1.Qualitative and Quantitative Analysis of Chemical Constituents in Gualou Niubangtang by UPLC-Q-TOF-MS/MS and HPLC
Yiyi ZHANG ; Jing YANG ; Yuqing CHENG ; Huimin GAO ; Jin QIN ; Li YAO ; Xiyang DU ; Raorao LI
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(6):179-187
ObjectiveThis paper aims to clarify the material basis of Gualou Niubangtang and establish a quantitative analysis method for its main constituents, providing a reference for the overall quality control of this preparation. MethodsThe constituents in the formula were systematically characterized based on ultra-performance liquid chromatography-quadrupole time-of-flight tandem mass spectrometry (UPLC-Q-TOF-MS/MS). Identification was performed by matching with the UNIFI 9.6 software and utilizing database platforms such as PubChem, ChemicalBook, and ChemSpider, combined with relevant literature reports. A quantitative analysis method for the seven main constituents in Gualou Niubangtang was established by using high performance liquid chromatography (HPLC). ResultsUPLC-Q-TOF-MS/MS analysis identified 155 constituents, including 69 flavonoids, 36 terpenoids, 23 phenylpropanoids, 8 phenylethanoid glycosides, and 19 other types of constituents. In the established quantitative analysis method, the seven main constituents showed good linearity within their respective linear ranges. The precision, repeatability, stability, and spike recovery all met the required standards. The results showed that the content ranges of geniposide, liquiritin, hesperidin, arctiin, baicalin, oroxylin A-7-O-β-D-glucuronide, and wogonoside in 15 batches of Gualou Niubangtang were 13.67-21.25, 1.20-7.64, 5.45-7.45, 22.97-33.51, 29.95-39.07, 2.58-4.80, and 6.56-9.31 mg·g-1, respectively. ConclusionThis study successfully characterizes and attributes multi-category constituents in Gualou Niubangtang, clarifying that its material basis is primarily composed of flavonoids, terpenoids, phenylethanoid glycosides, and phenylpropanoids. Furthermore, it enables the quantification of seven constituents within the formula. This work lays a foundation for research on the quality control, action mechanism, and clinical application of this formula.
2.Clinical application of KASP-based RHCE genotyping in RhD-positive patients
Xiaoyu LIAN ; Mengdan LI ; Xiaoyu GUAN ; Li TIAN ; Chenying WANG ; Di WU ; Tianqiong LUO ; Xiaolin DU ; Xin JI ; Haixia XU ; Jue WANG ; Ling LI ; Zhong LIU
Chinese Journal of Blood Transfusion 2026;39(5):596-602
Objective: To develop a RHCE genotyping assay based on kompetitive allele-specific PCR (KASP) and assess its clinical accuracy for RhCE blood group determination. Methods: KASP primers were designed to interrogate three RHCE loci: the 109 bp insertion/deletion in intron 2, c. 307T>C, and c. 676C>G. A total of 1 194 RhD-positive inpatients from Chengdu were typed by both KASP genotyping and manual tube serology. Discordant samples (n=10) were retested by both methods and further resolved by Sanger sequencing. An additional 377 cases were tested for the c. 48C>G locus to evaluate the predictive accuracy of individual loci and combined locus testing for RhC antigen. Results: Genotyping concordance with serology was 100.0% for both the c. 676C>G locus (RhE/Rhe) and the c. 307T>C locus (Rhc). For RhC prediction using the 109 bp insertion, overall accuracy was 99.7% (1 191/1 194); the 3 discordant cases were confirmed by Sanger sequencing to be false negatives attributable to 109 bp deletion in intron 2. Testing the c. 48C>G allele for RhC prediction yielded 7 false positives, with an accuracy of 98.1% (370/377). RhC antigen status was determined by combining the 109 bp insertion and the c. 48C allele. After excluding 10 samples with inconsistent results between the two loci, the accuracy reached 100% in the remaining 367 samples. When both loci were applied in combination, accuracy reached 100% in the 367 cases with concordant results. Among the 1 194 patients, CCee (45.8%) and CcEe (31.7%) were the most common RhCE phenotypes. The e antigen had the highest positivity rate (92.2%), and the Ce haplotype was the most frequent (66.9%). Conclusion: The KASP-based RHCE genotyping method achieves high accuracy for clinical RhCE typing. Combining the 109 bp insertion/deletion with the c. 48C allele significantly improves RhC antigen prediction compared with either locus alone. This method was applied to RhCE genotyping of 1 194 RhD-positive inpatients in Chengdu, providing local RhCE phenotype and haplotype distribution data to support RhCE-matched transfusion practice.
