1.Correlation Analysis Between Microbial Community Changes and Medicinal Quality Formation During Processing of Angelicae Dahuricae Radix
Xiaoyan CHEN ; Xinglong ZHU ; Qingxia GAN ; Jiahao WANG ; Guangqin AN ; Qinghua WU ; Jin PEI ; Yuntong MA
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(4):198-207
ObjectiveTo compare the differences in color, odor, coumarin content and microbial community composition of Angelicae Dahuricae Radix(ADR) during different drying processes, and to explore the correlation between changes in microbial community composition and changes in quality indexes of ADR. MethodsThe fresh ADR was processed at three drying temperatures(50, 70, 100 ℃) by drying and steaming cutting, semi-fresh cutting and drying, fresh cutting and drying, and sulfur fumigation methods. The color values of samples were extracted by Adobe Photoshop 2022 software and subjected to principal component analysis(PCA), electronic nose was used to identify the odor information of medicinal powders and subjected to loadings analysis, PCA, and linear discriminant analysis(LDA), and high performance liquid chromatography(HPLC) was used to determine the contents of five coumarins(bergapten, oxypeucedanin, imperatorin, phellopterin, isoimperatorin). The samples for microbial detection were taken from fresh dried samples, 50 ℃(dried and steamed cut, sulfur fumigated) samples, and 100 ℃(dried and steamed cut) samples when the water content was 50% and 14%, respectively. And the changes of microbial community composition during processing were determined by high-throughput sequencing method. The relationship between the changes of microbial community composition and the changes of odor, color and active component content of ADR during drying process was analyzed by Pearson correlation analysis. ResultsThe color quantification results showed that an increase in drying temperature led to the decrease of brightness value(L), and the increases of red-green value(a) and yellow-blue value(b), and the change of processing method had no obvious effect on the color of medicinal materials. The results of odor quantification showed that W1S, W2S, W5S, W2W and W1W sensor were sensitive to the odor changes of ADR and could be used to distinguish ADR decoction pieces from different processing methods. The results of HPLC showed that the coumarin content of ADR decreased with the increase of drying temperature and the delay of processing time, the optimal processing method was drying and steaming cutting method, and the optimal temperature was 50 ℃. High-throughput sequencing results showed that the dominant bacteria in ADR during processing were Achromobacter, Agrobacterium, Nocardioides, Mycobacterium and Enterobacter, the dominant fungi were Coprinopsis, Meyerozyma and Apiotrichum. The results of correlation analysis showed that the quality indexes of ADR were positively correlated with Agrobacterium, Mycobacterium in bacteria, Candida in fungi, and negatively correlated with Bacillus in bacteria. ConclusionThere are significant differences in the color, odor, coumarin content and microbial community composition of ADR in different drying processes, and the best drying method is drying and steaming cutting at 50 ℃. The relative abundance changes of 9 bacterial genera and 4 fungal genera are closely related to the quality formation of ADR during the drying process.
