1.Differences in deltamethrin resistance and kdr gene mutation in Culex tritaeniorhynchus population in and outside the Yellow Sea wetland
Xiao-er ZHANG ; Zhi-ming WU ; Ye TIAN ; Qian CUI ; Yu-qian JI ; Huan WANG ; Shu-juan YANG ; Yi-chao ZHAO ; Yu WANG ; Hua-yu YIN ; Yu DING ; Guo-jin YAN ; Min-sen ZHAO ; Shou-gang ZHANG ; Bing-dong SONG ; Hong-na CHEN ; Jian GAO ; Wei-fang YANG ; Yu-fu ZHANG ; Hui LIU ; Hong-liang CHU
Acta Parasitologica et Medica Entomologica Sinica 2026;33(2):101-107
Objective To gain insights into the biological characteristics of different populations of Culex tritaeniorhynchus within and around the Yellow Sea wetland from the perspective of the occurrence of resistance, we investigated the levels of resistance to deltamethrin and kdr gene mutation in the wetland and its peripheral areas. Methods Specimens were collected from Cx. tritaeniorhynchus populations at two monitoring sites in the Rare Bird National Nature Reserve and Tiaozi Ni Wetland Scenic Area, and also from two populations in Yancheng City and the Liuhe District of Nanjing, and the resistance of these mosquitoes to deltamethrin was determined using the CDC biotest bottle method. For each concentration of deltamethrin assessed, a random subset of exposed specimens was selected for amplification of the kdr gene fragment, followed by Sanger sequencing to identify and analyze resistance-associated mutations. Results The LC50 levels of deltamethrin among mosquitoes from the four populations in Luhe, Yancheng, the Rare Bird National Nature Reserve and the Tiaozi Ni Wetland Scenic Area were 2.048 5, 7.798 2, 3.473 3, and 17.695 5 mg/mL, respectively, with corresponding concentrations of deltamethrin ranging from 0.005 to 5.000,0.050 to 50.000,0.050 to 25.000 and 0.050 to 50.000 mg/mL, respectively. Furthermore, the ranges of the KT50 values were 11.76-107.43, 67.05-216.30,29.77-107.43 and 28.40-329.51 min; the 1-h knockdown rates were 34.58%-99.15%, 9.52%-43.80%, 55.09%-73.01%, and 10.09%-68.07%; and the 24-h mortality rates were 12.15%-67.52%,9.52%-79.56%,13.17%-82.21%, and 11.01%-78.99%, respectively. With respect to kdr gene mutation, we assayed a total of 63,70,59, and 57 mosquitoes for the four populations, for which we detected L1014F mutation frequencies of 14.29%, 35.00%, 20.34%, and 31.58%, respectively, with a majority of these mutations being heterozygous for resistance. In addition, five adult mosquitoes were identified has having synonymous mutations at site 1011[i. e. , AAT(asparagine)mutation to AAC(asparagine)]. Conclusions Our findings revealed the clear resistance of Cx. tritaeniorhynchus to deltamethrin in the Yancheng region of the Yellow Sea wetland, and the resistance phenotype and kdr frequency of Cx. tritaeniorhynchus in the wetland environment were comparable to those of Cx. tritaeniorhynchus in the wetland environment, thereby indicating that the resistance of different populations of Cx. tritaeniorhynchus was homogeneous under the pressure of different insecticide selection within and around the wetland. However, the underlying mechanisms need to be further studied.