3.Multimodal Assessment of a Hypoxic Pulmonary Hypertension Mouse Model
Hongzhi DU ; Jin FAN ; Xiaolu WEI ; Lianmei WANG ; Yan LIU ; Weiya CHEN ; Tengfei CHEN ; Yunhang GAO ; Ling SONG ; Guangping ZHANG ; Hongping HOU
Laboratory Animal and Comparative Medicine 2026;46(4):476-486
ObjectiveTo establish a mouse model of hypoxic pulmonary hypertension (HPH) through intermittent hypoxia induction, and to develop a comprehensive and reliable evaluation system for the HPH model, thereby providing experimental evidence for mechanistic studies and translational applications related to this disease. MethodsTwenty-four male specific pathogen-free (SPF) BALB/c mice were randomly divided into a control group and a model group, with 12 mice per group. Mice in the model group were placed in a hypobaric oxygen chamber control system to establish the HPH model, whereas mice in the control group received no intervention. After 28 days of modeling, a comprehensive evaluation of the pathophysiological characteristics of both groups was performed using a general condition scoring scale, echocardiography, hemodynamic measurements, blood gas analysis, hematological tests, organ coefficient determination, histopathological examination, and enzyme-linked immunosorbent assay (ELISA). In addition, correlation analyses were conducted among echocardiographic parameters, the contents of endothelin‑1 (ET‑1) and N‑terminal pro‑B‑type natriuretic peptide (NT‑proBNP) in lung tissue, and other measured indices. ResultsWith prolonged modeling duration time, body weight, water intake, and activity level of mice in the model group were significantly decreased compared with the control group (P<0.05). Echocardiography showed that, compared with the control group, the model group exhibited increased right ventricular dimensions (P<0.01), right ventricular anterior wall thickening (P<0.01), and widened main pulmonary artery diameter (P<0.01), whereas the peak systolic velocity across the pulmonary valve, tricuspid annular plane systolic excursion, and peak systolic velocity of the tricuspid annulus were significantly decreased (P<0.001); tricuspid regurgitation was observed in some model animals. Hemodynamic results revealed that right ventricular systolic pressure was elevated in the model group compared with the control group (P<0.001). Blood gas analysis showed that pH, partial pressure of oxygen, oxygen saturation, actual bicarbonate, and total carbon dioxide were all decreased in the model group compared with the control group (P<0.01). Hematological indices demonstrated that lymphocyte counts (P<0.05) and reticulocyte counts (P<0.001) were decreased in the model group compared with the control group. Compared with the control group, the organ coefficients of heart and lung in the model group were significantly increased (P<0.05 and P<0.001). Pathological examination revealed that the right ventricular hypertrophy index was significantly increased in the model group (P<0.001), with varying degrees of damage to the right ventricle, pulmonary artery, and pulmonary vessels; both the pulmonary artery wall thickness percentage and the pulmonary wall area percentage were significantly elevated (P<0.001). ELISA results showed that the levels of ET‑1 and NT‑proBNP in lung tissues were significantly increased in the model group compared with the control group (P<0.05 and P<0.001, respectively). Correlation analysis indicated that some echocardiographic parameters were highly correlated with multiple indices in the development of HPH (P<0.05). ConclusionEchocardiography can accurately assess a series of hemodynamic changes in HPH, including right ventricular structural, functional impairment, and pulmonary hypertension. Laboratory tests not only help verify whether the model has been successfully established, but also provide deeper insights into the pathogenesis of HPH, evaluate the effects of interventions, and offer scientific evidence for clinical outcomes. Pathological examination can further confirm the alterations in pulmonary vascular remodeling and increased right heart load. This multimodal analysis provides a reliable animal model and evaluation paradigm for both basic and translational research on HPH, and is of great significance for exploring disease mechanisms and developing precision therapeutic strategies.
