1.Effect and mechanism of Wnt5a knockdown on the efficacy of M1 bone marrow-derived macrophage in treatment of liver cirrhosis
Feifei XING ; Danyang WANG ; Xinrui ZHENG ; Yannan XU ; Shihao ZHANG ; Junyi ZHAN ; Wei LIU ; Gaofeng CHEN ; Jiamei CHEN ; Ping LIU ; Yongping MU
Journal of Clinical Hepatology 2026;42(3):618-628
ObjectiveTo observe the effect of M1 bone marrow-derived macrophages (M1-BMDM) with Wnt5a knockdown on liver fibrosis and regeneration in a rat model of liver cirrhosis, and to investigate its gain-of-function effect compared with unmodified M1-BMDM. MethodsPrimary bone marrow-derived macrophages were isolated from rats and were polarized to M1 phenotype to construct M1-BMDMWnt5a-KD cells. A rat model of liver cirrhosis induced by CCl4/2-AAF was established, and at the end of week 8, rats were randomly divided into model group, M1-BMDM group, M1-BMDM Wnt5a-knockdown empty vector group (M1-BMDMKD-EV group), and M1-BMDM Wnt5a-knockdown group (M1-BMDMWnt5a-KD group), with 6 rats in each group. On the first day of week 9, the rats in each group were given a single injection of the corresponding cells via the caudal vein, along with an intraperitoneal injection of a CCR2 inhibitor. Six rats without any treatment were used as normal control group. Samples were collected at the end of week 12 to assess liver histopathology, serum liver function parameters, hepatic stellate cell activation, and the expression levels of mature hepatocyte markers. A one-way analysis of variance was used for comparison of continuous data between multiple groups, and the least significant difference t-test was used for further comparison between two groups. ResultsCompared with the model group, all cell treatment groups had significant alleviation of liver inflammatory response and significant reductions in the activities of alanine aminotransferase and aspartate aminotransferase (AST) in serum (all P<0.01), and the M1-BMDMWnt5a-KD group had a significantly lower serum level of AST than the M1-BMDM group (P<0.05). The semi-quantitative analysis based on immunohistochemical staining showed that compared with the model group, all cell treatment groups had a significant reduction in the percentage of CD68-positive area (all P<0.05), and compared with the M1-BMDMKD-EV group, the M1-BMDMWnt5a-KD group had a significant reduction in the percentage of CD68-positive area and a significant increase in the percentage of CD163-positive area (both P<0.05). Compared with the model group, all cell treatment groups had significant reductions in the mRNA expression levels of CD68 and tumor necrosis factor-α (all P<0.05) and the protein expression level of CD68 (all P<0.01); compared with the M1-BMDMKD-EV group, the M1-BMDMWnt5a-KD group had significant increases in the protein and mRNA expression levels of CD163 (both P<0.05), significant reductions in the protein and mRNA expression levels of CD68 (both P<0.05), and a significant reduction in the protein expression level of tumor necrosis factor-α (P<0.01). Sirius Red collagen staining and alpha-smooth muscle actin (α-SMA) immunohistochemical staining showed that compared with the model group, all cell treatment groups had significant alleviation of liver collagen deposition and α-SMA-positive area, with the most significant changes in the M1-BMDMWnt5a-KD group, and compared with the M1-BMDMKD-EV group, the M1-BMDMWnt5a-KD group had significantly smaller Sirius Red-positive area and α-SMA-positive area and a significantly lower content of hydroxyproline in liver tissue (all P<0.05). Compared with the M1-BMDMKD-EV group, the M1-BMDMWnt5a-KD group had significant reductions in the protein and mRNA expression levels of α-SMA and the mRNA expression level of COL-I and TGF-β (all P<0.05). Compared with the model group, all cell treatment groups had a significant increase in the protein expression level of HNF-4α in liver tissue (all P<0.05), and the M1-BMDMWnt5a-KD group had significantly higher protein and mRNA expression levels of HNF-4α and hepatocyte specific antigen than the M1-BMDMKD-EV group (both P<0.05). The M1-BMDMWnt5a-KD group had a significantly higher serum level of albumin than the M1-BMDMKD-EV group (P<0.01). Immunofluorescence co-staining showed that compared with the model group, all cell treatment groups had a significant increase in the number of cells stained positive for HNF and HNF-4α and Ki67 (all P<0.01), and the M1-BMDMWnt5a-KD group had a significantly higher number of such cells than the M1-BMDMKD-EV group (P<0.05). ConclusionInhibition of Wnt5a expression enhances the therapeutic effect of M1-BMDM on rats with liver cirrhosis induced by CCl4/2-AAF, which provides new ideas for enhancing the anti-cirrhotic effect of M1-BMDM through genetic modification.
