1.Liuwei Dihuangwan Promote Mitophagy to Modulate Neuroinflammation and Behavioral Impairments in Rat Model of Autism Spectrum Disorder (ASD)
Pengjue HUANG ; Mingyue JIANG ; Ji WU ; Niya YIN ; Lei OUYANG ; Qinquan ZHU ; Di ZHANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(2):52-60
ObjectiveTo observe the effect of Liuwei Dihuangwan on behavioral impairments in the rat model of autism spectrum disorder (ASD) and explore the mechanism of action. MethodsTwelve SD pregnant rats were intraperitoneally injected with valproic acid (VPA) (10 rats) or normal saline (2 rats), and male offspring were selected to establish the model of ASD and the control rats. Rats were randomly assigned into model, low-dose (0.75 g·kg-1) and high-dose (1.5 g·kg-1) Liuwei Dihuangwan, vitamin D (positive drug, 3.7×10-5 g·kg-1), and blank groups. Each group was administrated with the corresponding concentration of drugs or the same volume of normal saline by gavage for 2 weeks. After the intervention, the three-chamber social test was conducted to evaluate social interaction and social preference. The open field test was carried out to observe spontaneous behavior and anxiety state. Hematoxylin-eosin staining (HE) was used to observe the pathological changes of the prefrontal tissue. Transmission electron microscopy was employed to observe the ultrastructure of mitochondria in prefrontal neurons. Immunofluorescence was used to detect the expression of ionized calcium-binding adapter molecule-1 (Iba-1) in the prefrontal tissue. Enzyme-linked immunosorbent assay (ELISA) was adopted to measure the levels of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6). Western blot was employed to assess the expression differences of phosphorylated adenosine monophosphate-activated protein kinase (p-AMPK), adenosine monophosphate-activated protein kinase (AMPK), phosphorylated Unc-51-like autophagy-activating kinase 1 (p-ULK1), Unc-51-like autophagy-activating kinase 1 (ULK1), and FUN14 domain-containing protein 1 (FUNDC1). ResultsCompared with the blank group, the model group spent less time sniffing stranger 1 and stranger 2 in the three-chamber social test (P<0.01) and showed reductions in the total distance traveled, average speed, distance traveled in the central area, and time spent in the central area in the open field test (P<0.01). In addition, the model group showed extensive apoptosis of neurons, with shrunken nuclei and red-stained cytoplasm, and extensive necrosis of neurons in the prefrontal tissue, mitochondrial swelling, decreased matrix density, disrupted cristae, and autophagic lysosomes in neurons, increases in the rate of Iba-1 positive cells in the prefrontal area (P<0.01) and the levels of TNF-α and IL-6 (P<0.01), and down-regulation in the expression of p-AMPK/AMPK, p-ULK1/ULK1, and FUNDC1 (P<0.01). Compared with the model group, low-dose and high-dose Liuwei Dihuangwan and the vitamin D prolonged the time spent sniffing stranger 1 and stranger 2 in the three-chamber social test (P<0.05, P<0.01), increased the total distance traveled, average speed, distance traveled in the central area, and time spent in the central area in the open field test (P<0.05, P<0.01), restored the morphology of neurons in the prefrontal tissue, decreased the number of apoptotic cells, alleviated the swelling of mitochondria in neurons, increased the matrix density, mitigated the fragmentation and disorder of cristae, and increased the number of autophagosomes. Moreover, the drugs decreased the rate of Iba-1 positive cells in the prefrontal area (P<0.01), lowered the levels of TNF-α and IL-6 (P<0.01), and up-regulated the expression of p-AMPK/AMPK, p-ULK1/ULK1, and FUNDC1 (P<0.01). ConclusionLiuwei Dihuangwan ameliorate autism-like behaviors and reduce neuronal apoptosis and neuroinflammatory damage in the rat model of ASD by promoting mitophagy mediated by the AMPK/ULK1/FUNDC1 pathway.
