1.Treatment of Sepsis-induced Inflammatory Responses with Xijiao Dihuangtang by Modulation of PKM2-mediated One-carbon Metabolism Pathway
Qixiang YAN ; Yeyan ZHU ; Fan GE ; Qimeng SUN ; Leyao YE ; Fang TIAN ; Jun LU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(10):18-26
ObjectiveTo investigate the effects of Xijiao Dihuangtang (XJDHT) on mice with sepsis and cellular models of sepsis and explore its molecular mechanism in alleviating sepsis-induced inflammatory responses via regulating pyruvate kinase M2 (PKM2)-mediated one-carbon metabolism pathway. MethodsForty C57BL/6N mice were randomly divided into four groups: normal group, model group, low-dose XJDHT group (7.7 g·kg-1), and high-dose XJDHT group (15.4 g·kg-1). After one week of continuous gavage, sepsis was induced using cecal ligation and puncture (CLP) in groups except the normal group. 24 h after the surgery, mortality rates in all groups were recorded, and serum cytokines were measured by enzyme linked immunosorbent assay (ELISA). Lung histopathology was examined by hematoxylin-eosin (HE) staining. During the in vitro experiment, the human monocytic leukemia cell line (THP-1) was exposed to various concentrations of XJDHT and treated with lipopolysaccharide (LPS) at a final concentration of 2 mg·L-1 for 24 h. Cell apoptosis was detected using terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay. Protein levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), B-cell lymphoma 2 (Bcl-2), and Bcl-2-associated X protein (Bax) were measured by Western blot. Transcriptome sequencing was performed to analyze differentially expressed genes in all groups and conduct gene ontology (GO) enrichment. Key genes in the one-carbon metabolism pathway, including pyruvate kinase M2 (PKM2), 5-methyltetrahydrofolate-homocysteine methyltransferase (MTR), and phosphoglycerate dehydrogenase (PHGDH), were verified by Western blot. A PKM2 inhibition model was established using shikonin for further protein expression analysis. ResultsAnimal experiments showed that compared with the normal group, the model group exhibited significantly elevated body temperature and lung pathology (P<0.01) and increased serum TNF-α and IL-1β levels (P<0.01). High-dose XJDHT reduced body temperature and lung tissue damage (P<0.01) and significantly decreased serum TNF-α and IL-1β levels (P<0.01). Low-dose XJDHT treatment showed no significant temperature change (P<0.01) but reduced serum TNF-α and IL-1β levels (P<0.01). Transcriptome sequencing and Western blot revealed significant differences in the expression of TNF-α, IL-1β, and one-carbon metabolism genes (PKM2, MTR, and PHGDH) (P<0.01). Cell experiments demonstrated that compared to the normal group, the model group showed elevated protein expressions of TNF-α and IL-1β in THP-1 cells (P<0.01), decreased Bcl-2/Bax ratio, and increased apoptosis (P<0.01). Transcriptome sequencing and Western blot revealed significant differences in the expression of TNF-α, IL-1β, and one-carbon metabolism genes (PKM2, MTR, and PHGDH) (P<0.01). Compared to the model group, high-dose XJDHT significantly increased Bax/Bcl-2 ratio and PHGDH protein expression (P<0.01) and effectively reduced cell apoptosis (P<0.01) while down-regulating protein expressions of TNF-α, IL-1β, PKM2, and MTR (P<0.01). Low-dose XJDHT moderately increased Bax/Bcl-2 ratio and PHGDH protein expression (P<0.05), reduced apoptosis (P<0.05), and decreased IL-1β and MTR protein levels (P<0.05, P<0.01), but there were no significant changes in TNF-α and PKM2 expression. After PKM2 inhibition by shikonin in THP-1 cells, the expression of protein related to one-carbon metabolism was detected. Compared with the blank group, the LPS-induced model group showed significantly upregulated PKM2 and MTR protein expression (P<0.01) and downregulated PHGDH expression (P<0.01). Compared with the model group, shikonin treatment significantly reduced PKM2 expression (P<0.05), increased PHGDH expression (P<0.01), and decreased MTR expression (P<0.05). ConclusionXJDHT can inhibit the release of inflammatory factors in sepsis, and its mechanism is related to the intervention of the PKM2-regulated one-carbon metabolism pathway in macrophages.