4.Mechanism of Huangqi Xiayuxue Decoction Against Hepatocellular Carcinoma via PI3K/Akt/FoxO Signaling Pathway
Xingru XING ; Yuanzhou SHI ; Jiawei WANG ; Di WU ; Yuyao ZHANG ; Yalin WU ; Du CHEN ; Zhen ZHANG ; Sha TIAN
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(20):131-141
ObjectiveTo explore the therapeutic effect and mechanism of Huangqi Xiayuxue decoction against hepatocellular carcinoma based on network pharmacology, animal experiments, and cell experiments. MethodsThe active ingredients and corresponding targets of Huangqi Xiayuxue decoction were screened via databases including the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) and Bioinformatics Analysis Tool for Molecular Mechanism of Traditional Chinese Medicine (BATMAN-TCM). Hepatocellular carcinoma-related targets were retrieved from the GeneCards database. The common targets shared by Huangqi Xiayuxue decoction and hepatocellular carcinoma were subjected to Gene Ontology (GO) functional annotation and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses. Huangqi Xiayuxue decoction-containing plasma was prepared. The cell counting kit-8 (CCK-8) assay was adopted to observe the effects of different volume fractions (5%, 10%, 15%, and 20%) of Huangqi Xiayuxue decoction-containing plasma on the proliferation of HepG2, MHCC-97H, Hepa1-6, and Huh-7 cells, and the concentration for subsequent experiments was determined. Flow cytometry was adopted to detect the effects of Huangqi Xiayuxue decoction-containing plasma (5%, 10%, 15%, and 20%) on the apoptosis of MHCC-97H cells. A mouse model of subcutaneous xenograft hepatocellular carcinoma was established. The model mice were assigned into low-, medium-, and high-dose (4.63, 9.25, 18.5 g·kg-1) Huangqi Xiayuxue decoction and sorafenib (20 mg·kg-1) groups. Body mass and tumor volume of mice were dynamically monitored during the intervention period. After 3 weeks of intervention, hematoxylin-eosin (HE) staining was performed to observe tumor morphology. Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) assay was employed to detect cell apoptosis. Immunohistochemistry (IHC) was adopted to examine the expression of cysteinyl aspartate-specific proteinase-3 (Caspase-3), B-cell lymphoma-2 (Bcl-2), and Bcl-2-associated X protein (Bax) in the tumor tissue. Real-time PCR and Western blot were employed to verify the changes in apoptosis-related factors and the phosphatidylinositol 3-kinase/protein kinase B/forkhead box O1 (PI3K/Akt/FoxO1) signaling pathway. ResultsA total of 220 common targets shared by Huangqi Xiayuxue decoction and hepatocellular carcinoma were screened out by network pharmacology, among which tumor protein p53 (TP53), Akt1, and signal transducer and activator of transcription 3 (STAT3) were the core targets. Pathway enrichment analysis indicated that the common targets were involved in cell apoptosis and proliferation, as well as the PI3K/Akt and FoxO signaling pathways. Cell experiments showed that compared with the blank plasma group, all volume fractions of Huangqi Xiayuxue decoction-containing plasma inhibited the proliferation of HepG2, MHCC-97H, Hepa1-6, and Huh-7 cells (P<0.01). The proportion of early apoptotic cells in MHCC-97H cells was increased after treatment with different volume fractions of Huangqi Xiayuxue decoction-containing plasma (P<0.01). Animal experiments revealed that Huangqi Xiayuxue decoction inhibited the growth of tumor volume, promoted tumor cell apoptosis in the high-dose group, up-regulated the protein and mRNA levels of Bax and Caspase-3, down-regulated the protein and mRNA levels of Bcl-2, and reduced the phosphorylation level of the PI3K/Akt/FoxO1 signaling pathway (P<0.05, P<0.01). ConclusionHuangqi Xiayuxue decoction exerts anti-hepatocellular carcinoma effects by inhibiting the activation of the PI3K/Akt/FoxO1 signaling pathway and promoting hepatocellular carcinoma cell apoptosis.