2.Zuogui Wan Improve Ovarian Inflammatory Microenvironment and Stemness of Ovarian Germline Stem Cells in Ovarian Aging via cGAS/STING Signaling Pathway
Yunling ZHENG ; Xinyi PAN ; Zuang LI ; Yixuan WANG ; Junyi AN ; Yuxin ZOU ; Mengting XIAO ; Zheng CHEN ; Ling ZHU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(7):1-10
ObjectiveTo investigate the mechanism of Zuogui Wan (ZGW) in improving ovarian inflammatory microenvironment and stemness of ovarian germline stem cells (OSCs) for treating ovarian aging via the cyclic guanosine monophosphate/adenosine monophosphate synthase (cGAS)/stimulator of interferon genes (STING) signaling pathway. MethodsForty C57BL/6 female mice were randomly divided into a blank group, a model group, a low-dose ZGW group (2.7 g·kg-1), a high-dose ZGW group (5.4 g·kg-1), and an estradiol valerate group (0.15 mg·kg-1), with 8 mice in each group. Except the blank group, all other groups received a single intraperitoneal injection of cyclophosphamide at 120 mg·kg-1 to establish an ovarian aging mouse model. After successful modeling, each group was continuously administered for 4 weeks, once daily. The physiological status of the mice was observed, and the ovarian index was calculated. The estrus cycle of the mice was monitored. Hematoxylin-eosin (HE) staining was used to observe pathological changes in ovarian tissue. Enzyme-linked immunosorbent assay (ELISA) was used to detect serum sex hormone levels. Serum inflammatory factors interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), and mouse interleukin-6 (IL-6) levels were detected using kits. Western blot was used to detect the protein expression of ovarian cGAS, STING, p-STING, TANK-binding kinase 1 (TBK1), p-TBK1, interferon-induced transmembrane protein 3 (Fragilis), and Vasa homolog protein (MVH). Quantitative real-time polymerase chain reaction (Real-time PCR) was used to detect the mRNA expression of inflammatory factors in ovarian tissue. Immunofluorescence double labeling was performed to locate OSCs in ovarian tissues, and fluorescence intensities of OSCs markers MVH and octamer binding transcription factor 4 (Oct4) were calculated. ResultsCompared with the blank group, the model group showed reduced body weight, ovarian wet weight, and ovarian index (P<0.01) and a disordered estrus cycle (P<0.01). In addition, the levels of serum follicle-stimulating hormone (FSH), TNF-α, IL-6, and IL-1β were increased (P<0.01), while anti-Müllerian hormone (AMH) and estradiol (E2) levels were decreased (P<0.01). The protein expression of cGAS, p-STING/STING, and p-TBK1/TBK1 in ovarian tissue was increased (P<0.05, P<0.01), while that of OSCs stemness factors MVH and Fragilis was reduced (P<0.01). Immunofluorescence indicated a reduction in MVH and Oct4 expression in OSCs (P<0.01). The mRNA expression of inflammatory factors TNF-α, IL-6, and IL-1β in ovarian tissue was increased (P<0.05, P<0.01). Compared with the model group, the treatment groups exhibited improved body weight, ovarian wet weight, and ovarian index (P<0.05) and a reduced rate of estrus cycle disorder (P<0.05, P<0.01). The levels of serum FSH, TNF-α, IL-6, and IL-1β were decreased (P<0.05, P<0.01), while AMH and E2 levels were increased (P<0.01). The protein expression levels of cGAS, p-STING/STING, and p-TBK1/TBK1 in ovarian tissue were decreased (P<0.05), while the protein expression of MVH and Fragilis was increased (P<0.05), and the fluorescence intensities of MVH and Oct4 were increased (P<0.05, P<0.01). The mRNA expression of inflammatory factors in ovarian tissue was decreased (P<0.05). ConclusionZGW alleviate ovarian inflammatory response, regulate ovarian microenvironment homeostasis, and maintain stemness of OSCs in ovarian aging mice probably by modulating the cGAS-STING signaling pathway, thereby improving ovarian function and delaying ovarian aging.