2.Mechanism of Pharmacological Liver and Kidney Injuries of Dictamni Cortex Based on UPLC-Q-TOF-MS
Jiahe YAN ; Sujie LIU ; Xiaofan WANG ; Chen WANG ; Jiaxin RUAN ; Fang LU ; Shumin LIU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(20):48-56
ObjectiveThis study aims to reveal the mechanism of liver and kidney injuries caused by Dictamni Cortex and its interrelationship by metabonomics analysis of liver and kidney via ultra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry (UPLC-Q-TOF-MS). MethodsThe content of the marker compounds of Dictamni Cortex was measured by high-performance liquid chromatography (HPLC) to carry out quality control. Sprague Dawley (SD) rats were randomly divided into a blank group (normal saline), an administration group (0.9, 2.7, 8.1 g·kg-1), and a high-dose withdrawal control group, with eight rats in each group. Continuous administration was performed once daily for 28 days. The liver and kidney injuries caused by each administration group were assessed by organ indices, pathological observations, and serum and plasma biochemical indices measured by enzyme-linked immunosorbent assay (ELISA). The potential biomarkers of liver and kidney injuries caused by Dictamni Cortex were screened, and pathway enrichment analysis and correlation analysis were performed based on UPLC-Q-TOF-MS. ResultsCompared with the blank group, both the medium- and low-dose groups showed insignificant damage to the liver and kidney of rats. The high-dose group exhibited the most serious damage, and the level of liver and kidney function indices [alanine aminotransferase (ALT), aspartate aminotransferase (AST), creatinine (Cr), and blood urea nitrogen (BUN)] and serum inflammatory indices ([interleukin 1β (IL-1β), IL-6, and tumor necrosis factor-α (TNF-α)] in the serum were significantly changed (P<0.01). The liver and kidney metabolism pathways and differential metabolites were quite different. Among them, phenylalanine metabolism, niacin and nicotinamide metabolism, and glycerophospholipid metabolism were common pathways. Correlation analysis of differential metabolites showed that there were significant correlations among disorders of 4′-Phosphopantothenoylcysteine, PC (16∶0/15∶0), phenylethylamine, arachidonic acid, and linoleic acid in liver and kidney tissue. ConclusionThe decoction of Dictamni Cortex can cause liver and kidney injuries, and its mechanism may be related to oxidative stress and lipid metabolism disorders. The correlation of differential metabolites indicates the interaction between liver and kidney injuries.
3.Metabolomics Reveals Immune System Domage of Dictamnine
Xiaocan GAI ; Jiaxin RUAN ; Sujie LIU ; Chen WANG ; Xiaofan WANG ; Jiahe YAN ; Yu WANG ; Fang LU ; Shumin LIU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(20):57-65
ObjectiveTo explore the mechanism of the immunotoxicity induced by dictamnine (DIC) in rats and the recovery effect after drug withdrawal by ultra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry, thereby providing a theoretical basis for elucidating the toxic mechanism of DIC. MethodsSD rats were randomized into blank (normal saline), DIC (10 mg·kg-1), and DIC withdrawal (recovery period) groups (n=8). The rats were continuously treated for 7 days, once a day, and the body weight and organ weight were recorded. The levels of interleukin-1 (IL-1), IL-6, and tumor necrosis factor-α (TNF-α) in the serum and immunoglobulin A (IgA), immunoglobulin G (IgG), and immunoglobulin M (IgM) in the spleen were determined by enzyme-linked immunosorbent assay. Hematoxylin-eosin staining was used to observe the pathological changes in the spleen. ultra performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) was employed to screen the potential biomarkers of immune inflammation caused by DIC, and pathway enrichment analysis and correlation analysis were performed. The mRNA levels of IL-1β, TNF-α, lysophosphatidylcholine acyltransferase 2 (LPCAT2), and farnesoid X receptor (FXR) in the serum were determined by Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR). ResultsCompared with the blank group, the DIC group showed elevated levels of IL-1β, IL-6, and TNF-α in the serum (P<0.01), and the DIC withdrawal group showcased lowered levels of IL-1β, IL-6, and TNF-α in the serum (P<0.01). The levels of IgA, IgG, and IgM in the spleen of rats in the DIC group were decreased (P<0.01), while those in the DIC withdrawal group were recovered (P<0.05, P<0.01). Untargeted metabolomics of the serum and spleen screened out 14 common differential metabolites and 14 common metabolic pathways. The Spearman correlation analysis between differential metabolites and inflammatory factors identified PC (32∶0), LysoPC (20∶4/0∶0), LysoPC (P-18∶0/0∶0), taurochenodeoxycholic acid, taurocholic acid, LysoPC [20∶5(5Z,8Z,11Z,14Z,17Z)/0∶0], chenodeoxycholic acid, arachidonic acid, LysoPC (18∶0/0∶0), LysoPC (15∶0/0∶0), LysoPC (16∶0/0∶0), and LysoPC (17∶0/0∶0) as the biomarkers of immunotoxicity induced by DIC in SD rats. In the process of immunotoxicity caused by DIC, lipid metabolism disorders such as glycerophospholipid metabolism, primary bile acid metabolism, and arachidonic acid metabolism were enriched, which was consistent with the DIC-induced inflammatory factors and pathological characteristics of the spleen. Compared with the blank group, the DIC group exhibited up-regulated mRNA levels of IL-1β, TNF-α, LPCAT2, and FXR (P<0.01), and the up-regulation was decreased in the withdrawal group (P<0.01). ConclusionDIC can lead to immune and inflammatory disorders. DIC withdrawal can regulate the expression of biomarkers related to serum and spleen metabolites, regulate the inflammatory metabolic pathway, reduce the inflammation level, and alleviate the metabolic disorders, thus attenuating the potential toxicity induced by DIC.
