1.Establishment and Evaluation of an Oxidative Stress Model of Atopic Dermatitis Induced by 2,4-dinitrofluorobenzene
Chang LIU ; Xuesong XIANG ; Huihuang HE ; Xiaoqing CHEN ; Wenhong QIU
Laboratory Animal and Comparative Medicine 2026;46(1):46-54
Objective To establish an oxidative stress mouse model of atopic dermatitis (AD) by applying 2,4-dinitrofluorobenzene (DNFB) to the back and post-auricular skin of KM mice, and to evaluate the regulatory role of the RAGE-NLRP3 axis (receptor for advanced glycation end products-NOD-like receptor family, pyrin domain containing 3 axis) in AD-related oxidative stress, thereby providing a potential therapeutic target for AD treatment. Methods Twenty SPF-grade female KM mice were randomly divided into a control group (Control group) and an experimental group (DNFB group), with 10 mice in each group. Mice in the Control group were treated with an acetone-olive oil vehicle (acetone: olive oil = 3:1) on their back and post-auricular skin. Mice in the DNFB group were treated with 0.5% DNFB (prepared by adding 0.5 g DNFB per 100 mL of acetone-olive oil vehicle) on the same areas, once daily for 14 consecutive days. The severity of skin lesions was scored on days 2, 4, 6, 9, 12, and 14 of treatment. On day 14, scratching behavior and ear thickness were evaluated. Ear swelling was evaluated on the final day by measuring bilateral ear thickness three times with a vernier caliper; the three measurements were averaged. HE staining was used to observe morphological and structural changes of cells in the back skin tissues. The mRNA and protein expression levels of RAGE (receptor for advanced glycation end products) in skin tissues were detected by quantitative real-time PCR, Western blot, and immunohistochemical staining. The mRNA expression levels of oxidative stress-related molecules, including NLRP3 (NOD-like receptor family, pyrin domain containing 3), caspase-1 (cysteine-dependent aspartate-specific protease 1), and IL-1β (Interleukin-1β), were detected by quantitative real-time PCR. Results On day 14, the back skin lesion scores of the Control group and DNFB group were (0.20±0.42) and (9.93±1.30) (P<0.000 1), respectively. Scratching behavior scores were (5.00±2.05) and (49.26±8.49) episodes, respectively (P<0.000 1), and ear thicknesses were (213.00±11.87) μm and (765.93±140.47) μm (P<0.000 1), respectively. The DNFB group exhibited marked skin dryness, desquamation, and thickening. HE staining results showed that skin inflammation was obvious in the DNFB group, consistent with the pathological features of AD. Quantitative real-time PCR and Western blot results showed that compared with the Control group, the mRNA expression level of RAGE in skin tissues of the DNFB group was significantly increased (P<0.05), and the protein expression level of RAGE was also significantly increased (P<0.01). Immunohistochemical staining results showed that compared with the Control group, skin tissue sections of the DNFB group exhibited thickened stratum corneum and fibrotic proliferation of fibroblasts in the interstitium under microscopic observation, with a significant increase in RAGE protein expression in the skin tissues (P<0.01). Quantitative real-time PCR results showed that the mRNA expression levels of NLRP3, caspase-1, and IL-1β in skin tissues of the DNFB group were all significantly increased (P<0.01). Conclusion The AD mouse oxidative stress model has been successfully established by topical DNFB application. RAGE may promote the development of AD by regulating the NLRP3 inflammasome and IL-1β release, forming an oxidative-inflammatory cascade, suggesting that it could be a potential therapeutic target for AD.
2.Association between Mediterranean diet scores and dental caries among children and adolescents with neurodevelopmental disorders
XIONG Wenjuan, SU Yuanyuan, LIU Zhao, HUANG Xiaoqing, QU Zhiyi, CUI Shanshan
Chinese Journal of School Health 2025;46(2):172-176
Objective:
To explore the association between mediterranean diet (MD) patterns and dental caries among children and adolescents with neurodevelopmental disorders (NDD), so as to provide a basis for developing scientific anti caries strategies related to diet.