2.Research on Historical Evolution and Clinical Application of Shengyutang
Yuning CAO ; Wenpan LIU ; Tianyu MA ; Lanru CHEN ; Jiahao HUANG ; Yonghai MENG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(4):235-243
Shengyutang is a famous classical formula of tonic, which is made from Siwutang with Ginseng Radix et Rhizoma and Astragali Radix. It is included in the Catalogue of Ancient Famous Classical Formulas(The First Batch). Based on the Principles of Key Information Research of Ancient Famous Classical Formulas, this paper used bibliometrics to sort out and research the key information of Shengyutang in the aspects of history, composition, origin and processing, dosage, decocting method, efficacy and indications. After research, it has been found that this formula was first recorded in Lanshi Micang written by LI Dongyuan during the Jin dynasty, composed of Rehmanniae Radix, Rehmanniae Radix Praeparata, Chuanxiong Rhizoma, Ginseng Radix et Rhizoma, Astragali Radix and Angelicae Sinensis taproot. The name of the formula passed down through generations was relatively unified, with clear origins and veins. In later generations, this formula was the mainstream, and adjustments were made to the dosage and composition according to the indication. In the formula, Astragalus membranaceus var. mongholicus was selected as the origin of Astragali Radix, and the origins of other medicinal materials were consistent with the 2020 edition of Chinese Pharmacopoeia. Except for Rehmanniae Radix Praeparata, the other medicinal materials were made from raw products, and the dosage form was boiled powder. According to the measurement standard in the Jin dynasty, the recommended usage and dosage were 1.24 g of Rehmanniae Radix, Rehmanniae Radix Praeparata, Chuanxiong Rhizoma, Ginseng Radix et Rhizoma each, 2.07 g of Astragali Radix and Angelicae Sinensis taproot each, crushed into coarse particles that pass through the 4 mesh sieve but can't pass through the 10 mesh sieve, added 1 200 mL of water and boiled to 300 mL, and removed the residue. Shengyutang has the functions of tonifying Qi and blood, and blood intake, treating various sores, and restlessness and insomnia caused by excessive blood flow. In ancient times, this formula was widely used in the treatment of surgical sores, gynecological diseases, deficiency syndrome, etc. In modern clinical practice, it is mostly used to treat gynecological, neurological, musculoskeletal, hematological diseases caused by Qi and blood deficiency. In this paper, the key information of Shengyutang was researched by reviewing relevant ancient literature, in order to provide reference for the modern application and development of this formula.
3.Effect of targeted silencing of DNMT3A on collagen deposition, proliferation and migration activity of mouse lung fibroblasts
Xianchen Wang ; Junbo You ; Hui Ling ; Jiahao Fan ; Qi Chen ; Hui Tao ; Jiming Sha
Acta Universitatis Medicinalis Anhui 2025;60(1):66-72
Objective:
To investigate the effect of targeted silencing of DNA methyltransferase 3A(DNMT3A) on collagen deposition, proliferation and migration activity of mouse lung fibroblasts(PFs).
Methods:
In order to ensure the proliferation and migration activity of primary fibroblasts, the lung tissues of neonatal C57 suckling mice were taken, PFs were extracted after being sheared, and the morphology was observed and identified under the microscope. PFs cells were activated by 5 ng/ml TGF-β1for 24 h after cell attachment, and DNMT3A silencing model was constructed by small interfering RNA; The experiment was divided into control group, TGF-β1group, TGF-β1+ siRNA-NC group and TGF-β1+ siRNA-DNMT3A group. The protein expressions of DNMT3A, α-smooth muscle actin(α-SMA) and Collagen Ⅰ were detected by Western blot; Real time quantitative reverse transcription polymerase chain reaction(RT-qPCR) was used to detect the mRNA expression changes ofDNMT3A,α-SMAandCollagenⅠ. The proliferation ability of PFs was detected by CCK-8 and EdU staining; the migration ability of PFs was detected by scratch test and Transwell migration test.
Results:
Compared with the control group, TGF-β1induced the increase of DNMT3A in the activated PFs cell group(P<0.01), the protein and mRNA levels of fibrosis and proliferation related indicators α-SMA and Collagen Ⅰ also increased(allP<0.05), and the proliferation and migration ability of PFs increased(allP<0.000 1). Compared with the siRNA-NC group, the protein expression levels of DNMT3A(P<0.000 1) and related indicators α-SMA(P<0.01) and Collagen Ⅰ(P<0.01) significantly decreased in the DNMT3A silencing group by Western blot, and the mRNA levels ofDNMT3A,α-SMAandCollagenⅠby RT-qPCR also decreased(allP<0.001), and the proliferation(P<0.01) and migration ability(P<0.05) of PFs cells decreased compared with the control group.
Conclusion
Silencing DNMT3A can inhibit the deposition of collagen and the proliferation of PFs. DNMT3A can promote the proliferation and migration of PFs, and then promote the activation of PFs and the development of pulmonary fibrosis. This process may be regulated by DNA methylation modification.
