1.Establishment of a DNA quantitation method based on DTT-crystal violet
Yuqin YE ; Huang CAO ; Xufeng WU ; Yao TAN ; Quanfu MA
International Journal of Laboratory Medicine 2025;46(13):1575-1580
Objective To establish a DNA quantitation method based on dithiothreitol(DTT)-crystal vio-let.Methods DTT was used to decolorize crystal violet,mixed with different concentrations of λ-DNA and salmon sperm DNA standard samples or concentration standard samples,and the absorbance was read at 595 nm wavelength by microplate reader,and compared with the results of ultraviolet absorbance method.DTT-crystal violet method and ultraviolet absorbance method were used to compare the concentration of plasmid samples and the concentration of genomic DNA samples of cervical exfoliated cells.The protein tolerance of the two methods was assessed by simulating protein contaminants with bovine serum albumin(BSA).Results In the quantification of λ-DNA and salmon sperm DNA,the DTT-crystal violet method had a robust linear correlation between the absorbance at 595 nm and DNA concentration(r2>0.95),and the measured concentrations of the standard samples were not significantly different from the theoretical concentrations of the prepared standard samples(P>0.05).There was no significant difference in the concentration of plasmid samples measured by DTT-crystal violet method and ultraviolet absorption method(P>0.05).The concen-tration of DNA samples from cervical exfoliated cells measured by ultraviolet absorption method was positive-ly correlated with that by DTT-crystal violet method(r=0.94,P<0.01).The concentration of the standard sample containing BSA 1 μg/μL measured by ultraviolet absorption method was higher than that of the con-trol sample,and the difference was statistically significant(P<0.01),whereas the DTT-crystal violet method was not significantly affected(P>0.05).Conclusion DTT-crystal violet method has obvious advantages over the existing DNA quantitation method,and is suitable for DNA quantitative analysis in scientific research and clinic.
2.Application of emergency endoscope in patients suffering from acute gastrointestinal bleeding complicated with acute myocardial infarction: effectiveness, safety, and risk factors
Peng WU ; Yi ZHU ; Zhongman ZHANG ; Huazhong ZHANG ; Xufeng CHEN
Chinese Journal of Emergency Medicine 2025;34(7):959-963
Objective:Acute myocardial infarction (AMI) secondary to acute gastrointestinal bleeding (AGIB) is a common severe condition in emergency department. Currently, there is a lack of sufficient data regarding the use of emergency endoscopy in patients suffering from acute gastrointestinal bleeding (AGIB) complicated with acute myocardial infarction (AMI). The objective of this study is to examine the present status, effectiveness, and safety of emergency endoscopy in such patients, and to determine the factors that influence clinicians' decision-making regarding the use of emergency endoscopy.Methods:Clinical data of AGIB patients complicated with AMI who were admitted to the emergency department of the First Affiliated Hospital of Nanjing Medical University from January 2020 to June 2023 were respectively collected. Based on their survival status at discharge, patients were categorized into survival and non-survival groups. Additionally, according to whether emergency endoscopy was performed, patients were further classified into the emergency endoscopy group and the conventional treatment groupStabilized inverse probability of treatment weighting (SIPTW) and logistic regression analysis were used to explore the correlation between emergency endoscopy and prognosis. A paired wilcoxon test was used to analyze whether emergency endoscopy could worsen myocardial injury. A multiple logistic regression model was used to explore the factors influencing clinicians to make emergency endoscopic decisions.Results:A total of 106 patients with AMI secondary to AGIB were included, and 22 patients underwent emergency endoscopy. After weighing, logistic regression analysis showed that emergency endoscopy was a protective factor for patients' survival after discharge ( OR = 0.808, 95% CI: 0.6755-0.967, P = 0.022). In the emergency endoscopy group, there was no significant change in hypersensitive troponin T before and after emergency endoscopy [ 61.1 (35.4, 164.1) ng/L vs 69.8 (60.1, 159.5) ng/L, P = 0.078]. Previous history of coronary heart disease ( OR = 0.098, 95% CI: 0.015-0.381, P = 0.003) was an independent negative factor for emergency endoscopic decision-making. Glasgow Blatchford Score (GBS) on admission ( OR = 1.217, 95% CI: 1.019-1.482, P = 0.038) was an independent positive factor for emergency endoscopic decision-making. Conclusions:Emergency endoscopy is safe and effective in patients with AMI secondary to AGIB. GBS score and previous history of coronary heart disease were the main factors affecting emergency endoscopic decision-making.
4.Research progress on restarting anticoagulant therapy for anticoagulation related cerebral hemorrhage
Xufeng PAN ; Rong WU ; Haifei CHAI ; Jie WANG ; Xiaoming HU
Journal of Chinese Physician 2024;26(8):1273-1276
With the aging population, the burden of diseases such as atrial fibrillation and venous thrombosis is gradually increasing. Anticoagulant therapy has a positive significance in preventing ischemic stroke, pulmonary embolism, and other related conditions in these patients. However, anticoagulant therapy can have the opposite effect on diseases caused by intracranial hemorrhage, such as falls in the elderly, cerebrovascular accidents, and car accidents. It is still difficult to determine whether and when to restart anticoagulation after cerebral hemorrhage. Although most studies have shown that restarting anticoagulant therapy can reduce stroke risk without significantly increasing bleeding risk, they are mostly based on observational studies, so more high-quality research is needed to guide clinical decision-making. This article reviews the research progress on restart anticoagulation, aiming to provide some assistance for clinical applications.