Methods:
From December 2021 to June 2024, a questionnaire survey, a three day 24 hour dietary review survey, oral health examination, physical development measurement and Childhood Autism Rating Scale (CARS) evaluation were conducted involving 147 children and adolescents aged 2-22 years with NDD from nine special education schools and rehabilitation institutions in Tianjin. Group comparisons were carried out using the Mann-Whitney U test, Chi-square test, or Fisher s exact probability method. The correlation between dietary quality and dental caries was analyzed by adopting multiple linear regression analysis and restricted cubic spline.
Results:
There were 46 children and adolescents (31.3%) in the non dental caries group and 101 children and adolescents (68.7%) in the dental caries group. The number of decayed missing and filled teeth (dmft) was 2.0 (4.0), and the MD score was 4.0 (2.0) points. There were 62 children and adolescents (42.2%) in the low MD scores group and 85 children and adolescents (57.8%) in the high MD scores group. There was no significant difference in MD scores between NDD children in the non dental caries group and those in the dental caries group [nondental caries group:4.0(2.0), dental caries group:4.0(2.0), Z= -0.14, P >0.05]. The MD scores and dmft exhibited increasing and then decreasing trend ( P total =0.02, P non lineary = 0.04 ). Children and adolescents with NDD in the MD high scores group had a lower number of dmft than those in the MD low scores group ( β= -2.00 , 95%CI =-3.39 to -0.62, P <0.05). However, in children and adolescents with NDD and CARS scores ≥30, the above association was insignificant ( β=-0.63, 95%CI=-0.29-0.15, P >0.05).
Conclusion
Children and adolescents with NDD who have dietary patterns similar to the Mediterranean diet, are found to have fewer dental caries, and this is observed among those with no or mild symptoms of autism spectrum disorder.
3.Literature Based Analysis on Adverse Reactions in Simultaneously Clinical Use of Banxia (Rhizoma Pinelliae)- Wutou (Aconitum)
Can CAO ; Wenyong LIAO ; Jiwen ZHANG ; Yinghao WU ; Xiangnan XU ; Meijing WU ; Xiaoqing LIU ; Shaohong CHEN ; Haiyan LIU ; Linlin XIU ; Xiangqing CUI ; Gaoyang LI ; Ying ZHANG ; Gansheng ZHONG
Journal of Traditional Chinese Medicine 2025;66(9):955-962
ObjectiveTo analyze the adverse reactions associated with the clinical use of Banxia (Rhizoma Pinelliae)- Wutou (Aconitum) in the same formula, with the aim of providing a reference for the safety of their clinical application. MethodsLiterature on the clinical application of antagonistic herbs "Banxia-Wutou" used in the same formula, published from January 1st, 2014, to June 30th, 2023, was retrieved from databases including CNKI, VIP, Wanfang, SinoMed, PubMed, Cochrane Library, and Embase. A database was established, and information related to adverse reactions was extracted, including descriptions, classifications, specific manifestations, management and outcomes, patients' primary diseases (western medicine diseases and traditional Chinese medicine diagnoses and syndromes), and medication information (dosage, ratio, administration routes, and dosage forms). ResultsA total of 79 researches simultaneously used antagonistic herbs Banxia-Wutou in the same formula and reported associated advers reactions. Gastrointestinal adverse reactions were the most common, with 8 studies reporting management of adverse reactions and 3 studies reporting improvement with no intervention. Among the 11 researches, the adverse reaction relieved to extant, while other 69 researches didn't report the managment of adverse reaction and its prognosis. For the primary disease in western medicine system, chronic bronchitis and chronic obstructive pulmonary disease (COPD) were most common, while gastric pain was the most common symptom in traditional Chinese medicine with spleen and kidney deficiency and spleen stomach cold deficiency being the most frequent syndromes. The most common Banxia dosage was 10 g, while for the Wutou, Fuzi (Radix Aconiti Lateralis Praeparata) was predominant with the highest dose at 15 g. The most frequent herbal combination was Banxia-fuzi, with a 1∶1 ratio. The main administration route was oral, and the primary dosage form was decoction. ConclusionGastrointestinal adverse reactions are the most common in the clinical use of Banxia-Wutou antagonistic herb combinations. Research on the safety of "Banxia-Wutou" combinations should focus on respiratory system diseases and spleen-stomach related conditions.