4.Textural Research on Key Information of Liuhetang
Jiaxin GAO ; Jiahao WANG ; Renshou CHEN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(10):234-244
Liuhetang is one of the classic prescriptions included in the Catalogue of Ancient Classic Prescriptions (the Second Batch). This study adopts the method of literature review to systematically sort out the ancient literature about Liuhetang and obtained a total of 127 effective data records, involving 82 ancient books (including 2 Japanese books). The origin, medicinal composition, compatibility, original plants and their processing methods, dosage, decocting method, usage, and indications of Liuhetang were analyzed. Liuhetang is first recorded in the Formulary of the Bureau of Taiping People's Welfare Pharmacy in the Song Dynasty, consisting of Amomi Fructus, Pinelliae Rhizoma, Armeniacae Semen Amarum, Ginseng Radix et Rhizoma, Glycyrrhizae Radix et Rhizoma, red Poria, Pogostemonis Herba, Lablab Semen Album, Chaenomelis Fructus, Moslae Herba, Magnoliae Officinalis Cortex, Zingiberis Rhizoma Recens, and Jujubae Fructus. The original plants of these herbal medicines follow those in the 2020 edition of the Pharmacopoeia of the People's Republic of China. The raw materials of Amomi Fructus, Armeniacae Semen Amarum, Ginseng Radix et Rhizoma, Glycyrrhizae Radix et Rhizoma, red Poria, Pogostemonis Herba, Chaenomelis Fructus, Moslae Herba, Magnoliae Officinalis Cortex, Zingiberis Rhizoma Recens, and Jujubae Fructus are used in this prescription. Pinelliae Rhizoma, Glycyrrhizae Radix et Rhizoma, Lablab Semen Album, and Magnoliae Officinalis Cortex are processed with alum, stir-fried, processed with Zingiberis Rhizoma Recens, and processed with Zingiberis Rhizoma Recens, respectively. The recommended formula is composed of 0.79 g Amomi Fructus, 0.79 g Pinelliae Rhizoma, 0.79 g Armeniacae Semen Amarum, 0.79 g Ginseng Radix et Rhizoma, 0.79 g Glycyrrhizae Radix et Rhizoma, 1.57 g red Poria, 1.57 g Pogostemonis Herba, 1.57 g Lablab Semen Album, 1.57 g Chaenomelis Fructus, 3.15 g Moslae Herba, and 3.15 g Magnoliae Officinalis Cortex. The above medicines should be pulverized to reach 10 meshes, mixed with 450 mL water, 3 g Zingiberis Rhizoma Recens, and 3 g Jujubae Fructus, and decocted to reach a volume of 240 mL. The filtrate should be taken three times a day. In ancient times, Liuhetang was mainly used to treat cholera, vomiting, diarrhea, phlegm, dyspnea, cough, chest distension, dizziness and pain in the head, swelling in the limbs, lethargy, loss of appetite, difficult urination and dark urine caused by heat and dampness damage to the spleen and disharmony between spleen and stomach. In modern times, Liuhetang is mainly used to treat the digestive system diseases such as gastroenteritis, hepatitis, stomach pain, and diarrhea. The above research confirmed the key information of Liuhetang, providing a basis for the clinical application of this prescription.
