zorbax Eclipse-C 18 column with mobile phase consisted of acetonitrile- 0.1% formic acid solution (gradient elution )at the flow rate of 0.3 mL/min. The column temperature was 30 ℃,and the temperature of injector was 4 ℃. The sample size was 2 µL;ESI source was applied in negative and positive scanning ion mode ,the heater temperature was 325 ℃,the sheath gas pressure was 45 arb,the auxiliary gas pressure was 15 arb,the purge gas pressure was 1 arb,the electrospray voltage was 3.5 kV,the capillary temperature was 330 ℃, S-lens RF level was 55%,scan mode was first-order full sca m/z 100-1 500,data-dependent secondary mass spectrometry scanning (dd-MS2,Top N =10),the resolution was 70 000 (first mass spectrometry ) , 17 500 (secondary mass spectrometry),the collision mode was high-energy collision dissociation. Through retrieving foreign and domestic databases as ChemSpider ,mzCloud,mzVault,PubChem,the structure of the compound was identified on the basis of related literatures and reference data ,and the conten ts were compared. RESULTS & CONCLUSIONS:A total of 47 components were separated from Radix Ardisiae of 3 kinds of sources as Ardisia crenata Sims,A. crispa(Thunb.)A. DC. ,A. crenata Sims var . bicolor (Walk)C. Y. Wu et C. Chen. A total of 17 flavonoids were identified ,including 9 flavonols (quercetin 3-O-rhamnoside-7-O-glucoside, myricetin, rutin, mauritanin, kaempferol, quercetin, isorhamnetin, quercetin,mearnsitrin),3 flavan-3-ols [(-)-epigallocatechin,catechin,epigallocatechin gallate )2 dihydroflavonoids [fustin , eriodictyol] and 3 other types [ 3-(2,3-dihydro-benzo[1,4]dioxin-6-yl)-7-hydroxy-2-trifluoromethyl-chromen-4-one,methadone, oriciacridone F] ,10 coumarins {bergenin ,([ 7-hydroxy-4-methyl-2-oxo-2H-chromen-6-yl)oxy]acetic acid ,[7-(carboxymethoxy)- 4-methyl-2-oxo-2hydroxychromo-3-yl]acetic acid ,4,9-dihydroxy-7H-furo[3,2-g]chromen-7-one,6,7-dihydroxy-4-methylcoumarin, esculetin,fraxetin,7,8-dihydroxy-4-methylcoumarin,4-methylumbelliferyl glucuronide ,scoparone}. Results of content analysis showed that in flavonoids and coumarins ,there were 5 common components in Radix Ardisiae from 3 kinds of sources ,i.e. bergenin(peak 2),[7-(carboxymethoxy)-4-methyl-2-oxo-2-hydroxychromo-3-yl] acetic acid (peak 5),methadone(peak 16), quercetin(peak 18),oriciacridone F (peak 26);the contents of common components were significantly different. In addition to 5 common components ,there were 22 different chemical components ,which were compounds corresponding to peaks 1,3,4, 6-15,17,19-25 and 27,respectively. Among them ,compounds corresponding to peaks 3,6,8 and 23 were only found in A. crenata Sims var. bicolor(Walk)C. Y. Wu et C. Chen ;compounds corresponding to peaks 12-15,19 were only found in A. crispa (Thunb.)A. DC. UPLC-QE-HF-MS/MS method can efficiently ,accurately and quickly identify the flavonoids and coumarins in Radix Ardisiae from different sources.
10.Comparative study on preoperative planning of pediatric hepatoblastoma between three-dimensional and two-dimensional imaging of computer-assisted surgery system
Xiongwei WU ; Jie LIU ; Linlin WU ; Xin CHEN ; Fujiang LI ; Yuhe DUAN ; Qian DONG
Chinese Journal of General Surgery 2020;35(9):673-676
Objective:To compare the application of preoperative planning of pediatric hepatoblastoma between three-dimensional and two-dimensional imaging of computer-assisted surgery system.Methods:From Jan 2016 to Oct 2019, surgical resection plans on 25 cases were made by the operator team through the observation of two-dimensional CT images. Then, the team will make a thorough analysis and revise the previous preoperative plan for the children patient according to three-dimensional imaging of the computer-assisted surgery system.Results:All the patients had the tumor successfully removed and achieved R 0 resection based on the preoperative plan assisted by three-dimensional imaging. All the children patients were doing well as found by postoperative short-term follow-up. Among them, preoperative plans were finally revised in 10 cases due to the disparity with the original ones in three-dimensional relationship between tumor and blood vessels, in vascular variation and in the volume of remnant liver after the evaluation of three-dimensional reconstruction images, which account for 40% variation of the original operation scheme. Conclusions:Three dimensional imaging of computer-assisted surgery system is superior to two-dimensional imaging in preoperative planning of pediatric hepatoblastoma.

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