1.Mechanistic Study on Tougu Xiaotong Capsules in Regulating PANoptosis to Delay Degeneration of Chondrocytes in Knee Osteoarthritis
Jinxia YE ; Yixin LIN ; Xiaoqing LEI ; Yanfeng HUANG ; Changlong FU ; Desen LI ; Wenyi WANG ; Lan WANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(8):149-161
ObjectiveTo investigate the effect of Tougu Xiaotong capsules (TGXTC) on the regulation of chondrocyte PANoptosis, delay of chondrocyte degeneration, and improvement of the symptoms in knee osteoarthritis (KOA). MethodsIn vivo experiments: 50 male C57BL/6 mice were randomly assigned into five groups (n=10 per group): sham operation group, model group, low-dose TGXTC group (7.2 g·kg-1), high-dose TGXTC group (14.4 g·kg-1), and diclofenac sodium group (0.05 g·kg-1). Except for the sham group, KOA models were established in all other groups using the modified Hulth method. Following successful model induction, the TGXTC groups received daily oral gavage of 7.2 or 14.4 g·kg-1 for 6 weeks, while the diclofenac sodium group received 0.05 g·kg-1 solution daily over the same duration. Model evaluation was performed using Lequesne MG score; micro-computed tomography (micro-CT) was used to scan the knee, hematoxylin-eosin (HE) staining and safranin O-fast green staining were used to observe the morphology of cartilage, transmission electron microscopy (TEM) was used to determine ultrastructural changes of PANoptosis. Multiple immunofluorescence (IF) co-localization assays was performed to detect the co-localization of cleaved Caspase-3, receptor-interacting protein 3 (RlPK3), and the N-terminal domain of gasdermin D (GSDMD-N) in cartilage tissue, while western blot was employed to detect the expression levels of cleaved Caspase-3, RIPK3, and GSDMD-N. In vitro experiments: The knee cartilages of 4-week-old SD rats were isolated, and a chondrocyte in vitro culture system was established through mechanical digestion with 0.2% type Ⅱ collagenase. Second-generation chondrocytes were divided into three groups: the control group, the model group (pretreated with 10 mg·L-1 lipopolysaccharide (LPS) for 24 h followed by treatment with 1 μmol·L-1 nigericin for 4 h), and the TGXTC treatment group (pretreated with 10 mg·L-1 LPS for 24 h, followed by exposure to 1 μmol·L-1 nigericin for 4 h and subsequently treated with 100 mg·L-1 TGXTC for an additional 24 h). The levels of reactive oxygen species (ROS), apoptosis, necroptosis, and pyroptosis of chondrocytes were evaluated via fluorescence microscopy following staining with ROS detection, AO/EB and YO-PRO-1/PI staining kits. Transmission electron microscopy was utilized to investigate the ultrastructural changes associated with PANoptosis in cartilage tissue of KOA mice. Inflammatory cytokine levels (IL-1β and IL-18) were measured using ELISA. Western blot was conducted to assess protein expressions related to PANoptosis, including cleaved Caspase-3, cleaved Caspase-8, RIPK3, ZBP1, GSDMD-N, and NLRP3. ResultsCompared with the sham group, the Lequesne MG scores were significantly up-regulated(P<0.01) in the model group, and the pathological changes of cartilage were significantly, with joint spaces narrower, osteophyte formation increased, secere abrasion of cartilage surface. Ultrastructural analysis revealed pronounced chondrocyte apoptosis, necroptosis, and pyroptosis, along with markedly elevated expression of cleaved Caspase-3, RlPK3, and GSDMD-N in cartilage tissue (P<0.01). In addition, The mean fluorescence intensities of ROS, orange-red fluorescence in AO/EB staining, green fluorescence and red fluorescence in YO-PRO-1/PI staining were increased of chondrocyte in the model group (P<0.01) . The levels of inflammatory factors IL-1β and IL-18 in the supernatant were increased (P<0.01). The expression of PANoptosis related proteins (cleaved Caspase-3, cleaved Caspase-8, RIPK3, ZBP1, GSDMD-N, and NLRP3) were also significantly upregulated(P<0.05). Compared to the model group, the TGXTC group demonstrated a significant improvement in various parameters of mice. These included a reduction in the Lequesne MG score, an increase in joint space, a decrease in osteophyte formation, diminished cartilage damage, reduced release of ROS, and alleviation of apoptotic, necroptotic, and pyroptotic processes in chondrocytes. Additionally, mitochondrial swelling and endoplasmic reticulum dilation were also mitigated. The levels of ROS as well as IL-1β and IL-18 were significantly decreased (P<0.05). Furthermore, the expression levels of proteins associated with PANoptosis in cartilage tissue showed marked reductions (P<0.05). Similar results were observed in chondrocytes: cleaved Caspase-3, cleaved Caspase-8, RIPK3, ZBP1, GSDMD-N, and NLRP3 exhibited significant decreases as well (P<0.05). ConclusionTGXTC may mitigate chondrocytes degeneration and alleviate KOA symptoms by reducing oxidative stress and suppressing the activation of PANoptosis pathways.
