1.Effect of Huanglian Jiedutang in Regulating Ferroptosis in Mice with Atherosclerosis Based on Nrf2/GPX4 Signaling Pathway
Zhaohui GONG ; Li GAO ; Huiqi ZHAI ; Jinzi YU ; Qingmin CHU ; Chuanjin LUO ; Lijin QING ; Wei WU ; Rong LI
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(3):22-28
ObjectiveTo study the mechanism of Huanglian Jiedutang (HLJDT) in treating mice with atherosclerosis (AS) by improving ferroptosis. MethodsA total of 10 SPF C57BL/6J mice were selected as a normal group, and 50 ApoE-/- mice were randomly divided into five groups: model group, low-dose group of HLJDT, medium-dose group of HLJDT, high-dose group of HLJDT, and atorvastatin (ATV) group. ApoE-/- mice were fed a high-fat diet for eight weeks to establish the AS model, and at the 9th week, they were given normal saline, low, medium, and high doses of HLJDT (3.9, 7.8, 15.6 g·kg-1·d-1), and atorvastatin calcium tablets (0.01 g·kg-1·d-1), respectively, for a total of eight weeks. The formation of aortic plaque in mice was observed by gross oil red O staining and Masson staining. The levels of total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), triglyceride (TG), and high-density lipoprotein cholesterol (HDL-C) in blood fat were measured by the automatic biochemical analyzer, and the mitochondrial structure of the aorta was observed by transmission electron microscopy. The content of serum superoxide dismutase (SOD) in serum was detected by enzyme-linked immunosorbent assay (ELISA). The content of reduced glutathione (GSH) in serum was detected by the microplate method, and that of malondialdehyde (MDA) in serum was detected by the TBA method. The protein expression of nuclear factor E2-associated factor 2 (Nrf2)/glutathione peroxidase 4 (GPX4) signaling pathway was detected by Western blot. ResultsCompared with those of the normal group, the contents of TC, LDL-C, TG, HDL-C, and MDA in the serum and the aortic vascular plaque deposition of the model group were significantly increased (P<0.01), while the expression levels of SOD and GSH in serum, as well as Nrf2, solute carrier family 7 member 11 (SLC7A11), and GPX4 in aorta were significantly decreased (P<0.01). Mice in the model group appeared mitochondrial fragmentation and vacuolation in the aorta, volume atrophy, mitochondrial crista reduction, or a loose and disorganized form. Compared with those in the model group, the aortic vascular plaque deposition was significantly decreased in the low-dose, medium-dose, and high-dose groups of HLJDT and ATV group, and the contents of serum TC, LDL-C, TG, and MDA in serum were significantly decreased (P<0.05, P<0.01). The contents of serum SOD and GSH and the expression levels of Nrf2, SLC7A11, and GPX4 in the aorta were increased (P<0.05, P<0.01), and the symptoms of aortic mitochondrial vacuolation were alleviated. The number of cristae was increased, and they were ordered neatly. ConclusionHLJDT can reduce aortic vascular plaque deposition, decrease blood lipid and MDA expression, increase SOD and GSH expression, and ameliorate the pathological changes of ferroptosis, the mechanism of which is related to the Nrf2/GPX4 signaling pathway.