5.Combination of effective ingredients of traditional Chinese medicine and bone tissue engineering materials for bone repair
Yaokun WU ; Chenglin LIU ; Jiahao FU ; Wei SONG ; Hao CHEN ; Hongzhong XI ; Xin LIU ; Bin DU ; Guangquan SUN
Chinese Journal of Tissue Engineering Research 2025;29(10):2141-2150
BACKGROUND:How to repair bone defect has been a clinical problem for a long time.The effective ingredients of traditional Chinese medicine have good biological activity and therapeutic effect,and the combination of effective ingredients of traditional Chinese medicine and tissue engineering materials has a broad prospect in the field of bone repair.The combination of different effective ingredients of traditional Chinese medicine and scaffolds has similarities in their functional relationships. OBJECTIVE:To collect the cases of the combinations of effective ingredients of traditional Chinese medicine and scaffolds,then analogize tissue engineering scaffolds and effective ingredients of traditional Chinese medicine into two types of traditional Chinese medicine that generate compatibility relationships based on the inspiration of the compatibility of seven emotions and summarize the relationship between the two based on their functional relationships. METHODS:Relevant articles from January 1998 to January 2024 were searched in PubMed and China National Knowledge Infrastructure(CNKI),using English search terms"traditional Chinese medicine,Chinese medicine,traditional Chinese medicine monomers,bone defect,bone repair,bone tissue engineering,tissue engineering,scaffold"and Chinese search terms"traditional Chinese medicine,effective ingredients of traditional Chinese medicine,traditional Chinese medicine monomers,bone tissue engineering,bone tissue engineering scaffold,scaffold,tissue engineering,bone defect,bone repair."A total of 88 articles were included for review and analysis. RESULTS AND CONCLUSION:(1)Both tissue engineering scaffold materials and active ingredients of traditional Chinese medicine have been widely used in the field of bone repair.Although they have obvious advantages in osteogenesis,there are still many shortcomings.Many studies are dedicated to preparing composite materials from the two,hoping to exert a detoxification and synergism through the interaction between the two.(2)Some drugs and materials can promote each other in osteogenesis,antibacterial,and promoting angiogenesis,enhancing their original effects.Inspired by the traditional concept of prescription compatibility,this article summarized it as a"Mutual promotion"relationship and provided examples to support it.(3)Some drugs can enhance the strength of materials,while some materials can achieve sustained release and controlled release effects,increase drug loading and stability,or achieve targeted delivery of drugs loaded on them.The article summarized this unilateral enhancement effect as a"Mutual assistance"relationship.(4)The combination of some traditional Chinese medicine and materials can reduce the toxic side effects of the other party.The article summarizes this detoxification relationship as"Mutual restraint and detoxification."(5)The article provided a new perspective on traditional Chinese medicine composite scaffolds,inspired by the seven emotions compatibility relationship and based on the classification of action relationships.It introduced traditional Chinese medicine concepts into the field of tissue engineering,providing new research ideas for subsequent researchers of composite scaffolds,and providing certain convenience in material selection and matching.
6.Relationship between long non-coding RNA and osteoarthritis
Shanbin ZHENG ; Tianwei XIA ; Jiahao SUN ; Zhiyuan CHEN ; Xun CAO ; Chao ZHANG ; Jirong SHEN
Chinese Journal of Tissue Engineering Research 2025;29(11):2357-2367
BACKGROUND:As a common disease in middle-aged and elderly,osteoarthritis is difficult to cure,and the pathogenesis is not clear.Long non-coding RNA participates in the pathogenesis of osteoarthritis through many ways,such as regulating translation,promoting or inhibiting mRNA,and adsorbing miRNAs. OBJECTIVE:To review the types of common long non-coding RNA in osteoarthritis,and the influence of multiple long non-coding RNAs on the pathological factors related to osteoarthritis,to analyze the future application of long non-coding RNAs in osteoarthritis. METHODS:Literature retrieval was conducted in CNKI,WanFang Data,VIP database,PubMed,Web of Science and Sciencedirect databases,using the search terms of"osteoarthritis,degenerative joint disease,degenerative arthritis,OA,LncRNA,long non-coding RNA,long noncoding RNA,long intergenic non-coding RNA"in Chinese and English.All relevant literature published from 1976 and May 2024 was retrieved.After literature screening,induction,analysis and summary,93 articles were finally included for review. RESULTS AND CONCLUSION:This review collected 25 long non-coding RNAs that are well studied with osteoarthritis.Long non-coding RNAs,as a molecular sponge for miRNA,are competing endogenous RNAs to competitively adsorb miRNAs and then affect downstream targets.Long non-coding RNAs can regulate physiopathological processes such as chondrocyte apoptosis and proliferation,cartilage extracellular matrix degradation,and inflammatory responses.Long non-coding RNAs are expected to become a biomarker and potential therapeutic target for the clinical diagnosis and therapeutic prognosis of osteoarthritis,and it may become a new strategy for the clinical treatment of osteoarthritis in the future.