3.A progressive discovery strategy for anti-Alzheimer’s drug identification: In vitro screening with an LPS-induced neuroinflammatory cell model and in vivo validation in 5xFAD mice
Junyi ZHOU ; Linna WANG ; Lianmei WANG ; Pengyue LI ; Yan ZHANG ; Yuheng WANG ; Yan RONG ; Lihua CHEN ; Huijun WANG ; Hongping HOU ; Xiaolu WEI ; Haiyu ZHAO
Science of Traditional Chinese Medicine 2026;4(3):255-269
Background: Alzheimer’s disease (AD) is the most prevalent neurodegenerative disorder. Cellular drug screening represents a key approach in the preclinical discovery of AD therapeutics. Moreover, selecting an appropriate cellular model is essential for exploring the mechanisms underlying AD pathogenesis. Objective: This study aimed to establish a progressive strategy integrating cell model construction, monomer evaluation, and effective component group-based drug discovery for the development of novel therapeutics targeting AD. Methods: A novel AD12 cell line stably expressing the double-transgenic genes amyloid precursor protein (APP)695/presenilin 1(PS1)-ΔE9 was first constructed and subjected to lipopolysaccharide (LPS) stimulation. The resulting LPS-induced neuroinflammatory response was characterized using metabolomic analysis. Subsequently, 16 alkaloids derived from traditional Chinese medicine(TCM) were screened with the LPS-AD12 cell model. Molecular docking was then performed to assess the binding affinity of benzoylaconitine (BAC) with APP and PS1 proteins. A targeted metabolomics approach focusing on amino acids and the tricarboxylic acid (TCA) cycle was applied to BAC-treated LPS-AD12 cells. Finally, aconite extract was administered to five-familial-AD-mutations(5xFAD) transgenic mice, and therapeutic efficacy was evaluated through phenotypic and metabolomic analyses. Results: LPS stimulation induced neuroinflammation in AD12 cells, characterized by elevated levels of interleukin-6, amyloid-β
(Aβ
), and PS1. The primary metabolic pathways affected in AD12 cells included alanine, aspartate, and glutamate metabolism, as well as arginine biosynthesis. Notably, BAC significantly reduced the levels of Aβ
and interleukin-6 in LPS-AD12 cells. Furthermore, molecular docking analyses revealed a strong binding affinity of BAC with APP and PS1 proteins. Targeted metabolomics analysis indicated that the pharmacological activity of BAC was primarily associated with its anti-inflammatory effects, modulation of the cholinergic signaling pathway, and regulation of energy metabolism. Behavioral experiments in 5xFAD mice, together with immunofluorescence assessments of brain tissues (amyloid-β, glial fibrillary acidic protein, and ionized calcium-binding adapter molecule 1), demonstrated that aconite extract treatment significantly alleviated AD-related symptoms. In addition, metabolomics-based mechanistic analyses revealed that serine, acetylcholine, ornithine, and gamma-aminobutyric acid exhibited consistent trends in both brain tissue and serum samples from treated mice. Conclusion: This study established a novel AD12 cell model and an effective, reliable LPS-induced cell-based screening system. Additionally, we proposed a new drug discovery strategy integrating cell model construction, monomer evaluation, and effective component group-based drug discovery. This screening system has the potential to accelerate the identification of anti-AD agents and provides a feasible framework for subsequent in-depth investigations into new drug development from TCM, thereby contributing to the modernization of TCM-derived therapeutics.