2.Effect of Qingxin Jieyu Granules on Artery Thrombosis and Akt/NF-κB Signaling Pathway in EA.hy926 Cells Exposed to TNF-α
Chenchen HE ; Chenyi WEI ; Zhenghao LYU ; Qiaoyan CAI ; Zhuye GAO ; Ling ZHANG ; Jianfeng CHU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(3):89-97
ObjectiveTo observe the effects of Qingxin Jieyu granules (QXJYG) on FeCl3-induced carotid artery thrombosis in rats and on the expression of thrombosis-related proteins tissue factor (TF) and tissue factor pathway inhibitor (TFPI) as well as the protein kinase B (Akt)/nuclear factor-κB (NF-κB) signaling pathway in EA.hy926 cells exposed to tumor necrosis factor-α (TNF-α), thus preliminarily exploring the mechanism of QXJYG in inhibiting thrombosis. MethodsThirty-six SD rats were randomized into normal control, model, positive control (aspirin, 9 mg·kg-1), and low-, medium-, and high-dose (0.99, 1.98, 3.96 g·kg-1, respectively) QXJYG groups (n=6). The rats in the drug treatment groups were administrated with corresponding drugs, and those in the normal control group and model group were given an equal volume of distilled water. After 14 consecutive days of prophylactic gavage, the rat model of common carotid artery thrombosis was established with 45% FeCl3 solution, and the blood vessels were collected and the wet weight of thrombus was weighed by an electronic balance (precision of 1/10 000). The thrombosis in the common carotid artery of each group of rats was observed by hematoxylin-eosin staining. The plasma levels of von Willebrand factor (vWF), platelet endothelial cell adhesion molecule-1 (PECAM-1), tissue-type plasminogen activator (t-PA), and plasminogen activator inhibitor-1 (PAI-1) were determined by enzyme-linked immunosorbent assay. An endothelial cell injury model was established by treating EA.hy926 human umbilical vein endothelial cells with TNF-α. The cell counting kit-8 method was used to screen the intervention concentrations of QXJYG. Western blot was employed to determine the protein levels of TF, TFPI, Akt, p-Akt, NF-κB p65, and p-NF-κB p65 in each group of cells. ResultsThe animal experiment showed that compared with the normal control group, the model group showed an increase in carotid artery thrombus weight (P<0.05), with unclear vascular structure and extensive thrombosis in the lumen. In addition, the plasma levels of vWF, PECAM-1, and PAI-1 were elevated, while the t-PA level became lowered (P<0.05) in the model group. Compared with the model group, the aspirin and QXJYG groups showed reductions in the weight of FeCl3-induced carotid artery thrombi (P<0.05) and thrombosis in the lumen, declines in plasma levels of PECAM-1 and PAI-1, and an elevation in the t-PA level (P<0.05). Moreover, the QXJYG groups showed reductions in the plasma level of vWF (P<0.05), which, however, had no significant difference between the aspirin group and the model group. The cell experiments indicated that 31.25, 62.5, 125, 250, 500 mg·L-1 QXJYG had no effect on the viability of EA.hy926 cells. Therefore, 250, 500 mg·L-1 QXJYG were selected as the intervention concentrations for subsequent experiments. Western blotting results showed that compared with the control group, the TNF-α stimulation downregulated the expression of TFPI (P<0.05), upregulated the expression of TF, and increased the ratios of p-Akt/Akt and p-NF-κB p65/NF-κB p65 (P<0.05) in EA.hy926 cells. Compared with the model group, the intervention with QXJYG upregulated the expression of TFPI (P<0.05), inhibited the expression of TF, and decreased the ratios of p-Akt/Akt and p-NF-κB p65/NF-κB p65 (P<0.05). ConclusionQXJYG has the effect of inhibiting thrombosis and regulating the expression of TF and TFPI in endothelial cells exposed to TNF-α by suppressing the abnormal activation of the Akt/NF-κB signaling pathway.