4.Relationship between Helicobacter pylori infection and disease severity and pathological type of inpatients with intestinal polyps
Wei YOU ; Dalin LU ; Yan CHEN ; Xin WANG ; Yizheng FANG ; Lunshan WANG
Journal of Public Health and Preventive Medicine 2025;36(5):85-88
Objective To investigate the relationship between Helicobacter pylori (Hp) infection and disease severity and pathological type of intestinal polyps in inpatients. Methods The data of 303 inpatients with intestinal polyps in the hospital from August 2022 to February 2025 were collected and analyzed. The clinical characteristics of patients were analyzed, and the influencing factors of pathological types of polyps were explored. Results Among the 303 inpatients with intestinal polyps, there were 135 Hp positive cases and 168 Hp negative cases. The number of polyps, maximum polyp diameter, number of ileocecus/ascending colon/transverse colon polyps, number of descending colon/sigmoid colon/rectal polyps and adenomatous polyps in the Hp-positive group were higher than those in the Hp-negative group (P<0.05). Multivariate logistic regression analysis indicated that age [OR (95%CI)=1.03 (1.01-1.05)] and positive Hp[OR (95%CI)=2.61 (1.62-4.20)] were independent risk factors of occurrence of adenomatous polyps (P<0.05). ROC curve results revealed that the AUCs of age, positive HP and combination in the diagnosis of adenomatous polyps were 0.574, 0.608 and 0.646, and the 95%CI values were 0.509-0.638, 0.545-0.672 and 0.584-0.708 respectively. The efficiency of combination of the two indexes was higher than that of single diagnosis. Conclusion Hp infection is associated with disease severity in inpatients with intestinal polyps, and it may be involved in the occurrence and development of adenomatous polyps.
5.Relationship between Helicobacter pylori infection and disease severity and pathological type of inpatients with intestinal polyps
Wei YOU ; Dalin LU ; Yan CHEN ; Xin WANG ; Yizheng FANG ; Lunshan WANG
Journal of Public Health and Preventive Medicine 2025;36(5):85-88
Objective To investigate the relationship between Helicobacter pylori (Hp) infection and disease severity and pathological type of intestinal polyps in inpatients. Methods The data of 303 inpatients with intestinal polyps in the hospital from August 2022 to February 2025 were collected and analyzed. The clinical characteristics of patients were analyzed, and the influencing factors of pathological types of polyps were explored. Results Among the 303 inpatients with intestinal polyps, there were 135 Hp positive cases and 168 Hp negative cases. The number of polyps, maximum polyp diameter, number of ileocecus/ascending colon/transverse colon polyps, number of descending colon/sigmoid colon/rectal polyps and adenomatous polyps in the Hp-positive group were higher than those in the Hp-negative group (P<0.05). Multivariate logistic regression analysis indicated that age [OR (95%CI)=1.03 (1.01-1.05)] and positive Hp[OR (95%CI)=2.61 (1.62-4.20)] were independent risk factors of occurrence of adenomatous polyps (P<0.05). ROC curve results revealed that the AUCs of age, positive HP and combination in the diagnosis of adenomatous polyps were 0.574, 0.608 and 0.646, and the 95%CI values were 0.509-0.638, 0.545-0.672 and 0.584-0.708 respectively. The efficiency of combination of the two indexes was higher than that of single diagnosis. Conclusion Hp infection is associated with disease severity in inpatients with intestinal polyps, and it may be involved in the occurrence and development of adenomatous polyps.