5.Mechanism of Huangqi Xiayuxue Decoction Against Hepatocellular Carcinoma via PI3K/Akt/FoxO Signaling Pathway
Xingru XING ; Yuanzhou SHI ; Jiawei WANG ; Di WU ; Yuyao ZHANG ; Yalin WU ; Du CHEN ; Zhen ZHANG ; Sha TIAN
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(20):131-141
ObjectiveTo explore the therapeutic effect and mechanism of Huangqi Xiayuxue decoction against hepatocellular carcinoma based on network pharmacology, animal experiments, and cell experiments. MethodsThe active ingredients and corresponding targets of Huangqi Xiayuxue decoction were screened via databases including the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) and Bioinformatics Analysis Tool for Molecular Mechanism of Traditional Chinese Medicine (BATMAN-TCM). Hepatocellular carcinoma-related targets were retrieved from the GeneCards database. The common targets shared by Huangqi Xiayuxue decoction and hepatocellular carcinoma were subjected to Gene Ontology (GO) functional annotation and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses. Huangqi Xiayuxue decoction-containing plasma was prepared. The cell counting kit-8 (CCK-8) assay was adopted to observe the effects of different volume fractions (5%, 10%, 15%, and 20%) of Huangqi Xiayuxue decoction-containing plasma on the proliferation of HepG2, MHCC-97H, Hepa1-6, and Huh-7 cells, and the concentration for subsequent experiments was determined. Flow cytometry was adopted to detect the effects of Huangqi Xiayuxue decoction-containing plasma (5%, 10%, 15%, and 20%) on the apoptosis of MHCC-97H cells. A mouse model of subcutaneous xenograft hepatocellular carcinoma was established. The model mice were assigned into low-, medium-, and high-dose (4.63, 9.25, 18.5 g·kg-1) Huangqi Xiayuxue decoction and sorafenib (20 mg·kg-1) groups. Body mass and tumor volume of mice were dynamically monitored during the intervention period. After 3 weeks of intervention, hematoxylin-eosin (HE) staining was performed to observe tumor morphology. Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) assay was employed to detect cell apoptosis. Immunohistochemistry (IHC) was adopted to examine the expression of cysteinyl aspartate-specific proteinase-3 (Caspase-3), B-cell lymphoma-2 (Bcl-2), and Bcl-2-associated X protein (Bax) in the tumor tissue. Real-time PCR and Western blot were employed to verify the changes in apoptosis-related factors and the phosphatidylinositol 3-kinase/protein kinase B/forkhead box O1 (PI3K/Akt/FoxO1) signaling pathway. ResultsA total of 220 common targets shared by Huangqi Xiayuxue decoction and hepatocellular carcinoma were screened out by network pharmacology, among which tumor protein p53 (TP53), Akt1, and signal transducer and activator of transcription 3 (STAT3) were the core targets. Pathway enrichment analysis indicated that the common targets were involved in cell apoptosis and proliferation, as well as the PI3K/Akt and FoxO signaling pathways. Cell experiments showed that compared with the blank plasma group, all volume fractions of Huangqi Xiayuxue decoction-containing plasma inhibited the proliferation of HepG2, MHCC-97H, Hepa1-6, and Huh-7 cells (P<0.01). The proportion of early apoptotic cells in MHCC-97H cells was increased after treatment with different volume fractions of Huangqi Xiayuxue decoction-containing plasma (P<0.01). Animal experiments revealed that Huangqi Xiayuxue decoction inhibited the growth of tumor volume, promoted tumor cell apoptosis in the high-dose group, up-regulated the protein and mRNA levels of Bax and Caspase-3, down-regulated the protein and mRNA levels of Bcl-2, and reduced the phosphorylation level of the PI3K/Akt/FoxO1 signaling pathway (P<0.05, P<0.01). ConclusionHuangqi Xiayuxue decoction exerts anti-hepatocellular carcinoma effects by inhibiting the activation of the PI3K/Akt/FoxO1 signaling pathway and promoting hepatocellular carcinoma cell apoptosis.