2.Neuroprotective Mechanism of Yifei Xuanfei Jiangzhuo Prescription on VaD Rats Based on NF-κB/NLRP3 Signaling Pathway
Bingmao YUAN ; Wei CHEN ; Xiu LAN ; Lingfei JIANG ; Lin WU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(3):88-96
ObjectiveTo investigate the molecular mechanism by which Yifei Xuanfei Jiangzhuo prescription regulates the nuclear factor-κB (NF-κB)/NOD-like receptor protein 3 (NLRP3) signaling pathway to improve neuronal function in vascular dementia (VaD) rats. MethodsA VaD model was established by intermittently clamping the bilateral common carotid arteries (CCA) combined with bilateral vascular occlusion (2-VO). Eighty-four SD rats were randomly divided into a blank group, sham group, model group, piracetam group (0.2 g·kg-1), and low-, medium-, and high-dose Yifei Xuanfei Jiangzhuo prescription groups (6.09, 12.18, and 24.36 g·kg-1). Drug administration began on day 7 after surgery, once daily for 28 consecutive days. Behavioral experiments were used to evaluate learning and spatial memory. Hematoxylin-eosin (HE) staining was applied to observe pathological morphological changes in the CA1 region of the hippocampus. Transmission electron microscopy was used to examine the ultrastructure of hippocampal neurons. Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) was used to detect neuronal apoptosis in the CA1 region. Immunohistochemistry was performed to determine the positive expression rate of neuronal nuclear antigen (NeuN). Immunofluorescence single staining was used to assess nuclear expression of NF-κB p65 in brain tissue. Western blot was used to detect the protein expression levels of inhibitor of κB kinase (IKK), NF-κB p65, NLRP3, Caspase-1, apoptosis-associated speck-like protein (ASC), and interleukin-1β (IL-1β). ResultsCompared with the blank group, the model group showed a significant reduction in platform-crossing frequency (P0.01), aggravated hippocampal injury, a significant increase in neuronal apoptosis (P0.05), decreased NeuN positivity in the CA1 region (P0.05), increased nuclear expression of NF-κB p65 (P0.05), and significantly elevated expression of p-IKK, p-NF-κB p65, NLRP3, cleaved Caspase-1, ASC, and cleaved IL-1β (P0.05). Compared with the model group, all drug-treated groups improved learning and spatial memory in VaD rats, alleviated hippocampal pathological injury and neuronal apoptosis, and protected neuronal ultrastructure. Yifei Xuanfei Jiangzhuo prescription at doses of 12.18 and 24.36 g·kg-1 reduced hippocampal expression levels of p-IKK, p-NF-κB p65, NLRP3, Caspase-1, ASC, and cleaved IL-1β in VaD rats (P0.05), showing dose-dependent inhibition of the NF-κB/NLRP3 signaling pathway. ConclusionYifei Xuanfei Jiangzhuo prescription may exert neuroprotective effects by regulating the NF-κB/NLRP3 signaling pathway, thereby reducing neuroinflammation and inhibiting hippocampal neuronal apoptosis.
3.Yimei Baijiang Formula Treats Colitis-associated Colorectal Cancer in Mice via NF-κB Signaling Pathway
Qian WU ; Xin ZOU ; Chaoli JIANG ; Long ZHAO ; Hui CHEN ; Li LI ; Zhi LI ; Jianqin LIU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(3):119-130
ObjectiveTo explore the effects of Yimei Baijiang formula (YMBJF) on colitis-associated colorectal cancer (CAC) and the nuclear factor kappaB (NF-κB) signaling pathway in mice. MethodsSixty male Balb/c mice of 4-6 weeks old were randomized into 6 groups: Normal, model, capecitabine (0.83 g
4.Neuroprotective Mechanism of Yifei Xuanfei Jiangzhuo Prescription on VaD Rats Based on NF-κB/NLRP3 Signaling Pathway
Bingmao YUAN ; Wei CHEN ; Xiu LAN ; Lingfei JIANG ; Lin WU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(3):88-96
ObjectiveTo investigate the molecular mechanism by which Yifei Xuanfei Jiangzhuo prescription regulates the nuclear factor-κB (NF-κB)/NOD-like receptor protein 3 (NLRP3) signaling pathway to improve neuronal function in vascular dementia (VaD) rats. MethodsA VaD model was established by intermittently clamping the bilateral common carotid arteries (CCA) combined with bilateral vascular occlusion (2-VO). Eighty-four SD rats were randomly divided into a blank group, sham group, model group, piracetam group (0.2 g·kg-1), and low-, medium-, and high-dose Yifei Xuanfei Jiangzhuo prescription groups (6.09, 12.18, and 24.36 g·kg-1). Drug administration began on day 7 after surgery, once daily for 28 consecutive days. Behavioral experiments were used to evaluate learning and spatial memory. Hematoxylin-eosin (HE) staining was applied to observe pathological morphological changes in the CA1 region of the hippocampus. Transmission electron microscopy was used to examine the ultrastructure of hippocampal neurons. Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) was used to detect neuronal apoptosis in the CA1 region. Immunohistochemistry was performed to determine the positive expression rate of neuronal nuclear antigen (NeuN). Immunofluorescence single staining was used to assess nuclear expression of NF-κB p65 in brain tissue. Western blot was used to detect the protein expression levels of inhibitor of κB kinase (IKK), NF-κB p65, NLRP3, Caspase-1, apoptosis-associated speck-like protein (ASC), and interleukin-1β (IL-1β). ResultsCompared with the blank group, the model group showed a significant reduction in platform-crossing frequency (P0.01), aggravated hippocampal injury, a significant increase in neuronal apoptosis (P0.05), decreased NeuN positivity in the CA1 region (P0.05), increased nuclear expression of NF-κB p65 (P0.05), and significantly elevated expression of p-IKK, p-NF-κB p65, NLRP3, cleaved Caspase-1, ASC, and cleaved IL-1β (P0.05). Compared with the model group, all drug-treated groups improved learning and spatial memory in VaD rats, alleviated hippocampal pathological injury and neuronal apoptosis, and protected neuronal ultrastructure. Yifei Xuanfei Jiangzhuo prescription at doses of 12.18 and 24.36 g·kg-1 reduced hippocampal expression levels of p-IKK, p-NF-κB p65, NLRP3, Caspase-1, ASC, and cleaved IL-1β in VaD rats (P0.05), showing dose-dependent inhibition of the NF-κB/NLRP3 signaling pathway. ConclusionYifei Xuanfei Jiangzhuo prescription may exert neuroprotective effects by regulating the NF-κB/NLRP3 signaling pathway, thereby reducing neuroinflammation and inhibiting hippocampal neuronal apoptosis.
5.Yimei Baijiang Formula Treats Colitis-associated Colorectal Cancer in Mice via NF-κB Signaling Pathway
Qian WU ; Xin ZOU ; Chaoli JIANG ; Long ZHAO ; Hui CHEN ; Li LI ; Zhi LI ; Jianqin LIU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(3):119-130
ObjectiveTo explore the effects of Yimei Baijiang formula (YMBJF) on colitis-associated colorectal cancer (CAC) and the nuclear factor kappaB (NF-κB) signaling pathway in mice. MethodsSixty male Balb/c mice of 4-6 weeks old were randomized into 6 groups: Normal, model, capecitabine (0.83 g
6.Epidemiological analysis of a cluster outbreak of pulmonary tuberculosis among grade 12 students from a boaring high school in Chongqing
LEI Rongrong, FENG Xinyu, XIA Siyue, JIANG Chuan, ZHANG Ting, WU Chengguo
Chinese Journal of School Health 2026;47(1):113-116
Objective:
To analyze the process of handling a pulmonary tuberculosis(TB) outbreak among senior high school students in a boarding school in Chongqing, as well as to investigate the underlying causes of the outbreak, so as to provide evidence to inform TB prevention and control strategies in school settings.
Methods:
From November 2023 to April 2024, an epidemiological investigation was conducted into the TB outbreak in a grade 12 class from a boarding high school. Suspected cases were screened using symptom screening, tuberculin skin test (TST), and chest X-ray examinations. Confirmed cases underwent individual epidemiological interviews and sputum culture; Cultured positive mycobacterial strains were subjected to whole genome sequencing after identification as Mycobacterium tuberculosis.
Results:
A total of 10 active pulmonary TB cases were identified, all from the same class, yielding a student attack rate of 16.67%. Three isolates were culture positive, as well as all strains were of L2 type,and the WGS analysis of the strains suggested a common transmission chain. Excluding the index case, four additional cases were detected through symptom driven health care visits. Notably, 70% of patients presented with "chest tightness and chest pain" symptoms, and 50% had "cough" symptoms,but none were detected during morning health checks or tracking of absences due to illness. A total of 326 contacts were identified and underwent three rounds of screening and one follow up examination. In the initial screening, 35 close contacts from the same class showed strong TST positivity, corresponding to a strong positivity rate of 55.56%, significantly higher than the 20.76% observed among casual contacts ( χ 2=29.80, P <0.01). Among the 35 strongly TST positivvity close contacts and five individuals with moderate TST positivity whose induration had increased by ≥10 mm over two years, none received timely preventive treatment initially; five of them were subsequently diagnosed with active TB within three months. Following this, 25 individuals initiated preventive therapy, resulting in a preventive treatment initiation rate of 62.50%. Among TST negative classmates who converted to strong positivity on repeat TST testing at three months, 75.00% started preventive treatment, but only 22.22% completed the full course.
Conclusion
Inadequate implementation of morning health checks and cause tracking for absenteeism due to illness, poorly standardized screening procedures, and delayed preventive treatment may have been key factors contributing to the spread of the outbreak.