7.Effect of Osteoking on Inflammatory Injury and NF-κB/NFATc1 Signaling Pathway in Model Mice with Postmenopausal Osteoporosis
Tiantian YU ; Ruibing FENG ; Qing WANG ; Chaowen LI ; Jiahao CHEN ; Ruixuan JIANG ; Yinshuai DING ; Yong HUANG ; Hao HU
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(10):62-69
ObjectiveTo explore the possible mechanism of Osteoking (OK) on postmenopausal osteoporosis (PMOP). MethodForty adult female mice were randomly divided into a sham operation (Sham) group, osteoporosis model (OVX) group, estradiol intervention (E2) group, and OK group, with 10 mice in each group. The modeling was completed by conventional back double incision ovariectomy, and the corresponding drugs were given one week later. After 12 weeks, the body mass and uterine index of mice were measured, and the pathological changes of bone tissue and the number of osteoclasts (OCs) were determined by hematoxylin-eosin (HE) and tartrate-resistant acid phosphatase (TRAP) staining, respectively. Bone mineral density (BMD), trabecular number (Tb.N), trabecular separation (Tb.Sp), and bone volume fraction (BV/TV) were measured by microcomputed tomography (Micro-CT). The maximum load of the femur was detected by a three-point bending test. The contents of tumor necrosis factor-α (TNF-α) and bone resorption marker C-terminal telopeptide of type Ⅰ collagen (CTX-1) were measured by enzyme linked immunosorbent assay (ELISA). The protein expression levels of nuclear factor-kappa B p65 (NF-κB p65), phosphorylated nuclear factor-kappa B p65 (p-NF-κB p65), nuclear factor kappa B inhibitor alpha (IκBα), phosphorylated nuclear factor kappa B alpha (p-IκBα), nuclear factor of activated T cells 1 (NFATc1), and proto-oncogene (c-Fos) were detected by Western blot. The mRNA expressions of OCs-related specific genes matrix metalloproteinase-9 (MMP-9), NFATc1, TRAP, cathepsin K (CTSK), and c-Fos were detected by real-time fluorescence quantitative polymerase chain reaction (Real-time PCR). ResultCompared with the Sham group, the uterine index decreased significantly in the OVX group, and the body mass (BMI) increased significantly. The structure of bone trabeculae was completely damaged, and the number of OCs increased. BMD, Tb.N, BV/TV, and maximum load decreased, while Tb.Sp was up-regulated. The levels of TNF-α and CTX-1 in serum were up-regulated. The protein expressions of c-Fos, p-NF-κB p65/NF-κB p65, NFATc1, and p-IκBα/IκBα were increased. The mRNA expressions of NFATc1, c-Fos, CTSK, TRAP, and MMP-9 were up-regulated (P<0.05, P<0.01). Compared with the OVX group, the body mass of the OK and E2 groups decreased, and the uterine index increased. The bone trabeculae increased, and the number of OCs decreased. BMD, Tb.N, BV/TV, and maximum load increased, while Tb.Sp decreased. The levels of TNF-α and CTX-1 in serum were decreased. The protein expressions of c-Fos, p-NF-κB p65/NF-κB p65, NFATc1, and p-IκBα/IκBα were decreased, and the mRNA expressions of NFATc1, c-Fos, CTSK, TRAP, and MMP-9 were decreased (P<0.05, P<0.01). ConclusionOK can inhibit the NF-κB/NFATc1 signaling pathway and reduce bone mass loss by reducing the level of inflammatory injury factors in PMOP mice, which is one of the mechanisms for treating PMOP.
8.Exploration on theoretical basis and approach of "Chinese materia medica transformation" of Indian natural medicine
International Journal of Traditional Chinese Medicine 2024;46(1):19-23
In history, there were frequent medical exchanges between China and foreign countries. After the introduction of foreign drugs, they are endowed with the properties of Chinese materia medica and undergo a process of utility evolution, which can be called "Chinese materia medica transformation". This article took natural drugs introduced from India as the research object and found that the common understanding between Ayurvedic medicine and TCM theory is the theoretical basis for the Chinese materia medica transformation" of natural drugs in India. Indian drugs need to complete the process of "Chinese materia medica transformation" through two approaches: the attribution of Chinese materia medica attributes based on the corresponding relationship between the two medical theories, and the supplementation of properties and effects beyond the common understanding. The case of "Chinese materia medica transformation" of natural drugs in India can provide a theoretical and practical basis for the research of "Chinese materia medica transformation" of more foreign new drugs.