4.Analysis of common statistical problems in blood transfusion medical research papers
Junyi CHEN ; Wen WANG ; Zhikai ZHANG
Chinese Journal of Blood Transfusion 2025;38(2):263-267
High-quality scientific research papers need the support of correct and rigorous statistical methods. However, many papers in transfusion medicine research have problems with the improper use of statistical methods. This paper starts from the transfusion medicine related papers published in China in recent years, summarizes and analyzes common problems in the use of statistics from the aspects of research data type description, statistical method selection, statistical result interpretation and statistical content writing, and discusses possible solutions to provide certain references for the writing of research papers in transfusion medicine.
5.Development of a pretreatment workstation for detecting free silica levels in dust
Jian WU ; Yuqiao ZHENG ; Meng LUO ; Mengping ZHANG ; Junyi HUANG ; Fei SHEN ; Feng ZHANG ; Sheng FU ; Xuelei CHEN ; Zongli HUO ; Banghua WU
China Occupational Medicine 2025;52(4):455-459
Objective To investigate an automated pretreatment technology for detecting levels of free silica in workplace dust. Methods An fully automated pretreatment workstation for detecting free silica levels in workplace dust was developed by integrating graphite-controlled digestion temperature, online-controlled dilution of digestion solutions, and filtration endpoint recognition based on monitoring technology, combined with multi-channel synchronous measurements. Results The fully automatic pretreatment workstation was used to digest and filter 14 standard samples of free silica produced by three institutions, and then detected by pyrophosphate method. The result range of high-, medium-, and low-level free silica standard samples detection was 66.5%-84.8%, 40.0%-44.5%, and 2.1%-24.8%, respectively. The mean relative standard deviations were 3.9%, 1.4% and 1.5%. Conclusion The fully automated pretreatment workstation produced results that met relevant requirements. It can effectively replace the manual digestion and filtration steps of the pyrophosphate method to measure free silica levels in workplace dust and enable rapid detection of free silica in dust samples.
6.Application of Anti-tumor Compatibility Structure of Chinese Medicine
Lanpin CHEN ; Feng TAN ; Xiaoman WEI ; Junyi WANG ; Liu LI ; Mianhua WU ; Haibo CHENG ; Dongdong SUN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(8):198-208
Malignant tumors are one of the major diseases that endanger human life and health. Chinese medicine has unique advantages in clinical anti-tumor treatment. However, how to translate the anti-tumor effects of Chinese medicine into clinical practice is the core issue that must be addressed in the process of treating malignant tumors with traditional Chinese medicine (TCM). Unlike modern chemical drugs, the compatibility application of Chinese medicine is the key factor that determines whether Chinese medicine can achieve optimal anti-tumor efficacy and realize the goal of "enhancing efficacy and reducing toxicity". The formulation structure based on this compatibility is the basic form for the safe, efficient, and rational clinical use of anti-tumor Chinese medicine, and it mainly includes three categories: herb pairs, tri-herbal combinations, and compound compatibility. Although herb pairs have the characteristics of a simple structure and strong targeting (enhancing efficacy and reducing toxicity), they often have a single effect and cannot fully address the complex pathogenesis of tumors. As a result, herb pairs are rarely used alone in practice. Compared to herb pairs, tri-herbal combinations broaden the application scope of herbs in clinical treatment, but their therapeutic range remains limited. The traditional "sovereign, minister, assistant, and guide" compound prescription, which includes herb pairs and tri-herbal combinations, improves the efficacy of herbs in treating serious diseases, hypochondriasis, chronic diseases, and miscellaneous disorders. However, due to the limitations of its historical background, it has not been integrated with modern clinical practice and modern pharmacological research, which restricts the development of compound compatibility theory. With the emergence of modern medical technology, it has been combined with traditional compatibility theory of Chinese medicine to create an innovative modern compatibility theory. This includes the "aid medicine" theory derived from modern Chinese medicine pharmacology, which compensates for the inability of the "sovereign, minister, assistant, and guide" theory to accurately apply medicine. Additionally, the "state-targeted treatment based on syndrome differentiation" theory, developed from pharmacology and modern medicine, addresses the deficiency in disease cognition in the "sovereign, minister, assistant, and guide" theory. Under the guidance of these compatibility forms and theories, clinical anti-tumor Chinese medicine can exert its maximum anti-tumor efficacy, which is of great significance for the application of Chinese medicine in clinical tumor treatment.