7.Potential Mechanism of Zuojinwan in Improving Liver Fibrosis Based on Hepatic Tissue Metabolomics
Yiting JIANG ; Kexin LIU ; Yixi QIAN ; Rui ZHANG ; Feng ZHANG ; Hongyan WU ; Li CHEN
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(6):54-61
ObjectiveThis study aims to elucidate the potential mechanism of Zuojinwan in improving liver fibrosis through hepatic tissue metabolomics analysis. MethodsTwenty-four mice were randomly allocated into normal group, model group , positive drug group (silymarin, 100 mg·kg-1), and Zuojinwan group (Zuojinwan solution, 2.5 g·kg-1), with per group six mice. Liver fibrosis model was induced via intraperitoneal injection of olive oil solution with 10% carbon tetrachloride (CCl4) (0.5 μL·g-1, three times weekly for 8 weeks) in all groups except the normal group. During the final 4 weeks, the silymarin group received silymarin (100 mg·kg-1) by gavage thrice weekly, while the Zuojinwan group was administered Zuojinwan solution (2.5 g·kg-1) under the same regimen. After the last administration, the levels of liver fibrosis indicators and liver injury markers in serum were detected. The pathological morphological changes of the liver tissues were observed. The levels of liver fibrosis markers α-smooth muscle actin (α-SMA) and Collagen Ⅰ(ColⅠ) were detected. Metabolomics was analyzed on mice's liver tissues. The mice's serum was collected for metabolomics analysis. ResultsCompared with the model group, Zuojinwan significantly improved indicators related to liver fibrosis and liver injury. Compared with the normal group, the model group showed significantly elevated levels of fibrosis markers such as laminin (LN), hyaluronic acid (HA), procollagen typeⅢ (PC-Ⅲ), and type Ⅳ Collagen (Ⅳ-C), while liver injury indicators such as alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), lactate dehydrogenase (LDH), and total bilirubin (TBIL), exhibited a marked upward trend (P<0.05). Compared with the model group, the silymarin group showed a significant decrease in the aforementioned indicators (P<0.05). Notably, compared with the model group, the Zuojinwan group exhibited a significant reduction in all these indicators (P<0.05), with efficacy comparable to that of the silymarin group. Zuojinwan reduced mRNA and protein levels of α-SMA and ColⅠ in the liver tissue. Metabolomics results revealed that compared with the model group, Zuojiinwan significantly reduced levels of glucose metabolism-related metabolites such as D-fructose 1,6-bisphosphate (FBP), nicotinamide adenine dinucleotide phosphate (NADPH), sodium beta-D-fructose 6-phosphate (F6P), dihydroxyacetone phosphate (DHAP), fumaric acid, and D-glucose 6-phosphate (G6P) (P<0.05). Serum enzyme-linked immunosorbent assay (ELISA) was used to detect glucose metabolism indicators and further validate the regulatory effect of Zuojinwan on glucose metabolism. ConclusionThese results suggest that Zuojinwan may improve liver fibrosis by regulating the dysregulated levels of glucose metabolism during the progression of liver fibrosis.
8.Revealing VCAN as a Potential Common Diagnostic Biomarker of Renal Tubules and Glomerulus in Diabetic Kidney Disease Based on Machine Learning, Single-Cell Transcriptome Analysis and Mendelian Randomization
Li JIANG ; Jie JIAN ; Xulin SAI ; Xiai WU
Diabetes & Metabolism Journal 2025;49(3):407-420
Background:
Diabetic kidney disease (DKD) is recognized as a significant complication of diabetes mellitus and categorized into glomerular DKDs and tubular DKDs, each governed by distinct pathological mechanisms and biomarkers.
Methods:
Through the identification of common features observed in glomerular and tubular lesions in DKD, numerous differentially expressed gene were identified by the machine learning, single-cell transcriptome and mendelian randomization.
Results:
The diagnostic markers versican (VCAN) was identified, offering supplementary options for clinical diagnosis. VCAN significantly highly expressed in glomerular parietal epithelial cell and proximal convoluted tubular cell. It was mainly involved in the up-regulation of immune genes and infiltration of immune cells like mast cell. Mendelian randomization analysis confirmed that serum VCAN protein levels were a risky factor for DKD, while there was no reverse association. It exhibited the good diagnostic potential for estimated glomerular filtration rate and proteinuria in DKD.
Conclusion
VCAN showed the prospects into DKD pathology and clinical indicator.