2.Mosquito surveillance and pathogen detection analysis in Jiangsu Province from 2020 to 2024
Jian GAO ; Hong-na CHEN ; Zhi-ming WU ; Wei-fang YANG ; Hong-liang CHU
Acta Parasitologica et Medica Entomologica Sinica 2026;33(3):187-191,203
Objective To analyze mosquito species composition and prevalence of pathogens carried by mosquitoes in Jiangsu Province from 2020 to 2024 and to provide a scientific basis for the prevention and control of mosquito-borne diseases. Methods Mosquitoes were collected using CO2-baited traps. The presence of mosquito-borne pathogens including dengue virus, Japanese encephalitis virus, yellow fever virus, West Nile virus, Chikungunya virus, and Sindbis virus was tested using commercial kits for fluorescence-based quantitative PCR. Results The most prevalent mosquito species found across different regions of Jiangsu Province were Culex pipiens pallens and Cx. tritaeniorhynchus, accounting for 81.35% to 93.01% of the total, followed by Armigeres subalbatus and Anopheles sinensis, which comprised 5.89% to 15.85% of the total. The densities of other monitored mosquito species were relatively low. Pathogen detection revealed that, only Cx. tritaeniorhynchus tested positive for Japanese encephalitis virus nucleic acid, whereas the remaining four mosquito species tested negative for the nucleic acids of all other six pathogens. The overall positivity rate of Japanese encephalitis virus (JEV) in Cx. tritaeniorhynchus showed an initially increasing trend followed by a decrease and a subsequent increase. The peak detection rate occurred in 2021 (11.93%), with a much lower positive rate of 0.80% in 2023. Genotyping result revealed that all JEV strains in Jiangsu Province belonged to the G1 genotype and were genetically closest to the strain Shangdong08DY.29 (MN072875.1). Conclusions The incidence of Japanese encephalitis virus carried by mosquito, especially Cx. tritaeniorhynchus, in Jiangsu Province shows an upward trend, requiring sustained surveillance. A variety of monitoring method should be adopted to survey mosquitoes across different habitats, ensuring the accuracy and comprehensiveness of the monitoring result.
3.Investigations of tick habitats and potential pathogens carried in urban environments of Jiangsu Province
Zhi-ming WU ; Ye TIAN ; Wei-fang YANG ; Hong-na CHEN ; Jian GAO ; Hui LIU ; Yu-fu ZHANG ; Gan CAO ; Hong-liang CHU
Acta Parasitologica et Medica Entomologica Sinica 2026;33(3):192-197
Objective To investigate the species composition, distribution, density, and pathogen-carrying status of host-seeking ticks in urban environments of Jiangsu Province, and to provide a scientific basis for the prevention and control of tick-borne diseases in urban settings. Methods From April to September 2023, a survey on host-seeking ticks was conducted in three types of habitats (city parks, residential areas, and street greenbelts) across 54 counties (cities, districts) of 13 prefecture-level cities in Jiangsu Province by using the flagging method. RStudio 4.4.3 (Windows version) was employed to perform chi-square test analysis on the species composition of ticks across different months, and Friedman test (for multiple paired samples) was applied to analyze monthly changes in the average tick density index among different habitats. Quantitative real-time PCR was performed to detect pathogens in the captured ticks. Results A total of 863 host-seeking ticks were captured from 54 habitats in 24 counties (cities, districts) of 11 prefecture-level cities (Nanjing, Wuxi, Changzhou, Suzhou, Nantong, Huai'an, Yancheng, Yangzhou, Zhenjiang, Taizhou, and Suqian) in Jiangsu Province, whereas no ticks were captured in Xuzhou and Lianyungang. The collected ticks belonged to three species, with Haemaphysalis longicornis being the dominant species (accounting for 82.62%), followed by Haemaphysalis flava and Hae. phasiana. The average density index of ticks in positive habitats was 3.20 ticks/ (flag·100 m). In terms of tick density among prefecture-level cities, Wuxi had the highest density[13.55 ticks/(flag·100 m)], whereas Nanjing and Taizhou had the lowest[both,0.20 ticks/(flag·100 m)]. With respect to the types of positive habitats,46 positive city parks,7 positive residential areas, and 1 positive street greenbelt were found, with the average tick density index being 3.46, 1.86, and 3.60 ticks/flag · 100 m, respectively. No statistically significant differences were observed in the monthly variations in the average tick density index among different habitats (P>0.05). Pathogen detection showed no Bandavirus dabieense, Rickettsia, or Borrelia burgdorferi in the collected ticks. Conclusions Host-seeking ticks have established populations in urban environments across most prefecture-level cities in Jiangsu Province, with Hae. longicornis identified as the dominant species. City parks serve as the primary breeding habitats for these ticks. Strengthening tick surveillance and control measures in densely populated and frequently visited areas, such as city parks and residential neighborhoods, is crucial for the effective prevention and control of urban tick-borne diseases.