6.Analyzing the heart-oriented view of the treatment of mental illness in Synopsis of Golden Chamber based on the theory of five-spirit-viscera
ZOU Peng ; Kai CHENG ; Minlong XIA ; Menghan LI ; Shuxian WANG ; Hui KONG ; Yan ZHAO ; Changming ZHAI ; Fang LU
Journal of Beijing University of Traditional Chinese Medicine 2025;48(4):495-500
Mental state is an important part of the normal life activities of the human body, and it is also the most external expression and the most easily obtained information of the physical condition. The normal activities of the mind depend on the normal operation of the viscera, qi, and blood, and are a unified whole that prospers together and suffers together. The theory of the five-spirit-viscera in the Yellow Emperor’s Inner Classic revealed that the normal mental activities of the human body were dominated by the five internal organs, that is, the five internal organs were the body and the five spirits were the function. And it highlighted the viewpoint that the five internal organs store the spirits and are actually one. The heart governs the spirit and belongs to the four internal organs. On this basis, Synopsis of Golden Chamber used the internal organs to diagnose and treat mental diseases, integrating the theory of the five spirits into it, forming a unique method of diagnosis and treatment with the heart as the leading factor and regulating the qi and blood of the four internal organs. It identified the pathogenesis of diseases such as pathogenic crying, lily disease, and hysteria from five levels: heart deficiency and weak qi, heart-lung disharmony, heart-liver disharmony, the heart of the loss of the spleen nourishment, and disharmony between heart and kidney. The treatment was mainly to replenish the deficiency of the viscera and eliminate the pathogens, reflecting the characteristics of regulating the mind and calming the four internal organs. This unique view on diagnosis and treatment has profoundly influenced the diagnosis and treatment theories of mental illnesses by later doctors, and is of great significance to the current clinical treatment of such illnesses.
7.Relationship between metal exposure and blood pressure in rural primary school students in Shiyan City, Hubei Province
YAN Tinghua, FANG Wen, LI Junmin, ZHANG Yao, LU Yu, ZHANG Miao
Chinese Journal of School Health 2024;45(4):581-584
:
To explore the relationship between metal exposure level and blood pressure, so as to provide a scientific basis for verifying the relationship between metal exposure and elevated blood pressure among primary school students.
Methods:
In July 2022, a total of 555 students of second to sixth grade were selected by cluster random sampling method from two primary schools in Zhuxi County, Shiyan City, Hubei Province. A questionnaire survey was conducted to obtain the socio demographic characteristics and living habits of the participants. The height, weight, body mass index(BMI) and blood pressure were obtained by physical examination. At the same time, the urine of the subjects was collected, and the metal mass fraction in urine was detected by inductively coupled plasma mass spectrometry. The relationship between metal mass fraction in urine and blood pressure was analyzed by generalized linear regression.
Results:
The detection rate of elevated blood pressure in primary school students was 15.86% , and there was a statistically significant difference in the detection rate of elevated blood pressure among obese primary school students (yes:37.25%,no:13.69%, χ 2=19.28, P <0.01).There were statistically significant differences in BMI[15.80( 14.69 , 17.92 ),17.87(15.49,20.89)kg/m 2] between the non elevated blood pressure group and the elevated blood pressure group of elementary school students ( Z =-4.67, P <0.01). The geometric mean mass fraction of zinc in urine was the highest ( 6 942.86 μg/g), titanium was the lowest (2.20 μg/g). Zinc and lead were positively correlated with elevated systolic blood pressure( β = 0.054 , 0.014), zinc and cadmium were positively correlated with elevated diastolic blood pressure ( β =0.038,0.029) ( P <0.05).
Conclusions
Metal zinc, lead and cadmium concentration are associated with elevated blood pressure. It is necessary to intervene and control the exposure of zinc, lead and cadmium in the environment to promote the blood pressure health of primary school students.