6.Risk factors of acute kidney injury induced by vancomycin in pediatric patients: a meta-analysis
Mingfu TUO ; Xiaoyuan DI ; Kun YANG ; Caie TANG ; Yan DU ; Hongying HE
Adverse Drug Reactions Journal 2025;27(6):332-338
Objective:To systematically evaluate the incidence and risk factors of acute kidney injury (AKI) induced by vancomycin in pediatric patients.Methods:Databases of PubMed, Embase, The Cochrane Library, Web of Science, CNKI, Wanfang, VIP, Chinese Biomedical Database (CBM) were searched and articles about the risk factors of AKI induced by vancomycin in pediatric patients from inception to June 2024 were collected. Quality assessment was performed using the Newcastle-Ottawa Scale (NOS) for the included studies. Meta-analysis of the data for relevant exposure factors extracted from the included literature was conducted using Rev Man 5.4. The strength of association between the exposure factors and AKI was expressed using the odds ratio ( OR) and its 95% confidence interval ( CI). Results:A total of 13 studies were entered, involving 11 073 patients. Of them, 1 388 patients were in AKI group and 9 685 patients in non-AKI group. The incidence of AKI was 12.53%, ranging from 4.62% to 27.07%. The quality evaluation results showed that the 13 documents were all of high-quality (NOS score ≥7 points). Meta-analysis showed that admission to intensive care unit (ICU) ( OR=2.39, 95% CI: 1.59-3.59, P<0.001), vancomycin using time ≥7 d ( OR=2.19, 95% CI: 1.44-3.34 P=0.003), vancomycin steady-state trough concentration ≥15 mg/L ( OR=2.98, 95% CI: 2.22-4.01, P<0.001), combined with nephrotoxic drugs ≥2 kinds ( OR=2.92, 95% CI=1.84-4.64, P<0.001), combined with piperacillin sodium and tazobactam sodium ( OR=2.71, 95% CI: 1.72- 4.27, P<0.001), combined with carbapenem ( OR=2.36, 95% CI: 1.36-4.10, P=0.002), combined with aminoglycosides ( OR=1.78, 95% CI: 1.35-2.35, P<0.001), combined with loop diuretics ( OR=3.16, 95% CI: 2.36- 4.23, P<0.001), combined with amphotericin B ( OR=2.26, 95% CI: 1.35-3.79, P=0.002), combined with contrast medium ( OR=2.34, 95% CI: 1.04-5.25, P=0.040), and combined with aciclovir ( OR=1.74, 95% CI: 1.04-2.84, P=0.030) were all risk factors of AKI induced by vancomycin in pediatric patients. Conclusions:The incidence of vancomycin-related AKI in pediatric patients was 12.53%. Admission to ICU, vancomycin trough concentration ≥15 mg/L, medication time ≥7 d, and concomitant use of ≥2 nephrotoxic drugs and etc.were risk factors of vancomycin-related AKI.