4.Expert consensus on management of instrument separation in root canal therapy.
Yi FAN ; Yuan GAO ; Xiangzhu WANG ; Bing FAN ; Zhi CHEN ; Qing YU ; Ming XUE ; Xiaoyan WANG ; Zhengwei HUANG ; Deqin YANG ; Zhengmei LIN ; Yihuai PAN ; Jin ZHAO ; Jinhua YU ; Zhuo CHEN ; Sijing XIE ; He YUAN ; Kehua QUE ; Shuang PAN ; Xiaojing HUANG ; Jun LUO ; Xiuping MENG ; Jin ZHANG ; Yi DU ; Lei ZHANG ; Hong LI ; Wenxia CHEN ; Jiayuan WU ; Xin XU ; Jing ZOU ; Jiyao LI ; Dingming HUANG ; Lei CHENG ; Tiemei WANG ; Benxiang HOU ; Xuedong ZHOU
International Journal of Oral Science 2025;17(1):46-46
Instrument separation is a critical complication during root canal therapy, impacting treatment success and long-term tooth preservation. The etiology of instrument separation is multifactorial, involving the intricate anatomy of the root canal system, instrument-related factors, and instrumentation techniques. Instrument separation can hinder thorough cleaning, shaping, and obturation of the root canal, posing challenges to successful treatment outcomes. Although retrieval of separated instrument is often feasible, it carries risks including perforation, excessive removal of tooth structure and root fractures. Effective management of separated instruments requires a comprehensive understanding of the contributing factors, meticulous preoperative assessment, and precise evaluation of the retrieval difficulty. The application of appropriate retrieval techniques is essential to minimize complications and optimize clinical outcomes. The current manuscript provides a framework for understanding the causes, risk factors, and clinical management principles of instrument separation. By integrating effective strategies, endodontists can enhance decision-making, improve endodontic treatment success and ensure the preservation of natural dentition.
Humans
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Root Canal Therapy/adverse effects*
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Consensus
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Root Canal Preparation/adverse effects*
5.Clinical practice guidelines for the diagnosis and treatment of atopic dermatitis with integrative traditional Chinese and Western medicine.
Xin-Ran DU ; Meng-Yi WU ; Mao-Can TAO ; Ying LIN ; Chao-Ying GU ; Min-Feng WU ; Yi CAO ; Da-Can CHEN ; Wei LI ; Hong-Wei WANG ; Ying WANG ; Yi WANG ; Han-Zhi LU ; Xin LIU ; Xiang-Fei SU ; Fu-Lun LI
Journal of Integrative Medicine 2025;23(6):641-653
Traditional Chinese medicine (TCM) is a well-accepted therapy for atopic dermatitis (AD). However, there are currently no evidence-based guidelines integrating TCM and Western medicine for the treatment of AD, limiting the clinical application of such combined approaches. Therefore, the China Association of Chinese Medicine initiated the development of the current guideline, focusing on key issues related to the use of TCM in the treatment of AD. This guideline was developed in accordance with the principles of the guideline formulation manual published by the World Health Organization. A comprehensive review of the literature on the combined use of TCM and Western medicine to treat AD was conducted. The findings were extensively discussed by experts in dermatology and pharmacy with expertise in both TCM and Western medicine. This guideline comprises 23 recommendations across seven major areas, including TCM syndrome differentiation and classification of AD, principles and application scenarios of TCM combined with Western medicine for treating AD, outcome indicators for evaluating clinical efficacy of AD treatment, integration of TCM pattern classification and Western medicine across disease stages, daily management of AD, the use of internal TCM therapies and proprietary Chinese medicines, and TCM external treatments. Please cite this article as: Du XR, Wu MY, Tao MC, Lin Y, Gu CY, Wu MF, Cao Y, Chen DC, Li W, Wang HW, Wang Y, Wang Y, Lu HZ, Liu X, Su XF, Li FL. Clinical practice guidelines for the diagnosis and treatment of atopic dermatitis with integrative traditional Chinese and Western medicine. J Integr Med. 2025; 23(6):641-653.