7.Rh blood group phenotype distribution in some ethnic groups in China: a meta-analysis
Junyi CHEN ; Mengdan SONG ; Jin MA ; Yongyi YANG ; Xiaojuan LI
Chinese Journal of Blood Transfusion 2025;38(4):562-571
[Objective] To analyze the distribution of Rh blood group phenotype in some ethnic groups in China, so as to provide references for accurate blood transfusion. [Methods] The data of CNKI, Wanfang data and VIP were retrieved using "Rh blood group" and "nationality", and the search of PubMed database was conducted with the keywords "Rh blood group", "nationalities", "ethnic groups" and "China", with retrieval time until September 19, 2024 Data were extracted from eligible studies and the literature quality was evaluated using the criteria for cross-sectional studies in STROBE statement. Meta analysis was performed using Stata 11.0 software. [Results] A total of 350 relevant literature were retrieved, of which 26 were included. The total sample size for Rh phenotype distribution detection were 31 432, and the total population for RhD negative screening was 47 227, covering 26 ethnic groups. Meta-analysis revealed that the Rh blood groups phenotype distribution in certain ethnic populations in China was mainly CCDee 46.7% (95%CI=46.2%-47.2%), CcDEe 30.1% (95%CI=29.5%-30.6%), and CcDee 9.0% (95%CI=8.7%-9.3%). Analysis of the RhD-negative phenotype indicated an negative rate of RhD of 0.3% (95%CI=0.2%-0.3%), with the main phenotype distributions of ccdee at 0.2% (95%CI=0.1%-0.2%) and ccdEe at 0.2% (95%CI=0.0%-0.4%). The meta-analysis results of the distribution of common phenotypes among different ethnic groups showed that the CCDee phenotype was mainly distributed as Hani>Dong>Buyi>Miao>Tujia>Hui>Zang>Kazakh>Mongol>Uygur; the CcDEe phenotype: Zang>Mongol>Hui; the CcDee phenotype: Uygur>Kazakh>Mongol>Zang>Hui>Dong>Miao>Tujia>Buyi; the ccDEE phenotype: Zang>Hui=Mongol. The results of this study are similar to those of Qingdao population in China, but differ from studies conducted in North India, German individuals of European ancestry and Saudi Arabian populations. [Conclusion] The distribution of Rh blood group phenotypes in some ethnic groups in China shows no significant difference compared to the Han population, but there are differences when compared to populations in other countries and regions.
8.Monte Carlo simulation study of the effect of filter on radiotherapy dosimetry in superficial X-ray therapy apparatus
Li TAO ; Hui ZHANG ; Yikai WU ; Junyi LIU ; Miao QI ; Ning GAO ; Yankui CHANG ; Xi PEI ; Zhi CHEN ; Xie XU
Chinese Journal of Radiological Medicine and Protection 2025;45(3):194-201
Objective:To explore the dosimetry optimization strategy based on filter thickness and shape selection for the bulb superficial X-ray radiotherapy unit.Methods:Monte Carlo code TOPAS was used to model tubular equipment, and the dose distribution from six X-ray energies (50-150 kV) and five conventional aluminum filters (0.5-3.0 mm) with different thickness were simulated in the water model. The percentage depth dose (PDD) curve along the central axis, the center-axis profile dose at different depths, and the lateral dose distribution were analyzed. The dose distribution of three different designs of aluminum filters (conventional cylindrical, conical and oblique cylindrical filters) was compared to evaluate the effect of dosimetric optimization of different filter shapes.Results:Under the same energy, increasing the thickness of the filter can optimize the superficial skin dose, and the optimization effect of depth dose uniformity can be increased by 26% at a depth of 5.5 mm at 70 kV energy. The raised, flat and inclined dose distribution modes can be achieved by using conventional cylindrical, conical and inclined aluminum filters.Conclusions:By selecting the appropriate X-ray energy and filter thickness, an ideal dose distribution matching the tumor depth can be achieved. The application of personalized filters is also of great significance for diverse target areas.