7.Clinical study of intracranial hypotension targeted body posture combined with pharmacotherapy in the treatment of chronic subdural hematoma
Jiayu CHEN ; Zhe WANG ; Di ZANG ; Ruizhe ZHENG ; Xiangru YE ; Zengxin QI ; Zeyu XU ; Zhiqiang LI ; Chengfeng SUN ; Liangjun SHEN ; Luoping SHENG ; Fulin XU ; Ruyong YE ; Kaiyu ZHOU ; Weijun TANG ; Yueqing HU ; Dapeng SHI ; Yuquan WANG ; Xizhen WU ; Ying WANG ; Qilin ZHANG ; Feili LIU ; Guo YU ; Yiping LU ; Yirui SUN ; Ning ZHANG ; Feng HUANG ; Xialong GU ; Han ZHANG ; Jian DING ; Yongyan BI ; Haolan DU ; Jing ZHANG ; Hailong JI ; Ding DING ; Wei ZHANG ; Xuehai WU
Chinese Journal of Surgery 2025;63(3):212-218
Objective:To compare the efficacy of body posture combined with pharmacotherapy and pharmacotherapy alone in the treatment of chronic subdural hematoma(CSDH).Methods:Firstly, retrospective case series study was conducted. Thirty cases of CSDH that had received body posture combined with pharmacotherapy at Department of Neurosurgery, Huashan Hospital Affiliated to Fudan University from December 2016 to October 2020 were studied retrospectively. Twenty-seven patients were male, and 3 patients were female. The age of patients ( M(IQR)) was 66(16) years (range:28 to 84). Nineteen patients had unilateral hematoma, and 11 patients had bilateral hematoma. All patients received pharmacotherapy and body posture therapy that was to raise their lower limbs 20 to 30 cm with leg lift pad and get abdominal compressed with customized abdominal belt in supine position. Patients were required to maintain the body posture as much as possible, with the maximum to 16 to 18 hours per day. Patients with unilateral hematoma should tilt the head to the affected side and avoid tilting it to the opposite side. For patients with bilateral hematoma, there was no need for head lateralization. Patient were treated with oral dexamethasone and atorvastatin simultaneously. The preliminary efficacy of body posture combined with pharmacotherapy was determined by hematoma improvement rate which was analyzed by Clopper-Pearson method. Then, the multi-center, prospective, randomized controlled trial had carried out in 9 medical centers from August 2020 to November 2021. The stratified block randomization method was adopted. Patients were randomized in a ratio of 1∶1 to either receive pharmacotherapy alone(the control group) or body posture combined with pharmacotherapy(the experiment group) for 3 months and followed up for 6 months. Effective treatment was defined as complete absorption of hematoma, or the hematoma volume decreased by more than 10 ml and Markwalder grading scale score had improved by more than 1 point compared to the baseline. The efficacy rate and surgery conversion rate at 3 months and recurrence at 6 months were observed. Comparison between groups was performed with paired sample t test, Mann-Whitney U test, χ2 test, corrected χ2 test, or Fisher exact probability method. Logistic regression was used to compare the effective rate and operation rate between the two groups. Results:In the respective study, 30 patients completed follow-up 13 to 353 days after treatment. At the last follow-up, the incidence of almost complete absorption or significantly absorption of hematoma (hematoma volume was significantly reduced accompanied by symptom improvement) was 93.3%. The 95% CI for the incidence that analyzed by the Clopper-Pearson method was 77.9% to 99.2%. One hundred and six patients were enrolled in the multicenter study. Fifty-five patients underwent body posture combined with pharmacotherapy. The age was 74(17) years (range:26 to 92). Thirty-nine patients were males and 16 were females. Fifty-one patients underwent pharmacotherapy alone. The age was 69(12) years (range:48 to 84). Thirty-seven patients were males and 14 were females. The length of body posture recorded in diary card was (15.7±2.3) hours(range:7.6 to 19.3 hours). The efficacy rate in the body posture combined with pharmacotherapy group and pharmacotherapy alone group were 83.6% (46/55) and 56.9% (29/51), respectively at 3 months. The result of the logistic regression analysis showed that the efficacy of body posture combined with pharmacotherapy group was better than that of pharmacotherapy alone group ( OR=3.88,95% CI:1.57 to 9.58, P=0.003). Surgery rate in the body posture combined with pharmacotherapy group and pharmacotherapy alone group were 5.5% (3/55) and 21.6% (11/51) respectively. The result of Logistic regression showed that the pharmacotherapy alone group was more likely to be converted to surgery ( OR=0.21,95% CI:0.05 to 0.80, P=0.023). At the 6 months, no recurrence of cases was found in the body posture combined with pharmacotherapy group. However, the recurrence rate of pharmacotherapy alone group was 6.3% (3/48), there was no significant difference between the two groups ( P>0.05). Conclusion:The effect of body posture combined with pharmacotherapy for chronic subdural hematoma is better than that of pharmacotherapy alone.