4.Analysis of Tongue Coating Microbiota Characteristics in Coronary Heart Disease with Qi Deficiency and Blood Stasis Syndrome
Chuhao WANG ; Yongyue LIU ; Zhaoxuan DING ; Xiaoqing ZHANG
Journal of Traditional Chinese Medicine 2025;66(5):501-508
ObjectiveTo explore the characteristics of the tongue coating microbiota in patients of coronary heart disease (CHD) with qi deficiency and blood stasis syndrome. MethodsA total of 27 CHD patients with qi deficiency and blood stasis syndrome, 29 patients with non-qi deficiency and blood stasis syndrome, and 20 healthy individuals were included in this study. The tongue coating microbiota of the participants was analyzed using 16S rDNA high-throughput sequencing technology, followed by Alpha and Beta diversity analyses and comparisons of microbial abundance. ResultsA total of 479 operational taxonomic units (OTUs) were detected, among which 245 OTUs were shared across all three groups. There were 33 OTUs unique to the qi deficiency and blood stasis syndrome group, 21 OTUs unique to the non-qi deficiency and blood stasis syndrome group, and 121 OTUs unique to the healthy group. The observed species count (Sob), total species richness (Chao1), abundance-based coverage estimator (ACE), and Shannon diversity index were significantly lower in the qi deficiency and blood stasis syndrome and non-qi deficiency and blood stasis syndrome groups compared to the healthy group (P<0.05). Principal coordinate analysis (PCoA) of the tongue coating microbiota showed significant differences in distance matrices among the three groups (P<0.05). Compared with the healthy group, the qi deficiency and blood stasis syndrome group exhibited an increased abundance of Actinobacteria, Patescibacteria, Spirochaetes, Verrucomicrobia, Rothia, TM7X, Gemella, and Corynebacterium, while Fusobacteria, Cyanobacteria, Leptotrichia, and Lactobacillus decreased (P<0.05). In the non-qi deficiency and blood stasis syndrome group, Actinobacteria, Verrucomicrobia, Rothia, and Corynebacterium increased, whereas Cyanobacteria and Lactobacillus reduced (P<0.05). When comparing with the non-qi deficiency and blood stasis syndrome group, the qi deficiency and blood stasis syndrome group had a significantly higher abundance of Patescibacteria, Peptostreptococcus, Solobacterium, Filifactor, Moraxella, Porphyromonas endodontalis, and Capnocytophaga, while Cyanobacteria reduced (P<0.05). Conclusuion Patients with CHD of qi deficiency and blood stasis syndrome exhibit a decrease in beneficial bacteria and an increase in pathogenic bacteria. Patescibacteria, Peptostreptococcus, Solobacterium, Filifactor, Moraxella, Porphyromonas endodontalis, and Capnocytophaga were identified as the key differential microbiota distinguishing qi deficiency and blood stasis syndrome from non-qi deficiency and blood stasis syndrome patients.