3.Syndrome and Pathogenesis Treated by Mahuang Shengmatang
Xinyu WAN ; Yin CHENG ; Wenbo GAO ; Zhiguo ZHANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(3):253-258
The prescription Mahuang Shengmatang in the Treatise on Cold Damage (《伤寒论》) has sparked considerable debate among medical practitioners throughout history, with varying opinions on its indications, pathogenesis described in the text, principle of compatibility, and clinical applications. Both ancient and modern interpreters of Mahuang Shengmatang often focus on herbal compatibility as a primary approach to deduce the pathogenesis and treatment methods. Building upon this foundation, this paper utilizes herbal dosage as a clue to discern the primary and secondary herbs in the prescription. It further analyzes the principle of compatibility based on herbal indications, summarizing the indications and therapeutic principles of this prescription. Ultimately, it reveals the underlying pathogenesis reflected in the text. The internal closure of heat and toxin leads to the stagnation of Qi, preventing Yang Qi from reaching the extremities and causing cold hands and feet. When the pathogenic Qi finds no outlet, it floods both the upper and lower regions of the body, attacking the throat and causing cough with expectoration of pus and blood, and descending to the large intestine to consume Yin fluids, resulting in persistent diarrhea. Based on this pathogenesis, the paper expands the scope of symptoms and signs associated with the prescription, providing a more detailed portrayal of the applicable patient population and enhancing the basis for clinical prescription references. Additionally, the paper presents considerations on several controversial topics, suggesting that the "lower pulse" described in the text refers to the lower limb arterial pulsation, and the symptoms and signs resemble those of septic shock in modern medicine. Therefore, Mahuang Shengmatang should be categorized as a prescription for treating warm diseases and it is not developed by ZHANG Zhongjing. By employing a detailed discussion on the syndrome, pathogenesis, and clinical application in the texts of Mahuang Shengmatang from the dosage, principle of compatibility, and herbal indications, this paper not only enriches the theoretical foundation of Mahuang Shengmatang but also provides a comprehensive perspective and fresh ideas for understanding its clinical application.
4.Exploring the Application of "Cleaning Spleen and Restoring Defensive Qi" Method in Treatment of Pancreatic Cancer based on Neutrophil Extracellular Traps Abnormal Accumulation
Chuanlong ZHANG ; Mengqi GAO ; Yi LI ; Xiaochen JIANG ; Songting SHOU ; Bo PANG ; Baojin HUA
Journal of Traditional Chinese Medicine 2025;66(1):30-33
The abnormal accumulation of neutrophil extracellular traps (NETs) can promote the initiation and progression of pancreatic cancer, which is considered a potential therapeutic target for this disease. The Miraculous Pivot·Inquiry About Statement (《灵枢·口问》) have recorded the concept of "defensive qi stagnation". Based on the recognition that the function of defensive qi is similar to the immune function of neutrophils, and combining traditional Chinese medicine theory with clinical practice, it is proposed that the abnormal accumulation of NETs may be a pathological product of "defensive qi stagnation", with the spleen being the critical site of pathology. Further exploring the application strategy of cleaning spleen and restoring defensive qi method in pancreatic cancer treatment, it is proposed to employ three approaches such as dredging method to eliminate spleen stagnation and inhibit pancreatic cancer proliferation, cleaning method to remove spleen dampness and suppress the inflammatory micro-environment, and tonifying method to strengthen Weiqi and to improve the immune microenvironment, which aims to provide new insights for the clinical treatment of pancreatic cancer with traditional Chinese medicine.
5.Nuangong Tiaojing Decoction (暖宫调经汤) Combined with Ginger Moxibustion at Baliao (八髎) Acupoints for Patients with Premature Ovarian Failure of Yang-Deficiency Uterus-Coldeness Syndrome:65 Cases Clinical Observation
Yuqiong MENG ; Qiang GAO ; Tingting ZHAI ; Zhihong YANG
Journal of Traditional Chinese Medicine 2025;66(1):59-64