5.Advances in the treatment of venous thromboembolism in the neurosurgery department
Xufeng PAN ; Rong WU ; Haifei CHAI ; Jie WANG ; Xiaoming HU
Chinese Journal of Primary Medicine and Pharmacy 2024;31(9):1433-1436
Venous thromboembolism is a common comorbidity in neurosurgery department that can lead to life-threatening pulmonary embolism, endangering patient health. The unique characteristics of neurosurgical conditions often present a high risk of bleeding, which complicates the treatment of venous thrombosis. Although numerous observational studies and meta-analyses support the feasibility of initiating early anticoagulation prevention or treatment after hemorrhage stabilization in intracranial hemorrhagic conditions such as traumatic brain injury and cerebral hemorrhage, there is a lack of high-quality clinical research. As a result, neurosurgeons tend to adopt a conservative approach regarding pharmacological prophylaxis and anticoagulant treatment for venous thromboembolism. Key aspects such as the timing of prevention, monitoring, and discontinuation of treatment still require high-quality research to establish definitive guidelines.
6.Progresses of imaging researches for predicting brain age
Yulei ZHANG ; Xufeng YAO ; Tao WU
Chinese Journal of Interventional Imaging and Therapy 2024;21(9):561-564
With the intensify of aging population,accurate assessment of brain health becomes more and more important.Brain age can reflect brain health and cognitive function.Based on different algorithms,modal data and central datasets,imaging is helpful for predicting brain age.The progresses of imaging researches for predicting brain age were reviewed in this article.
7.A consistency comparison between next-generation sequencing and the FISH method for gene rearrangement detection in B-cell lymphomas
Zheng YAN ; Zhihua YAO ; Shuna YAO ; Shuang ZHAO ; Haiying WANG ; Junfeng CHU ; Yuanlin XU ; Jiuyang ZHANG ; Bing WEI ; Jiawen ZHENG ; Qingxin XIA ; Daoyuan WU ; Xufeng LUO ; Wenping ZHOU ; Yanyan LIU
Chinese Journal of Hematology 2024;45(6):561-565
Objective:To compare the consistency of lymphoma multigene detection panels based on next-generation sequencing (NGS) with FISH detection of B-cell lymphoma gene rearrangement.Methods:From January 2019 to May 2023, fusion genes detected by lymphoma-related 413 genes that targeted capture sequencing of 489 B-cell lymphoma tissues embedded in paraffin were collected from Henan Cancer Hospital, and the results were compared with simultaneous FISH detection of four break/fusion genes: BCL2, BCL6, MYC, and CCND1. Consistency was defined as both methods yielding positive or negative results for the same sample. The relationship between fusion mutation abundance in NGS and the positivity rate of cells in FISH was also analyzed.Results:Kappa consistency analysis revealed high consistency between NGS and FISH in detecting the four B-cell lymphoma-related gene rearrangement ( P<0.001 for all) ; however, the detection rates of positive individuals differed for the four genes. Compared with FISH, NGS demonstrated a higher detection rate for BCL2 rearrangement, a lower detection rate for BCL6 and MYC rearrangement, and a similar detection rate for CCND1 rearrangement. No correlation was found between fusion mutation abundance in NGS and the positivity rate of cells in FISH. Conclusions:NGS and FISH detection of B-cell lymphoma gene rearrangement demonstrate overall good consistency. NGS is superior to FISH in detecting BCL2 rearrangement, inferior in detecting MYC rearrangement, and comparable in detecting CCND1 rearrangement.