5.Effect of Zishen Tongguan Formula on "Gut-prostate" Axis of Rats with Chronic Non-bacterial Prostatitis Based on 16S rDNA Sequencing
Xiran LI ; Mengjiao CHEN ; Kaiping ZOU ; Chenguang ZHAO ; Xingbin DAI ; Xiaoqing ZHANG ; Shun LIU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(7):63-71
ObjectiveBased on the theory of "gut-prostate" axis, this study explored the effects and mechanisms of Zishen Tongguan formula and Cinnamomi Cortex in the formula in treating rats with chronic non-bacterial prostatitis(CNP) by detecting the levels of inflammatory factors, and the composition and structure of intestinal flora in CNP rats. MethodsEight out of 42 SD rats were randomly selected as the normal group, and the remaining rats were injected with carrageenan to prepare the CNP model. After successful modeling, 32 rats were randomly divided into the model group, Ningmitai capsule group(0.50 g·kg-1), Zishen Tongguan formula group(2.00 g·kg-1), and the Phellodendri Chinensis Cortex-Anemarrhenae Rhizoma pair group(PCC-AR group, 2.00 g·kg-1), with 8 rats in each group. The administered groups were given the corresponding medicinal solution by gavage, and the normal and model groups were intragastrically administered with an equal volume of normal saline, once a day for 14 consecutive days. The prostate tissues of rats were collected and subjected to hematoxylin-eosin(HE) staining and Masson staining to observe the pathological changes of the tissues in each group. Enzyme-linked immunosorbent assay(ELISA) was used to detect the levels of related inflammatory factors in rat serum, and 16S rDNA sequencing was used to analyze the abundance and diversity changes of gut microbiota before and after administration, and species difference analysis was performed. ResultsAll the administered groups could alleviate the inflammatory symptoms of CNP rats, increase the expression levels of anti-inflammatory factors and decrease the expression levels of pro-inflammatory factors, with the most sIgnificant effect observed in the Zishen Tongguan formula group. Compared with the normal group, the expression levels of interleukin(IL)-8, hypersensitive C-reactive protein(hs-CRP), immunoglobulin(Ig)M, secretory IgA (sIgA), and inducible nitric oxide synthase(iNOS) were sIgnificantly increased in the model group(P<0.01). Compared with the model group, the expression levels of the above inflammatory factors in all administered groups were significantly reduced(P<0.01). When compared with the PCC-AR group, the Zishen Tongguan formula group showed a significant decrease in transforming growth factor(TGF)-β1 expression level(P<0.05) and a significant increase in IgM expression level(P<0.01). The results of gut microbiota analysis showed that, compared with the PCC-AR group, at the order level, the Zishen Tongguan formula group significantly reduced the relative abundance of conditional pathogens such as Bacteroidales, Acidaminococcales, Rhodospirillales, Clostridiales, and Elusimicrobiales(P<0.01). And at the genus level, the Zishen Tongguan formula group significantly decreased the relative abundance of pathogenic microbiota such as Lachnospira and Bacteroides(P<0.01) and significantly increased the relative abundances of beneficial microbiota such as Ruminococcus and Lactobacillus(P<0.01). ConclusionZishen Tongguan formula can reduce the level of harmful intestinal bacteria, increase the level of beneficial intestinal bacteria, down-regulate the expression of serum inflammatory factors, and the small amount of Cinnamomi Cortex in the formula may play a key role in the treatment of CNP with this formula.
6.Effect of Zishen Tongguan Formula on "Gut-prostate" Axis of Rats with Chronic Non-bacterial Prostatitis Based on 16S rDNA Sequencing
Xiran LI ; Mengjiao CHEN ; Kaiping ZOU ; Chenguang ZHAO ; Xingbin DAI ; Xiaoqing ZHANG ; Shun LIU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(7):63-71
ObjectiveBased on the theory of "gut-prostate" axis, this study explored the effects and mechanisms of Zishen Tongguan formula and Cinnamomi Cortex in the formula in treating rats with chronic non-bacterial prostatitis(CNP) by detecting the levels of inflammatory factors, and the composition and structure of intestinal flora in CNP rats. MethodsEight out of 42 SD rats were randomly selected as the normal group, and the remaining rats were injected with carrageenan to prepare the CNP model. After successful modeling, 32 rats were randomly divided into the model group, Ningmitai capsule group(0.50 g·kg-1), Zishen Tongguan formula group(2.00 g·kg-1), and the Phellodendri Chinensis Cortex-Anemarrhenae Rhizoma pair group(PCC-AR group, 2.00 g·kg-1), with 8 rats in each group. The administered groups were given the corresponding medicinal