ObjectiveTo explore the clinical effectivess of Nuangong Tiaojing Decoction (暖宫调经汤) combined with ginger moxibustion at Baliao acupoints (Eight Bone-Holes) for treating premature ovarian failure (POF) of yang-deficiency uterus-coldeness syndrome, based on conventional western medical treatment. MethodsA total of 130 patients diagnosed with POF of yang-deficiency uterus-coldeness syndrome were divided into a treatment group and a control group, with 65 cases in each group, based on patient preference. The control group received conventional western medicine treatment, while the treatment group was additionally given Nuangong Tiaojing Decoction orally one dose per day and underwent ginger moxibustion at Baliao points once daily. Both groups were treated for three months. Before and after treatment, ovarian function indicators, including ovarian volume, follicle count, ovarian peak systolic velocity (PSV), and endometrial thickness were assessed, and serum sex hormone levels including progesterone (P), estradiol (E2), follicle-stimulating hormone (FSH), luteinizing hormone (LH), prolactin (PRL), and testosterone (T), as well as anti-Müllerian hormone (AMH) levels were also measured. Additionally, traditional Chinese medicine (TCM) syndrome scores were evaluated, covering symptoms such as reduced libido, lumbosacral pain, fear of cold with cold extremities, fatigue, dull complexion, lower abdominal distension and pain, pale tongue, and deep and rough pulse. After treatment, clinical effectiveness and safety were determined. ResultsCompared with the pre-treatment levels within group, both groups showed significant increases in ovarian volume, follicle count, PSV, endometrial thickness, and levels of P, E2, and AMH after treatment, while the levels of FSH, LH, PRL, and T, as well as scores of lumbosacral pain, reduced libido, dull complexion, deep and rough pulse, and total TCM syndrome scores significantly reduced after treatment. Furthermore, the treatment group exhibited higher ovarian volume, follicle count, PSV, endometrial thickness, and levels of P, E2, and AMH compared to the control group, while FSH, LH, PRL, T levels, and all symptom scores, as well as total TCM syndrome scores, were significantly lower in the treatment group than in the control group (P<0.05 or P<0.01). The clinical effectiveness in treatment group was 96.92% (63/65), significantly higher than 81.54% (53/65) in control group (P<0.05). The incidence of adverse reactions in the treatment group was 26.15% (17/65), while in the control group it was 32.31% (21/65), with no statistically significant difference (P>0.05). ConclusionOn the basis of conventional western medicine treament, Nuangong Tiaojing Decoction combined with ginger moxibustion at Baliao acupoints for patients with POF of yang-deficiency uterus-coldeness syndrome can significantly improve ovarian function and clinical symptoms, regulate hormone levels, and thereby enhance clinical effectiveness.
6.Effects of Shenfuhuang Formula (参附黄配方) on Potential Targets of Action in the Brain Tissue of Sepsis Model Mice:Transcriptomics-Based Exploration
Yuchen WANG ; Xuerui WANG ; Xiaolong XU ; Jingxia ZHAO ; Jiabo WANG ; Yuan GAO ; Weijun KONG ; Qingquan LIU
Journal of Traditional Chinese Medicine 2025;66(1):65-70
ObjectiveTo investigate the possible mechanism of Shenfuhuang Formula (参附黄配方) in prevention and treatment of epsis-associated encephalopathy from the perspective of brain genomics. MethodsC57BL/6 mice were randomly divided into sham surgery group, sepsis group, and Shenfuhuang group, with 20 mice in each group. The sepsis group and Shenfuhuang group were induced to develop sepsis by cecal ligation and puncture (CLP) procedure. At 4 hours after modelling, Shenfuhuang group were gavaged with 2.5 g/(kg·d) of Shenfuhuang Formula, 0.5 ml each time, at 12 hours intervals, for a total of 4 times after modelling. Sepsis group and sham surgery group were given 0.5 ml of purified water orally. At 48 hours after modeling, the transcriptome sequencing was used to explore the differential gene expression in the effects of Shenfuhuang Formula on the brain regions of septic mice, and real-time PCR and ELISA were later used to further validate the differential gene and proteins expression. ResultsA total of 4605 genes were differentially expressed in Shenfuhuang group compared with sepsis group, of which 2353 genes were up-regulated and 2252 genes were down-regulated. According to the results of previous publications, six key genes were screened, including serine/threonine-protein kinase (Nek1), myelin-associated glycoprotein (Mag), endothelial cell-specific tyrosine kinase receptor (Tek), a disintegrin and metalloproteinase with thrombospondin motifs 20 (Adamts20), lymphocyte antigen 86 (Ly86), and E3 ubiquitin-protein ligase (Traip). Further genetic and protein validation revealed that, compared to the sham surgery group, the mRNA levels and corresponding protein levels of Nek1, Mag, Tek, Adamts20, Ly86, and Traip in the brain tissue of septic mice significantly reduced (P<0.05). In comparison to the sepsis group, Shenfuhuang group showed significantly increased mRNA levels and corresponding protein levels of Nek1, Mag, Tek, Adamts20, Ly86, and Traip (P<0.05). ConclusionThe potential therapeutic targets of Shenfuhuang Formula for treating sepsis-associated encephalopathy may be related to the Nek1, Mag, Tek, Adamts20, Ly86, and Traip genes and their encoded proteins.