9.Experiences of food avoidance behavior in patients with inflammatory bowel disease:a qualitative study
Qingyu WANG ; Meijing ZHOU ; Yang LEI ; Sha LI ; Junyi GU ; Zheng LIN ; Qiugui BIAN ; Jiefeng YANG ; Lichen TANG ; Jiali CHEN ; Hantian CHENG
Chinese Journal of Nursing 2025;60(11):1323-1329
Objective This study applies Roy adaptation theory to deeply explore the experience of food avoid-ance behavior in patients with inflammatory bowel disease(IBD),offering insights for developing dietary management strategies.Methods A descriptive qualitative research method was employed.By purposive sampling,24 IBD pa-tients hospitalized in the gastroenterology department of a tertiary hospital in Nanjing from July 2022 to December 2024 were selected for semi-structured interviews.Data were analyzed using a directed content analysis approach.Results This study identified 4 main themes and 11 sub-themes,encompassing overattribution leading to inappro-priate avoidance(recurrent symptoms triggering overattribution,disease staging triggering inappropriate avoidance),negative self-perception leading management struggles(illness fear diminishing self-efficacy,disease trauma eroding self-identity,knowledge deficiency constraining self-determination),functional impairment intensifying role challenges(role internalization undermining social function,social roles relinquishing dietary management),and external con-straints amplifying practical difficulties(family and friend oversight heightening dietary stress,healthcare gaps foster-ing practical helplessness,traditional beliefs restricting dietary exploration,economic hardship limiting balanced nu-trition).Conclusion The interplay of overattribution,negative self-perception,functional impairment,and external constraints in IBD patients hinders their ability to adapt to disease fluctuations,ensnaring them in the adaptive predicament of food avoidance behavior.Healthcare professionals should comprehensively address these factors by fostering accurate perceptions,enhancing psychological support,guiding effective coping strategies,and optimizing ex-ternal resources,thereby improving patients' overall adaptive capacity and promoting their recovery.
10.Analysis of the drug property and resource characteristics of the Chinese herbal medicines used in the Treatise on the Spleen and Stomach
Jindian CHEN ; Xianyuan HE ; Fanying MENG ; Junyi WANG
China Modern Doctor 2025;63(14):53-55,71
Objective To explore the theory of the drug property and resource characteristics contained in the classic clinical work of traditional Chinese medicine Treatise on the Spleen and Stomach,providing references for the clinical use of medicines.Methods Chinese herbal medicines recorded in the prescriptions of Treatise on the Spleen and Stomach were collected and to establish a corresponding database.The theory of drug property recorded was analyzed,and resource characteristics were analyzed by methods of modern Chinese medicinal resource science.Results A total of 63 prescriptions of Treatise on the Spleen and Stomach were collected,involving 105 Chinese medicines.High frequency Chinese medicine include 16 kinds of drugs such as Gancao,Renshen and Chenpi et al.The warm-natured Chinese medicine were the most common used,accounting for 46.67%,the spicy Chinese medicine accountting for 39.74%,mainly belong to the spleen meridian,accounting for 53.53%,toxic Chinese medicine were used less.Conclusion More than 94%of Chinese medicine contained Treatise on the Spleen and Stomach are still in clinical use,and the application of reinforcing drugs,relieving drugs and Qi-regulating drugs is of great significance to clinical guidance.

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