solution by gavage, and the normal and model groups were intragastrically administered with an equal volume of normal saline, once a day for 14 consecutive days. The prostate tissues of rats were collected and subjected to hematoxylin-eosin(HE) staining and Masson staining to observe the pathological changes of the tissues in each group. Enzyme-linked immunosorbent assay(ELISA) was used to detect the levels of related inflammatory factors in rat serum, and 16S rDNA sequencing was used to analyze the abundance and diversity changes of gut microbiota before and after administration, and species difference analysis was performed. ResultsAll the administered groups could alleviate the inflammatory symptoms of CNP rats, increase the expression levels of anti-inflammatory factors and decrease the expression levels of pro-inflammatory factors, with the most sIgnificant effect observed in the Zishen Tongguan formula group. Compared with the normal group, the expression levels of interleukin(IL)-8, hypersensitive C-reactive protein(hs-CRP), immunoglobulin(Ig)M, secretory IgA (sIgA), and inducible nitric oxide synthase(iNOS) were sIgnificantly increased in the model group(P<0.01). Compared with the model group, the expression levels of the above inflammatory factors in all administered groups were significantly reduced(P<0.01). When compared with the PCC-AR group, the Zishen Tongguan formula group showed a significant decrease in transforming growth factor(TGF)-β1 expression level(P<0.05) and a significant increase in IgM expression level(P<0.01). The results of gut microbiota analysis showed that, compared with the PCC-AR group, at the order level, the Zishen Tongguan formula group significantly reduced the relative abundance of conditional pathogens such as Bacteroidales, Acidaminococcales, Rhodospirillales, Clostridiales, and Elusimicrobiales(P<0.01). And at the genus level, the Zishen Tongguan formula group significantly decreased the relative abundance of pathogenic microbiota such as Lachnospira and Bacteroides(P<0.01) and significantly increased the relative abundances of beneficial microbiota such as Ruminococcus and Lactobacillus(P<0.01). ConclusionZishen Tongguan formula can reduce the level of harmful intestinal bacteria, increase the level of beneficial intestinal bacteria, down-regulate the expression of serum inflammatory factors, and the small amount of Cinnamomi Cortex in the formula may play a key role in the treatment of CNP with this formula.
7.Expression characteristics of metabotropic glutamate receptor 4 in cochlear hair cells during early postnatal stages in mice
Zhiji CHEN ; Xuelai LIU ; Xiaoqing ZHOU ; Yuting SHE ; Menglong FENG ; Shaojing KUANG ; Hongyan FANG ; Wei YUAN
Journal of Army Medical University 2025;47(11):1227-1234
Objective To investigate the spatiotemporal expression pattern of metabotropic glutamate receptor 4(mGluR4)in the mouse cochlear basilar membrane and its dynamic changes during postnatal development until hair cell synaptic maturation.Methods Ten healthy 3-month-old C57BL/6L wild-type mice(5 males,5 females,25~35 g)were bred to obtain offspring.Cochlear samples from postnatal days 1~30(P1~P30)were analyzed.Frequency-specific auditory brainstem response(ABR)thresholds were measured at 45.2,32.0,22.6,16.0,11.3,8.0,5.6,and 4.0 kHz in P12,P14,P16,P21,and P30 mice.Immunofluorescence assay combined with confocal laser scanning microscopy was used to scan cochlear whole-mount preparations.Spatiotemporal co-localization profiling of mGluR4 and synaptic markers Ctbp2 and Snap25 was observed,and quantitative comparisons of fluorescent puncta density were performed.Results ABR testing revealed undifferentiated waveforms in all P12 mice,while 70%of P14 mice exhibited well-defined Wave I.In P14,P16,P21 and P30,mice undergoing ABR testing,50%failed to exhibit ABR waveforms when stimulated by 45.2 kHz tones,and 87.5%showed no response to 32.0 kHz stimuli.Auditory response thresholds for other frequencies progressively decreased with increasing postnatal age.Immunolocalization demonstrated stronger mGluR4 expression in inner hair cells(IHCs)versus outer hair cells(OHCs).Distinct mGluR4 puncta were observed at IHC presynaptic active zones(co-localized with Snap25)as early as P11,but showed minimal overlap with CtBP2-labeled ribbons.Quantitative analysis revealed significantly higher mGluR4 puncta density at P30 compared to P14 synapses(P<0.05).Conclusion Postnatal upregulation of mGluR4 at IHC synaptic regions correlates with functional maturation.These findings suggest mGluR4 may modulate vesicular exocytosis regulation and exert pharmacological inhibition on glutamate release,potentially influencing synaptic plasticity during auditory pathway refinement.