Dermatitis, Atopic/drug therapy*
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Humans
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Medicine, Chinese Traditional/methods*
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Integrative Medicine
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Drugs, Chinese Herbal/therapeutic use*
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Practice Guidelines as Topic
6.Bear Bile Powder Ameliorates LPS-Induced Acute Lung Injury by Inhibiting CD14 Pathway and Improving Intestinal Flora: Exploration of "Fei (Lung)-Dachang (Large Intestine) Interaction" Theory.
Long CHENG ; Hui-Ling TIAN ; Hong-Yuan LEI ; Ying-Zhou WANG ; Ma-Jing JIAO ; Yun-Hui LIANG ; Zhi-Zheng WU ; Xu-Kun DENG ; Yong-Shen REN
Chinese journal of integrative medicine 2025;31(9):821-829
OBJECTIVE:
To explore the effect of bear bile powder (BBP) on acute lung injury (ALI) and the underlying mechanism.
METHODS:
The chemical constituents of BBP were analyzed by ultra-high-pressure liquid chromatography-mass spectrometry (UPLC-MS). After 7 days of adaptive feeding, 50 mice were randomly divided into 5 groups by a random number table (n=10): normal control (NC), lipopolysaccharide (LPS), dexamethasone (Dex), low-, and high-dose BBP groups. The dosing cycle was 9 days. On the 12th and 14th days, 20 µL of Staphylococcus aureus solution (bacterial concentration of 1 × 10-7 CFU/mL) was given by nasal drip after 1 h of intragastric administration, and the mice in the NC group was given the same dose of phosphated buffered saline (PBS) solution. On the 16th day, after 1 h intragastric administration, 100 µL of LPS solution (1 mg/mL) was given by tracheal intubation, and the same dose of PBS solution was given to the NC group. Lung tissue was obtained to measure the myeloperoxidase (MPO) activity, the lung wet/dry weight ratio and expressions of CD14 and other related proteins. The lower lobe of the right lung was obtained for pathological examination. The concentrations of inflammatory cytokines including interleukin (IL)-6, tumour necrosis factor α (TNF-α ) and IL-1β in the bronchoalveolar lavage fluid (BALF) were detected by enzyme linked immunosorbent assay, and the number of neutrophils was counted. The colonic contents of the mice were analyzed by 16 sRNA technique and the contents of short-chain fatty acids (SCFAs) were measured by gas chromatograph-mass spectrometer (GC-MS).
RESULTS:
UPLC-MS revealed that the chemical components of BBP samples were mainly tauroursodeoxycholic acid and taurochenodeoxycholic acid sodium salt. BBP reduced the activity of MPO, concentrations of inflammatory cytokines, and inhibited the expression of CD14 protein, thus suppressing the activation of NF-κB pathway (P<0.05). The lung histopathological results indicated that BBP significantly reduced the degree of neutrophil infiltration, cell shedding, necrosis, and alveolar cavity depression. Moreover, BBP effectively regulated the composition of the intestinal microflora and increased the production of SCFAs, which contributed to its treatment effect (P<0.05).
CONCLUSIONS
BBP alleviates lung injury in ALI mouse through inhibiting activation of NF-κB pathway and decreasing expression of CD14 protein. BBP may promote recovery of ALI by improving the structure of intestinal flora and enhancing metabolic function of intestinal flora.