8.Establishment of reference values for clot waveform analysis parameters and their clinical application in differentiating acquired hemophilia A from lupus anticoagulant positive conditions
Bin YAN ; Mengchao CUI ; Yuanyuan WEN ; Di WU ; Luyi RU ; Huixin ZOU ; Tianxi HU ; Ruijuan WANG ; Suping ZHAI ; Weipeng DU
Chinese Journal of Laboratory Medicine 2025;48(3):371-377
Objective:To establish reference values for clot waveform analysis (CWA) and analyze their diagnostic efficacy in distinguishing acquired hemophilia A (AHA) and lupus anticoagulant (LA)-positive patients.Methods:Case-Control Study. A total of 391 healthy individuals(260 males and 131 females) with a mean age of 45.53±14.85 years were enrolled at Nanyang central Hospital between January 6, 2023 and October 10, 2024. Prothrombin time (PT), activated partial thromboplastin time (APTT), and thrombin time (TT) were measured to establish reference ranges for the CWA parameters, including maximal reaction velocity (Min1), maximal reaction acceleration (Min2), and maximal reaction deceleration (Max2). A total of 158 definitively diagnosed AHA and LA-positive patients (mean age:42.46±14.83 years), including 34 AHA patients and 124 LA-positive patients, were recruited. The Mann Whitney U test was used to analyze the differences in the CWA parameters between the two groups. The diagnostic efficacy of CWA parameters in distinguishing AHA and LA-positive patients was evaluated using the area under the receiver operating characteristic(ROC) curve AUC and the cut-off values were calculated. Results:The reference values for PT-Min1, APTT-Min1, APTT-Min2, APTT-Max2, TT-Min1, TT-Min2, TT-Max2 were 203.41-516.89, 144.63-324.03, 526.46-1 190.03, -404.96±157.22, 159.17±60.34, 272.29-686.99, and -289.47--113.76, respectively. Compared with the CWA parameters in AHA patients, APTT-Max2 was significantly lower in LA-positive patients [-422.74(-577.50, -239.22) vs. -68.87(-92.85,30.28), Z=-7.43, P<0.01], while PT-Min1, APTT-Min1, APTT-Min2, TT-Min1, TT-Min2 were significantly elevated [287.01(188.03, 382.50) vs. 107.45(90.20, 151.39), 972.88(601.20, 1 351.19) vs. 229.10(118.38, 371.67), Z=6.68, 6.69, all P<0.01]. ROC analysis demonstrated the APTT-CWA parameter exhibited high diagnostic efficacy in patients with AHA (AUC>0.900 for both).Additionally, APTT-Min1 and APTT-Max2 were found to be useful in distinguishing between AHA patients and those with LA-positive status accompanied by APTT prolongation (AUC=0.660, 0.700, respectively). Conclusions:Reference values for CWA parameters were successfully established. The APTT-CWA is useful for differentiating between AHA and LA-positive patients and APTT-Max2 demonstrated a good diagnostic value in differentiating AHA patients from those with LA-positive status accompanied by APTT prolongation.