8.A pilot study of gray matter volume and function changes in patients after cardiopulmonary resuscitation
Gannan WANG ; Qian WU ; Xiaoquan XU ; Xufeng CHEN ; Jinsong ZHANG
Chinese Journal of Emergency Medicine 2023;32(1):32-37
Objective:To explore the structural and functional alterations of related brain regions in patients after cardiopulmonary resuscitation (CPR) by brain magnetic resonance imaging (MRI).Methods:A single-center, observational, cross-sectional study design was used. Patients who had brain MRI scans during hospitalization between July 2020 and July 2021 in Emergency Department of the First Affiliated Hospital of Nanjing Medical University and had good neurologic outcomes were consecutive enrolled in this study. The healthy control (HC) group consisted of age- and sex-matched volunteers. The demographic and clinical data were recorded. The modified Rankin Scale (mRS) was used to check the recovery and degree of continued disabilities when patients performed MRI. Montreal cognitive assessment (MoCA) was used to assess cognitive functions. The analyses of voxel-based morphometry (VBM) and fractional amplitude of low-frequency fluctuation (fALFF) were conducted. After data preprocessing, comparison of gray matter volume (GMV) and fALFF values between the case group and HC group were carried out, and the information of different brain regions was obtained. Partial correlation analyses were performed to evaluate the correlation between the image parameters of different clusters and clinical parameters.Results:Totally 13 patients were enrolled in this study and 13 were in the HC group. All patients achieved good neurologic outcome; mRS was 3 in 1 case, 2 in 3 cases, and 1 in 5 cases during MEI examination. The case group showed significantly lower MoCA score compared with the HC group ( P<0.001). There were significantly decreased GMVs in the right inferior frontal gyrus, superior temporal gyrus, left superior temporal gyrus, and transverse temporal gyrus in the case group. The patients showed significantly decreased fALFF values in the left postcentral gyrus and precentral gyrus, while increased fALFF values in the right putamen than the HC group (voxel-level P<0.001 and cluster-level P<0.05 with GRF correction). In addition, mean fALFF value in the right putamen was negatively correlated with MoCA score in the case group ( r=-0.710, P=0.021). Conclusions:Patients after CPR may have GMVs and neuronal spontaneous activity changes in some brain regions, and VBM and fALFF methods can be used to objectively evaluate the impaired brain functional activity in patients after successful CPR.
9.Effect of 5 methods of critical illness score in the prognosis evaluation of sepsis-associated encephalopathy
Yihao CHEN ; Yuxiong GUO ; Xufeng LI ; Xiaoting YE ; Jingwen ZHANG ; Chun WANG ; Yan HU ; Jing WANG ; Jiaxing WU ; Guilang ZHENG ; Yueyu SUN ; Yiyu DENG ; Yiyun LU
Chinese Journal of Emergency Medicine 2022;31(4):520-527
Objective:To explore the effect of pediatric critical illness score (PCIS), pediatric risk of mortality Ⅲ score (PRISM Ⅲ), pediatric logistic organ dysfunction 2 (PELOD-2), pediatric sequential organ failure assessment (p-SOFA) score and Glasglow coma scale (GCS) in the prognosis evaluation of septic-associated encephalopathy (SAE).Methods:The data of children with SAE admitted to the Pediatric Intensive Care Unit (PICU), Guangdong Provincial People’s Hospital, Guangdong Academy of Medical Sciences from January 2010 to December 2020 were retrospectively analyzed. They were divided into the survival and death groups according to the clinical outcome on the 28th day after admission. The efficiency of PCIS, PRISM Ⅲ, PELOD-2, p-SOFA and GCS scores for predicting death were evaluated by the area under the ROC curve (AUC). The Hosmer-Lemeshow goodness-of-fit test assessed the calibration of each scoring system.Results:Up to 28 d after admission, 72 of 82 children with SAE survived and 10 died, with a mortality rate of 12.20%. Compared with the survival group, the death group had significantly lower GCS [7 (3, 12) vs. 12 (8, 14)] and PCIS scores [76 (64, 82) vs. 82 (78, 88)], and significantly higher PRISM Ⅲ [14 (12, 17) vs. 7 (3, 12)], PELOD-2 [8 (5, 13) vs. 4 (2, 7)] and p-SOFA scores [11 (5, 12) vs. 6 (3, 9)] ( P<0.05). The AUCs of PCIS, PRISM Ⅲ, PELOD-2, p-SOFA and GCS scores for predicting SAE prognosis were 0.773 ( P=0.012, AUC>0.7), 0.832 ( P=0.02, AUC>0.7), 0.767 ( P=0.014, AUC>0.7), 0.688 ( P=0.084, AUC<0.7), and 0.692 ( P=0.077,AUC<0.7), respectively. Hosmer-Lemeshow goodness-of-fit test showed that PCIS ( χ2=5.329, P=0.722) predicted the mortality and the actual mortality in the best fitting effect, while PRISM Ⅲ ( χ2=12.877, P=0.177), PELOD-2 ( χ2=8.487, P=0.205), p-SOFA ( χ2=9.048, P=0.338) and GCS ( χ2=3.780, P=0.848) had poor fitting effect. Conclusions:The PCIS, PRISM Ⅲ and PELOD-2 scores have good predictive ability assessing the prognosis of children with SAE, while the PCIS score can more accurately evaluate the fitting effect of SAE prognosis prediction.
10.Improvement of synthetic route of brivudine
Xufeng WU ; Shiying CHAI ; Jianhong LIU ; Zhihui WANG ; Ruifeng YIN ; Ruiwei CAO ; Xinqiang SUN
Journal of China Pharmaceutical University 2022;53(1):41-45
This article summarizes and analyzes the reported synthetic routes of brivudine in patent and literature.2′-Deoxyuridine was employed as starting material, affording brivudine through iodization, heck coupling, hydrolysis, decarboxylation, bromination and recrystallization.After optimization of reaction conditions of each step, a synthetic route suitable for industrial production was achieved with simple synthetic process, high yield and excellent purity.

Result Analysis
Print
Save
E-mail