Animals
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Acute Lung Injury/pathology*
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Lipopolysaccharides
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Ursidae
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Gastrointestinal Microbiome/drug effects*
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Bile/chemistry*
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Lipopolysaccharide Receptors/metabolism*
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Powders
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Male
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Lung/drug effects*
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Mice
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Peroxidase/metabolism*
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Signal Transduction/drug effects*
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Cytokines/metabolism*
7.Research Progress of Vagal Nerve Regulation Mechanism in Acupuncture Treatment of Atrial Fibrillation.
Lu-Lu CAO ; Hui-Rong LIU ; Ya-Jie JI ; Yin-Tao ZHANG ; Bing-Quan WANG ; Xiao-Hong XUE ; Pei WANG ; Zhi-Hui LUO ; Huan-Gan WU
Chinese journal of integrative medicine 2025;31(3):281-288
Atrial fibrillation (AF) is the most common arrhythmia in clinical practice. It has a high prevalence and poor prognosis. The application of antiarrhythmic drugs and even surgery cannot completely treat the disease, and there are many sequelae. AF can be classified into the category of "palpitation" in Chinese medicine according to its symptoms. Acupuncture has a significant effect on AF. The authors find that an important mechanism of acupuncture in AF treatment is to regulate the cardiac vagus nerve. Therefore, this article intends to review the distribution and function of vagus nerve in the heart, the application and the regulatroy effect for the treatment of AF.
Atrial Fibrillation/physiopathology*
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Humans
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Acupuncture Therapy
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Vagus Nerve/physiology*
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Animals
8.Comparative study on the predictive abilities of multiple inflammatory markers for the poor outcome of intracerebral hemorrhage
Qin HUANG ; Ziwei SONG ; Hongla KUANG ; Zhi ZHANG ; Yuping ZHU ; Lin WU ; Daojun HONG ; Jing LIN ; Min ZHU
Chinese Journal of Nervous and Mental Diseases 2025;51(2):103-108
Objective This study aimed to investigate the relationship between multiple inflammatory markers and poor outcome in patients with intracerebral hemorrhage,and to compare their predictive abilities.Methods We retrospectively analyzed the patients with intracerebral hemorrhage admitted to the Department of Neurology of the First Affiliated Hospital of Nanchang University from January 1,2015 to March 31,2023.According to the Modified Rankin Scale at 90 days after onset,the patients were divided into good outcome(mRS≤2 points)and poor outcome(mRS score≥3 points).Clinical information,laboratory examinations and follow-up data were compared between the two groups.Inflammatory markers include neutrophil to lymphocyte ratio(NLR),platelet to lymphocyte ratio(PLR),monocyte to high-density lipoprotein ratio(MHR),systemic inflammatory response index(SIRI),systemic immune-inflammation index(SII),white blood cell count to mean platelet volume ratio(WMR),lymphocyte to monocyte ratio(LMR),and systemic coagulation-inflammation index(SCI).Univariate and multivariate logistic regression analysis were used to analyze the predictors of poor prognosis after intracerebral hemorrhage,and receiver operating characteristic curve(ROC)was constructed to compare their predictive ability.Results A total of 510 patients with intracerebral hemorrhage were included.Of those,297(58.2%)had good outcome,and 213(41.8%)had poor outcome.Comparison of baseline characteristics demonstrated that patients with poor prognosis had higher levels of white blood cells,neutrophils,high-density lipoprotein,fibrinogen,NLR,PLR,SIRI,SII,WMR at admission,larger baseline hematoma volume and baseline perihematomal edema volume,a higher proportion of lobar hemorrhage,older age,and lower levels of platelets,lymphocytes,LMR,and SCI(P<0.05).Multivariate regression analysis showed that NLR(OR:1.081,95%CI:1.032~1.1131,P=0.001),SIRI(OR:1.089,95%CI:1.014~1.169,P=0.019),SII(OR:1.000,95%CI:1.000~1.001,P=0.011),WMR(OR:2.627,95%CI:1.267~5.445,P=0.009)were independently associated with poor prognosis in patients with ICH.In ROC analysis,the area under the curve of NLR(0.729,95%CI:0.685~0.774)was higher than SIRI(0.692,95%CI:0.645~0.738),SII(0.688,95%CI:0.641~0.735)and WMR(0.65,95%CI:0.602~0.698)for predicting poor outcomes.Conclusion NLR,SIRI,SII and WMR at admission are independently associated with poor outcomes in patients with intracerebral hemorrhage,and NLR has the strongest predictive ability.