7.Clinical Advantages and Key Research Points of Traditional Chinese Medicine in the Treatment of Atrial Fibrillation
Cong SUN ; Yujiang DONG ; Hongmei GAO ; Qing WEI ; Menghe ZHANG ; Xiaojing SHI ; Liya FENG
Journal of Traditional Chinese Medicine 2025;66(2):133-138
Traditional Chinese medicine (TCM) therapy has unique clinical advantages in the treatment of atrial fibrillation, mainly reflected in five aspects, improving quality of life, enabling early diagnosis and treatment, promoting cardiac rehabilitation, making up for the limitations of Western medicine, and improving the success rate of catheter ablation. However, there is insufficient evidence in current clinical research. Based on the current status of TCM research in the treatment of atrial fibrillation, it is suggested that future studies should focus on standardized research on syndrome differentiation and classification. This can be achieved through clinical epidemiological surveys, expert consensus, and other methods to establish a unified syndrome differentiation and classification standard for atrial fibrillation. Clinical efficacy evaluation indicators should be standardized, and core outcome measures for clinical research on TCM treatment of atrial fibrillation should be developed through systematic reviews, patient interviews, and other methods. Additionally, clinical research design, implementation, and data management should be improved. By leveraging modern information technologies such as artificial intelligence, the scientific and standardized nature of TCM intervention research on atrial fibrillation can be enhanced, ultimately improving the quality of research.
8.Ideas of Traditional Chinese Medicine Treatment of Pancreatic Endocrine and Exocrine Co-Morbidities from the Attributes of Zang-Fu Organs of Pancreas
Yulin LENG ; Jiacheng YIN ; Xianglong LI ; Jiahong ZHANG ; Yi SU ; Hong GAO ; Chunguang XIE ; Xiaoxu FU
Journal of Traditional Chinese Medicine 2025;66(2):145-149
Based on advancements in modern medical research regarding the intricate connection between the endocrine and exocrine functions of the pancreas, as well as the relationship between pancreatic functions and traditional Chinese medicine (TCM) spleen system, this paper discussed the categorization of the pancreas. It is proposed that the pancreas is neither a true zang organ nor a fu organ, but possessed the attributes of an extraordinary fu-organ and can be classified under the spleen. The spleen governs transportation and transformation, ascent of the clear and dispersion of essence, which encompasses the endocrine and exocrine functions, and pancreatic enzymes and glucose-regulating hormones form the material basis for the spleen's function of dispersing essence. Diseases of the pancreas exhibit characteristics of both zang-organ deficiency and fu-organ excess, so treatment should simultaneously supplement zang-organ disease and regulate fu-organ disease when pancreas showing endocrine and exocrine co-morbidities, with focus on restoring the pancreas (spleen)'s dispersing essence function. Therapeutic strategies include supplementing spleen qi, nourishing spleen yin to strengthen spleen earth, unblocking spleen collaterals, raising spleen yang, and removing spleen turbidity to support the spleen's dispersing essence function, so as to replenish the essential qi of zang-fu organs, ensure their distribution throughout the body, and improve the endocrine and exocrine functions of the pancreas.
9.Prevalence and influencing factors of school bullying experienced by primary and middle school students
ZHU Yunjiao ; GU Fang ; MENG Jia ; LI Juanjuan ; SHEN Yu ; GAO Lei
Journal of Preventive Medicine 2025;37(1):1-6
Objective:
To investigate the situation and influencing factors of school bullying experienced by primary and middle school students, so as to provide the basis for formulating school bullying intervention measures and promoting students' physical and mental health development.
Methods:
All the counties (cities, districts) in Zhejiang Province were stratified to urban and suburban areas, primary, junior high and senior high school students were selected using a stratified cluster sampling method. Basic information, lifestyle and school bullying were collected through questionnaire surveys. Factors affecting school bullying experienced by primary and middle school students were analyzed using a multivariable logistic regression model.