9.Rapid Video Analysis for Contraction Synchrony of Human Induced Pluripotent Stem Cells-Derived Cardiac Tissues
Yuqing JIANG ; Mingcheng XUE ; Lu OU ; Huiquan WU ; Jianhui YANG ; Wangzihan ZHANG ; Zhuomin ZHOU ; Qiang GAO ; Bin LIN ; Weiwei KONG ; Songyue CHEN ; Daoheng SUN
Tissue Engineering and Regenerative Medicine 2025;22(2):211-224
BACKGROUND:
The contraction behaviors of cardiomyocytes (CMs), especially contraction synchrony, are crucial factors reflecting their maturity and response to drugs. A wider field of view helps to observe more pronounced synchrony differences, but the accompanied greater computational load, requiring more computing power or longer computational time.
METHODS:
We proposed a method that directly correlates variations in optical field brightness with cardiac tissue contraction status (CVB method), based on principles from physics and photometry, for rapid video analysis in wide field of view to obtain contraction parameters, such as period and contraction propagation direction and speed.
RESULTS:
Through video analysis of human induced pluripotent stem cell (hiPSC)-derived CMs labeled with green fluorescent protein (GFP) cultured on aligned and random nanofiber scaffolds, the CVB method was demonstrated to obtain contraction parameters and quantify the direction and speed of contraction within regions of interest (ROIs) in wide field of view. The CVB method required less computation time compared to one of the contour tracking methods, the LucasKanade (LK) optical flow method, and provided better stability and accuracy in the results.
CONCLUSION
This method has a smaller computational load, is less affected by motion blur and out-of-focus conditions, and provides a potential tool for accurate and rapid analysis of cardiac tissue contraction synchrony in wide field of view without the need for more powerful hardware.
10.Platelet-rich plasma injection combined with warm acupuncture and moxibustion for treating patients with knee osteoarthritis and cold dampness obstruction syndrome
Xiang SHANG ; Fei WANG ; Qiqi YANG ; Tianxin JIANG ; Fen ZHANG ; Sanbing WU ; Yonghui YANG ; Fei LI
Journal of Beijing University of Traditional Chinese Medicine 2025;48(2):270-279
Objective:
To determine the clinical efficacy of platelet-rich plasma (PRP) injection combined with warm acupuncture and moxibustion for treating patients with knee osteoarthritis and cold dampness obstruction syndrome.
Methods:
One hundred and twenty-eight patients with knee osteoarthritis and cold dampness obstruction syndrome who visited the Rehabilitation Department and Orthopedics Department of the Second Affiliated Hospital of Anhui University of Chinese Medicine from January 2023 to March 2024 and who met the inclusion and exclusion criteria were randomly divided into an experimental (n=64) and control group (n=64) using the random number table method. The experimental group was treated with PRP injection combined with warm acupuncture and moxibustion, whereas the control group was treated with normal saline injection combined with warm acupuncture and moxibustion treatment. PRP and normal saline injections were administered once every two weeks, a total of four times. Patients were treated with warm acupuncture and moxibustion once a day, six times a week, for four consecutive weeks. The Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC), Traditional Chinese Medicine (TCM) syndrome, visual analog scale (VAS), and Lysholm scores were determined before treatment, at week 4 and week 8 of treatment, and week 16 of follow-up. Serum interleukin-6 (IL-6), matrix metalloproteinase-3 (MMP-3), tumor necrosis factor-α (TNF-α), osteoprotegerin (OPG), bone gla protein(BGP), and cartilage oligomeric matrix protein (COMP) levels were compared between the two groups before and after 8 weeks of treatment. The clinical efficacy and safety indicators between the two groups were also compared.
Results:
There was no statistical difference in baseline data such as gender, age, disease duration, and body mass index between the two groups of patients. Compared with before treatment, both groups showed decreased WOMAC total and subscale, TCM syndrome total score and symptom scores, and VAS scores, and an increase in Lysholm scores at 4, 8, and 16 weeks after treatment. After treatment, serum IL-6, MMP-3, TNF-α, and COMP levels decreased, whereas serum OPG and BGP levels increased (P<0.05). Compared with the control group, patients in the experimental group showed decreased WOMAC total and subscale, TCM syndrome total score and symptom scores, and VAS scores, and an increase in Lysholm score at 4, 8, and 16 weeks after treatment. Compared with the control group, patients in the experimental group showed decreased serum IL-6, MMP-3, TNF-α, and COMP levels and an increase in serum OPG and BPG levels after treatment (P<0.05). The total effective rate of the experimental group was 91.94%, higher than that of the control group (81.97%; P<0.05).
Conclusion
PRP injection combined with warm acupuncture and moxibustion can improve various TCM symptoms, improve knee joint function and bone metabolism, and reduce inflammation in patients with knee osteoarthritis and cold dampness obstruction syndrome.


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