9.Development and validation of a machine learning-based explainable prediction model for the outcome of patients with spontaneous intracerebral hemorrhage
Hong YUE ; Zhi GENG ; Zhaoping YU ; Chi ZHANG ; Xuechun LIU ; Juncang WU ; Aimei WU
International Journal of Cerebrovascular Diseases 2025;33(6):420-428
Objectives:To evaluate the predictive value of Tabular Prior-data Fitted Network(TabPFN) for short-term outcome in patients with spontaneous intracerebral hemorrhage (sICH), and compared with the Extreme Gradient Boosting (XGboost) model and traditional logistic regression (LR) model. Methods:Patients with sICH admitted to the Department of Neurology, Hefei Second People's Hospital from January 2018 to March 2024 were included retrospectively. The demographic and baseline data were collected. At 3 months after onset, the modified Rankin Scale score was used to determine the outcome, 0-2 was defined as good outcome and >2 was defined as poor outcome. All enrolled patients were randomly divided into a training set and a testing set at a ratio of 7:3. Feature selection was performed using recursive feature elimination (RFE) method, and then the selected feature variables were included into TabPFN, XGboost, and LR models for training and testing. The area under the curve (AUC) of receiver operating characteristic (ROC) curve was used to evaluate the predictive ability of the models. Shapley additive explanations (SHAP) method was used for model interpretation.Results:A total of 547 patients with sICH were enrolled, including 367 males (67.1%), with a median age of 65 (interquartile range, 54-76) years. Two hundred twenty-six patients (41.3%) had poor outcome. Age, baseline blood pressure (systolic blood pressure, diastolic blood pressure), baseline laboratory tests (white blood cell count, red blood cell count, platelet count, neutrophil count, hemoglobin, fasting blood glucose, creatinine, uric acid, urea nitrogen, alanine aminotransferase, aspartate aminotransferase), hematoma rupture into the ventricle, island sign, baseline hematoma volume, and baseline National Institutes of Health Stroke Scale (NIHSS) score were selected as characteristic variables using RFE method. ROC curve analysis showed that the ROC AUC for TabPFN, Xgboost, and LR models predicting poor short-term outcome in the testing set were 0.918 (95% confidence interval [ CI] 0.870-0.966], 0.883 (95% CI 0.826-0.940), and 0.905 (95% CI 0.854-0.957), respectively. SHAP analysis showed that the top four important variables in the TabPFN model were baseline NIHSS score, baseline hematoma volume, baseline aspartate aminotransferase, and age. Conclusions:The TabPFN model is superior to the LR model and the XGBoost model in predicting poor outcome in patients with sICH. In the TabPFN model, baseline NIHSS score, baseline hematoma volume, aspartate aminotransferase, and age are the most important predictors of poor outcome in patients with sICH.Objectives To evaluate the predictive value of Tabular Prior-data Fitted Network(TabPFN) for short-term outcome in patients with spontaneous intracerebral hemorrhage (sICH), and compared with the Extreme Gradient Boosting (XGboost) model and traditional logistic regression (LR) model. Methods Patients with sICH admitted to the Department of Neurology, Hefei Second People's Hospital from January 2018 to March 2024 were included retrospectively. The demographic and baseline data were collected. At 3 months after onset, the modified Rankin Scale score was used to determine the outcome, 0-2 was defined as good outcome and >2 was defined as poor outcome. All enrolled patients were randomly divided into a training set and a testing set at a ratio of 7:3. Feature selection was performed using recursive feature elimination (RFE) method, and then the selected feature variables were included into TabPFN, XGboost, and LR models for training and testing. The area under the curve (AUC) of receiver operating characteristic (ROC) curve was used to evaluate the predictive ability of the models. Shapley additive explanations (SHAP) method was