9.Research Progress in Tong Du Tiao Shen of Mental Disorders
Jiahao ZHANG ; Cheng CHI ; Mengyue FAN ; Lin YAN ; Feixue WANG ; Meng ZHANG ; Yongjun CHEN
World Science and Technology-Modernization of Traditional Chinese Medicine 2024;26(1):61-68
Mental disorders are characterized by disturbances in behavior,volition,emotion,and cognition and are considered emotional diseases in traditional Chinese medicine.Acupuncture is one of the most widely used complementary alternative therapies for the treatment of mental disorders.Recently,there has been growing interest in the use of the Tong Du Tiao Shen(Dredging Du meridian to regulate the spirit)as a primary treatment.However,a comprehensive summary of the establishment and related acupuncture methods of Tong Du Tiao Shen is lacking.This paper aims to address this gap by exploring the origin and development of Tong Du Tiao Shen,its application in treating mental disorders,and the modern biological mechanisms involved.Ultimately,this paper seeks to expand the clinical application of Tong Du Tiao Shen acupuncture and provide a scientific basis for future research in this field.
10.The pathological progression of steroid-induced osteonecrosis of the femoral head caused by oxidative stress-induced osteoblast ferroptosis
Jiahao ZHANG ; Yuhao LIU ; Chi ZHOU ; Liang MO ; Hanjun FANG ; Zhenqiu CHEN
Chinese Journal of Tissue Engineering Research 2024;28(20):3202-3208
BACKGROUND:Studies have shown that imbalance of bone metabolism during glucocorticoid-induced osteonecrosis of the femoral head necrosis is closely related to oxidative stress. OBJECTIVE:To investigate the pathological mechanism by which oxidative stress-induced ferroptosis promote apoptosis in osteoblasts involved in steroid-induced osteonecrosis of the femoral head. METHODS:General data and serum specimens were collected from 47 patients with steroid-induced osteonecrosis of the femoral head.In addition,six femoral head specimens were collected from these patients.According to the Association Research Circulation Osseous(ARCO)staging system,serum specimens were grouped into ARCO Ⅱ,Ⅲ,and IV,while femoral head specimens were classified into ARCO Ⅲ and IV.Serum levels of malondialdehyde and superoxide dismutase 1 were measured.The protein expression of superoxide dismutase 1,glutathione peroxidase 4,Bcl-2 in the femoral head was detected and verified by Data independent acquisition(DIA)for quantitative sequencing,western blot and alkaline phosphate detection. RESULTS AND CONCLUSION:The ARCO stage of patients with steroid-induced osteonecrosis of the femoral head was independent of age,sex and necrotic side.The serum levels of malondialdehyde and superoxide dismutase 1 were higher in patients with ARCO stage Ⅲ compared with those with ARCO stage Ⅱ and IV.The results of DIA protein quantification showed that the function of differential proteins was mainly related to redox.The levels of superoxide dismutase 1,glutathione peroxidase 4,and Bcl-2 in the necrotic region were lower than in the normal region,as well as lower in ARCO stage IV than in ARCO stage Ⅲ.Western blot verified the results of DIA protein quantification.The alkaline phosphatase activity was lower in the necrotic region than in the normal region,as well as lower in ARCO stage IV than in ARCO stage Ⅲ.In the necrotic and sclerotic regions,the function of differential proteins was also related to redox,and superoxide dismutase 1,glutathione peroxidase 4,Bcl-2 protein expression and alkaline phosphatase activity were lower in the necrotic area than in the sclerotic region,as well as lower in ARCO stage IV than in ARCO stage Ⅲ.To conclude,glucocorticoids can influence the progression of steroid-induced osteonecrosis of the femoral head by upregulating oxidative stress levels,inducing osteoblast ferroptosis,and inhibiting osteogenic function.


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