8.An Enhancer LncRNA Regulates NFE2 Expression and Proliferation in Human Leukemic K562 Cells
Yan-Fei LU ; Song-Ya QU ; Jing-Jing ZHU ; Chao LIU ; Jian WANG ; Bing-She HAN ; Jun-Fang ZHANG
Progress in Biochemistry and Biophysics 2024;51(1):190-201
ObjectiveTranscription factor NFE2 was observed abnormal expression in myeloproliferative neoplasm (MPN) patients. However, how NFE2 is transcriptionally regulated remains ambiguous. This study aims to explore the elements and molecular mechanisms involved in the transcriptional regulation of NFE2. MethodsActive enhancers were predicted by public NGS data and conformed experimentally via dual luciferase reporter assay. After that, PRO-seq and GRO-seq data was used to detect enhancer RNAs transcribed from these enhancers. RACE was utilized to clone the full length enhancer RNA (eRNA) transcripts, and RT-qPCR was used to measure their expression in different leukemia cell lines as well as the transcript levels during induced differentiation. Finally, to investigate the molecular function of the eRNA, overexpression and knockdown of the eRNA via lentivirus system was performed in K562 cells. ResultsWe identified three enhancers regulating NFE2 transcription, which located at -3.6k, -6.2k and +6.3k from NFE2 transcription start site (TSS) respectively. At the -3.6k enhancer, we cloned an eRNA transcript and characterized that as a lncRNA which was expressed and located in the nucleus in three types of leukemia cell lines. When this lncRNA was overexpressed, expression of NFE2 was upregulated and decreases of K562 cell proliferation and migration ability were observed. While knocking down of this lncRNA, the level of NFE2 decreases correspondingly and the proliferation ability of K562 cells increases accordingly. ConclusionWe identified an enhancer lncRNA that regulates NFE2 transcription positively and suppresses K562 cell proliferation.
9.Relationship between school bullying and mental sub-health in middle school students
Dandan LU ; Fang LI ; Huihao WANG ; Yue GUAN ; Quzhen GESANG ; Liang YUAN ; Hong YAN ; Bin YU
Chinese Mental Health Journal 2024;38(2):104-108
Objective:To explore the relationship between school bullying and mental sub-health in middle school students and the potential moderating role of resilience in this relationship.Methods:Totally 792 students aged 10 to 14 years from two middle schools in Wuhan were selected.The Chinese version of Olweus Bully/Victim Questionnaire,Multidimensional Sub-health Questionnaire of Adolescents,and Adolescent Psychological Resilience Scale were used to measure school bullying,mental sub-health,and psychological resilience of students,respectively.Results:Being bullied scores were positively associated with mental sub-health scores(β=1.88).The moderating effect of psychological resilience scores between being the scores of bullied and mental sub-health was statistically significant(β=-0.07).Conclusion:The experience of bullying may be associated with mental sub-health problems of middle school students,and psychological resilience may play a moderating role in the relation-ship between being bullied and mental sub-health.
10.Exploration and Prospect of Quantitative Evaluation of Integrity Risk Prevention and Control in Public Hospitals
Yan CHEN ; Zhuoma JIAHUAN ; Kai WU ; Shiying LI ; Xinyu CUI ; Lu CENG ; Fang ZHU ; Juan XIE
Chinese Hospital Management 2024;44(3):80-83
Objective To explore the quantitative evaluation of integrity risk prevention and control in public hospitals,provide reference for improving the quality and efficiency of integrity risk prevention and control.Methods Self-designed"Inspection Standards for Integrity Risk Prevention and Control of Power Matters in Public Hospitals"was used to score and rate the power matters provided by each functional department/clinical department of West China Hospital of Sichuan University from three aspects:the clarity of power operation process,the accuracy of finding integrity risk points,the effectiveness of prevention and control measures.Results A total of 236 power matters of the hospital were inspected for integrity risk prevention and control,and according to the inspection criteria,57 items were rated as first grade,103 items were rated as second grade,and 76 items were rated as third grade,accounting for 24.15%,43.64%and 32.20%,respectively.The score for the special work of integrity risk prevention and control was 5.82±1.92 points,of which the process dimension score was 2.11±0.75 points,the risk points dimension score was 1.89±0.92 points,the prevention and control dimension score is 1.89± 0.79 points,which reflects the problems of unclear workflow,inaccurate finding of individual risk points,and unspecified prevention and control measures in some units.Conclusion Hospitals should focus on the concreteness,accuracy,salience and quantification in the long-term construction of integrity risk prevention and control from the aspects of thought,behavior,effectiveness and evaluation.


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