8.Research progress in transcranial ultrasound stimulation for modulation of Alzheimer′s disease
Xiaoyu SONG ; Xu LIU ; Xiaoqing ZHOU ; Tao YIN ; Zhipeng LIU
International Journal of Biomedical Engineering 2025;48(2):197-204
Transcranial ultrasound stimulation (TUS) is a non-invasive brain stimulation technique that demonstrates great potential in treatment of neurological disorders due to its high spatial resolution and focused penetration ability, enabling millimetre-level precision modulation of specific brain regions with ultrasound to achieve neuromodulation of targeted areas. At present, numerous animal experiments and human studies have been carried out on the regulation of Alzheimer′s disease by TUS. In this review, the regulation effects of TUS on Alzheimer′s disease were introduced from the perspective of targeting different brain regions, including the hippocampus, frontal lobe and whole brain.
9.A brief discussion on TCM diagnosis and treatment of myelodysplastic syndrome based on the Theory of " Sui Qi Suo De"
Yunhe QI ; Haiyan CHEN ; Ming GUO ; Junxia LIU ; Ling LI ; Junyao LIAO ; Jing LIAO ; Xiaoqing DING
International Journal of Traditional Chinese Medicine 2025;47(3):294-297
The theory of " Sui Qi Suo De" originates from Zhang Zhongjing's Jin Gui Yao Lue and has been further developed by later generations of practitioners, offering significant guidance for clinical practice. Myelodysplastic syndromes (MDS) are common malignant disorders of the hematopoietic system, characterized by high heterogeneity and progressive mutational changes. In Traditional Chinese Medicine (TCM), MDS falls under the category of "marrow toxin exhaustion". This article applies the theory of " Sui Qi Suo De" in TCM to analyze the pathophysiological changes during different stages of MDS. Specifically, it explores the precursor stage (focusing on health maintenance and prevention before illness, addressing the " Suo De" of "gradual decline of vital qi"), the low-risk stage (strengthening the spleen and kidneys, clearing toxic pathogens, addressing the " Suo De" of "weakened vital qi invaded by pathogens"), and the medium-to-high-risk stage (detoxifying and reinforcing the body, harmonizing physical and mental health, addressing the " Suo De" of "dominant pathogens and declining vital qi"). The goal is to provide new directions and theoretical insights for the TCM treatment of MDS.
10.Investigation on influencing factors of TCM constitutions among female medical workers in a tertiary TCM Hospital
Xiaoqing CAI ; Yan LI ; Xiaochun CUI ; Xinyi LIU ; Xujie WANG ; Huidong ZHU ; Xiupeng YANG
International Journal of Traditional Chinese Medicine 2025;47(10):1384-1390
Objective:To explore the impact of lifestyle on TCM constitutions among female medical workers at Xiyuan Hospital, Chinese Academy of Traditional Chinese Medicine using questionnaires; To provide references for optimizing their health management.Methods:A cross-sectional study was conducted. A total of 500 female medical workers were randomly selected from Xiyuan Hospital from April to December 2022 to complete the questionnaires. The relationship between lifestyle and constitutions of female medical workers was analyzed.Results:A total of 457 cases completed the questionnaires. Among the 9 constitutions, 102 people (22.32%) had a peaceful constitution, while 355 people (77.68%) had a biased constitution; the top three biased constitutions were qi stagnation constitution 90 people (19.69%), yang deficiency constitution 83 people (18.16%), and phlegm-dampness constitution 56 people (12.25%). The factors that influenced physical constitutions according to the severity of lifestyle were: sleep, diet, mood, exercise, labor, environment, children, and education.Conclusions:Female medical workers have a high proportion of biased constitution, and the main types of biased constitution are qi stagnation constitution, yang deficiency constitution, and phlegm-dampness constitution. According to the existing biased constitution, targeted adjustments and interventions are carried out, and it is of great significance to protect and care for the health of female medical workers from the concept of preventive treatment of diseases with TCM.


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