Results:
Totally 137 846 valid questionnaires were recovered, with an effective recovery rate of 97.17%. There were 72 526 males (52.61%) and 65 320 females (47.39%). There were 47 561 primary school students (34.50%), 47 701 junior high school students (34.61%) and 42 584 senior high school students (30.89%). A total of 3 987 students suffered from school bullying, accounting for 2.89%. The proportions of being maliciously teased, being intentionally excluded from group activities/isolated, being teased about physical defects or appearance, being hit/kicked/pushed/shoved/locked in a room, being threatened, and being extorted for money were 2.04%, 1.18%, 1.11%, 0.86%, 0.84% and 0.83%, respectively. Multivariable logistic regression analysis showed that the students who were males (OR=1.122, 95%CI: 1.048-1.202), lived in suburban areas (OR=1.322, 95%CI: 1.233-1.418), lived in areas with medium (OR=1.086, 95%CI: 1.006-1.173) or underdeveloped (OR=1.298, 95%CI: 1.191-1.415) economic level, had higher academic levels (junior high school, OR=1.380, 95%CI: 1.270-1.499; senior high school, OR=1.210, 95%CI: 1.083-1.351), lived on campus (OR=1.489, 95%CI: 1.372-1.616), engaged in fights (OR=6.029, 95%CI: 5.585-6.509), attempted to smoke (OR=1.320, 95%CI: 1.128-1.545), drank (OR=1.735, 95%CI: 1.575-1.912), were scolded and beaten by parents (OR=1.972, 95%CI: 1.822-2.135) and were obese (OR=1.240, 95%CI: 1.132-1.360) were more likely to experience school bullying.
Conclusion
The harm of school bullying to the physical and mental health of primary and middle school students should be taken seriously, and active policy measures should be adopted to strengthen intervention.
10.Availability and use of child safety seats among children aged 0-3 years
CHEN Bo ; WANG Xihui ; QIU Fengqian ; YU Yan ; GAO Shuna ; HE Lihua ; LI Weiyi ; JI Yunfang ; CHEN Weihua
Journal of Preventive Medicine 2025;37(1):21-25
Objective:
To investigate the availability and use of child safety seats among children aged 0-3 years, so as to provide the basis for improving riding safety for children.
Methods:
Parents of children aged 0-3 years in Huangpu District, Shanghai Municipality, were recruited using the stratified multistage random sampling method from May to July 2024. Demographic information, family travel patterns, the use of child safety seat and related health beliefs were collected using questionnaire surveys. Factors affecting the use of child safety seats were identified using a multivariable logistic regression model.
Results:
Totally 514 valid questionnaires were recovered, with an effective rate of 96.98%. The respondents included 122 fathers (23.74%) and 392 mothers (76.26%), with a median age of 34.00 (interquartile range, 5.00) years. There were 446 families equipping with child safety seats, accounting for 86.77%; and 169 families using child safety seats, accounting for 32.88%. Multivariable logistic regression analysis showed that the parents who had children aged >1-2 years (OR=0.597, 95%CI: 0.366-0.973), travelled 2-4 times per month (OR=0.359, 95%CI: 0.213-0.607) or once per month or less (OR=0.384, 95%CI: 0.202-0.729), and scored high in perceived barrier (OR=0.634, 95%CI: 0.486-0.827) were less likely to use child safety seats; the parents who had children with local household registration (OR=2.506, 95%CI: 1.356-4.633), travelled 5-<10 km (OR=1.887, 95%CI: 1.148-3.101) or ≥10 km (OR=2.319, 95%CI: 1.355-3.967), always wore seat belts (OR=2.342, 95%CI: 1.212-4.524), scored high in perceived susceptibility (OR=1.392, 95%CI: 1.091-1.778) and self-efficacy (OR=1.413, 95%CI: 1.156-1.727) were more likely to use child safety seats.
Conclusions
Equipping family cars with child safety seats and using them can prevent and reduce traffic injuries among children aged 0-3 years. It is recommended to strengthen publicity to promote the use of child safety seats.


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