9.Mechanism of curcumin on improving cell damage induced by ultraviolet B irradiation
Ying CHEN ; Ju-hua ZHAO ; Yu YANG ; Xiu-jun DU ; Hai-xia LIU ; Ling-ling XIONG ; Hua-di ZHUANG
Journal of Regional Anatomy and Operative Surgery 2025;34(9):753-758
Objective To explore the effect of curcumin(CUR)on oxidative damage of keratinocytes induced by ultraviolet B(UVB)irradiation through Toll-like receptor 4(TLR4)/nuclear factor-kappa B(NF-κB)/nucleotide-binding oligomerization domain-containing protein 3(NLRP3)signaling pathway.Methods Human keratinocytes of HaCaT were cultured normally in vitro,and the keratinocyte oxidative damage model was established by the irradiation of 57 mJ/cm2 UVB.The cells with normal culture were as the control group,the cells treated after modeling were as the UVB group,the cells treated with 5 μmol/L CUR after modeling were as the CUR group,the cells treated with 100 μg/L TLR4 inhibitor of TAK-242 after modeling were as the TAK-242 group,and the cells treated with 5 μmol/L CUR and 100 nmol/L TLR4 activator of lipopolysaccharide(LPS)were as the CUR+LPS group.qRT-PCR was applied to detect the relative expression levels of TLR4,NF-κB,and NLRP3 mRNAs of cells in each group.CCK-8 was applied to detect the cell proliferation in each group.The relative content of reactive oxygen species(ROS),the viabilities of superoxide dismutase(SOD)and catalase(CAT),and the concentrations of glutathione(MDA)and glutathione(GSH)of cells in each group were detected by fluorescence assay according to the kit instruction.ELISA kit was used to detect the expression of inflammatory factors of tumor necrosis factor-α(TNF-α)and interleukin-1β(IL-1β)of cells in each group.Flow cytometry was applied to detect the cell apoptosis in each group.Western blot was applied to detect the expression of proliferation related protein of proliferating cell nuclear antigen(PCNA),apoptosis related proteins[B-cell lymphoma-2(Bcl-2)and Bcl-2-associated X protein(Bax)],and TLR4/NF-κB/NLRP3 signaling pathway related proteins(TLR4,NF-κB and NLRP3)of cells in each group.Results Compared with the Control group,the cell survival rate,the expression levels of PCNA and Bcl-2 proteins,the viabilities of SOD and CAT,and the GSH concentration in the UVB group decreased,while the apoptosis rate,the level of Bax protein,the relative content of ROS,the concentration of MDA,the levels of TNF-α and IL-1β,and the mRNA and protein levels of TLR4,NF-κB and NLRP3 increased(P<0.05).Compared with the UVB group,the cell survival rate,the expression levels of PCNA and Bcl-2 proteins,the viabilities of SOD and CAT,and the GSH concentration in the TAK-242 group and CUR group increased,while the apoptosis rate,the level of Bax protein,the relative content of ROS,the concentration of MDA,the levels of TNF-α and IL-1β,and the mRNA and protein levels of TLR4,NF-κB and NLRP3 decreased(P<0.05).Compared with the CUR group,the cell survival rate,the expression levels of PCNA and Bcl-2 proteins,the viabilities of SOD and CAT,and the GSH concentration in the CUR+LPS group decreased,while the apoptosis rate,the level of Bax protein,the relative content of ROS,the concentration of MDA,the levels of TNF-α and IL-1β,and the mRNA and protein levels of TLR4,NF-κB and NLRP3 increased(P<0.05).Conclusion CUR can increase the antioxidant stress level of keratinocytes,alleviate inflammatory response,promote cell proliferation,and improve cell damage caused by UVB irradiation,which may be related to the inhibition of TLR4/NF-κB/NLRP3 signaling pathway.
10.Research progress in analytical technologies for characterizing pharmacodynamic substance basis of traditional Chinese medicine compound prescriptions
Yuan YUAN ; Di NIU ; Xinxin PANG ; Chaoyang DU ; Yanan WANG ; Ruifang CHEN
China Pharmacy 2025;36(21):2738-2741
Traditional Chinese medicine (TCM) compound prescriptions serve as crucial practical embodiments of TCM theoretical frameworks, characterized by their complex multi-component composition and multi-target interactions. The research on the material basis of their pharmacological effects has gradually become the key to promoting the modernization of TCM. In recent years, new ideas and theories regarding the research on pharmacodynamic substance basis of TCM compound prescriptions have been continuously proposed. This review systematically summarizes and reviews analytical techniques such as targeted fishing technology, spectrum-effect relationship analysis, serum pharmacochemistry, network pharmacology, high-throughput screening, and cell membrane chromatography. It is found that these techniques exhibit unique advantages in areas including target-specific analysis, component-pharmacological effect correlation analysis, identification of the material basis in vitro and in vivo, prediction of multi-target mechanisms, efficient screening of active ingredients, and analysis of interactions between cell membrane receptors. These techniques compensate for the shortcomings of traditional research methods, enhance the systematicness and precision of research on pharmacodynamic substance based TCM compound prescriptions, and can provide theoretical support for the promotion and clinical application of TCM compound prescriptions.

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