used for model interpretation. Results A total of 547 patients with sICH were enrolled, including 367 males (67.1%), with a median age of 65 (interquartile range, 54-76) years. Two hundred twenty-six patients (41.3%) had poor outcome. Age, baseline blood pressure (systolic blood pressure, diastolic blood pressure), baseline laboratory tests (white blood cell count, red blood cell count, platelet count, neutrophil count, hemoglobin, fasting blood glucose, creatinine, uric acid, urea nitrogen, alanine aminotransferase, aspartate aminotransferase), hematoma rupture into the ventricle, island sign, baseline hematoma volume, and baseline National Institutes of Health Stroke Scale (NIHSS) score were selected as characteristic variables using RFE method. ROC curve analysis showed that the ROC AUC for TabPFN, Xgboost, and LR models predicting poor short-term outcome in the testing set were 0.918 (95% confidence interval [ CI] 0.870-0.966], 0.883 (95% CI 0.826-0.940), and 0.905 (95% CI 0.854-0.957), respectively. SHAP analysis showed that the top four important variables in the TabPFN model were baseline NIHSS score, baseline hematoma volume, baseline aspartate aminotransferase, and age. Conclusions The TabPFN model is superior to the LR model and the XGBoost model in predicting poor outcome in patients with sICH. In the TabPFN model, baseline NIHSS score, baseline hematoma volume, aspartate aminotransferase, and age are the most important predictors of poor outcome in patients with sICH.
10.Mechanism of Trifolin in Attenuating Hypertension-Induced Renal Cell Apoptosis via Modulation of the MAPK Signaling Pathway
Meizhu WU ; Zhi GUO ; Yi XIE ; Hongshu LIU ; Hong CHEN ; Xinbiao LIN ; Rongji CHEN ; Aling SHEN ; Jun PENG
Journal of Sichuan University (Medical Sciences) 2025;56(5):1273-1280
Objective To investigate the potential therapeutic effects of trifolin on hypertension-induced renal injury,as well as the key targets and pathways involved.Methods The mRNA transcriptional profiles of peripheral blood clinical samples from hypertensive patients were analyzed using Gene Expression Omnibus(GEO),a high-throughput gene expression database.The network pharmacology method was employed to screen key targets of trifolin in treating hypertension-induced renal injury.Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analyses were conducted.NRK-52E cells,a rat renal proximal tubular cell line,were used to construct an angiotensin Ⅱ(Ang Ⅱ)-stimulated cell model.Flow cytometry was performed to assess cell apoptosis rates and Western blotting was performed to determine the expression levels of apoptosis-related proteins,including Bax,Bcl-2,cleaved caspase-3,and caspase-3,and the phosphorylation and total protein levels of the key MAPK pathway proteins,including ERK,p38 MAPK,and JNK.Results Analysis of the dataset GSE75360 revealed that,compared with healthy controls,3 331 genes were upregulated and 3 197 genes were downregulated in peripheral blood mononuclear cells of hypertensive patients.According to network pharmacology analysis,472 potential targets of trifolin were identified,including CASP3 and MAPK1.Protein-protein interaction network analysis showed that these targets were closely associated with apoptosis regulatory signaling pathways.GO and KEGG pathway enrichment analyses indicated that trifolin was significantly enriched in pathways associated with negative regulation of apoptosis,apoptotic signaling pathways,and the MAPK signaling pathway.The in vitro experiments confirmed that,compared with the Ang Ⅱ group,trifolin intervention inhibited apoptosis in Ang Ⅱ-stimulated NRK-52E cells,suppressed the expression of Bax and cleaved caspase-3,promoted Bcl-2 expression,and inhibited the phosphorylation of p38 MAPK,ERK,and JNK(P<0.05).Conclusion Trifolin may exert its protective effect against hypertension-induced renal injury by inhibiting Ang Ⅱ-induced NRK-52E cell apoptosis and regulating the MAPK signaling pathway,representing an important mechanism underlying its therapeutic action.


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