2.Mechanistic Study on Tougu Xiaotong Capsules in Regulating PANoptosis to Delay Degeneration of Chondrocytes in Knee Osteoarthritis
Jinxia YE ; Yixin LIN ; Xiaoqing LEI ; Yanfeng HUANG ; Changlong FU ; Desen LI ; Wenyi WANG ; Lan WANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(8):149-161
ObjectiveTo investigate the effect of Tougu Xiaotong capsules (TGXTC) on the regulation of chondrocyte PANoptosis, delay of chondrocyte degeneration, and improvement of the symptoms in knee osteoarthritis (KOA). MethodsIn vivo experiments: 50 male C57BL/6 mice were randomly assigned into five groups (n=10 per group): sham operation group, model group, low-dose TGXTC group (7.2 g·kg-1), high-dose TGXTC group (14.4 g·kg-1), and diclofenac sodium group (0.05 g·kg-1). Except for the sham group, KOA models were established in all other groups using the modified Hulth method. Following successful model induction, the TGXTC groups received daily oral gavage of 7.2 or 14.4 g·kg-1 for 6 weeks, while the diclofenac sodium group received 0.05 g·kg-1 solution daily over the same duration. Model evaluation was performed using Lequesne MG score; micro-computed tomography (micro-CT) was used to scan the knee, hematoxylin-eosin (HE) staining and safranin O-fast green staining were used to observe the morphology of cartilage, transmission electron microscopy (TEM) was used to determine ultrastructural changes of PANoptosis. Multiple immunofluorescence (IF) co-localization assays was performed to detect the co-localization of cleaved Caspase-3, receptor-interacting protein 3 (RlPK3), and the N-terminal domain of gasdermin D (GSDMD-N) in cartilage tissue, while western blot was employed to detect the expression levels of cleaved Caspase-3, RIPK3, and GSDMD-N. In vitro experiments: The knee cartilages of 4-week-old SD rats were isolated, and a chondrocyte in vitro culture system was established through mechanical digestion with 0.2% type Ⅱ collagenase. Second-generation chondrocytes were divided into three groups: the control group, the model group (pretreated with 10 mg·L-1 lipopolysaccharide (LPS) for 24 h followed by treatment with 1 μmol·L-1 nigericin for 4 h), and the TGXTC treatment group (pretreated with 10 mg·L-1 LPS for 24 h, followed by exposure to 1 μmol·L-1 nigericin for 4 h and subsequently treated with 100 mg·L-1 TGXTC for an additional 24 h). The levels of reactive oxygen species (ROS), apoptosis, necroptosis, and pyroptosis of chondrocytes were evaluated via fluorescence microscopy following staining with ROS detection, AO/EB and YO-PRO-1/PI staining kits. Transmission electron microscopy was utilized to investigate the ultrastructural changes associated with PANoptosis in cartilage tissue of KOA mice. Inflammatory cytokine levels (IL-1β and IL-18) were measured using ELISA. Western blot was conducted to assess protein expressions related to PANoptosis, including cleaved Caspase-3, cleaved Caspase-8, RIPK3, ZBP1, GSDMD-N, and NLRP3. ResultsCompared with the sham group, the Lequesne MG scores were significantly up-regulated(P<0.01) in the model group, and the pathological changes of cartilage were significantly, with joint spaces narrower, osteophyte formation increased, secere abrasion of cartilage surface. Ultrastructural analysis revealed pronounced chondrocyte apoptosis, necroptosis, and pyroptosis, along with markedly elevated expression of cleaved Caspase-3, RlPK3, and GSDMD-N in cartilage tissue (P<0.01). In addition, The mean fluorescence intensities of ROS, orange-red fluorescence in AO/EB staining, green fluorescence and red fluorescence in YO-PRO-1/PI staining were increased of chondrocyte in the model group (P<0.01) . The levels of inflammatory factors IL-1β and IL-18 in the supernatant were increased (P<0.01). The expression of PANoptosis related proteins (cleaved Caspase-3, cleaved Caspase-8, RIPK3, ZBP1, GSDMD-N, and NLRP3) were also significantly upregulated(P<0.05). Compared to the model group, the TGXTC group demonstrated a significant improvement in various parameters of mice. These included a reduction in the Lequesne MG score, an increase in joint space, a decrease in osteophyte formation, diminished cartilage damage, reduced release of ROS, and alleviation of apoptotic, necroptotic, and pyroptotic processes in chondrocytes. Additionally, mitochondrial swelling and endoplasmic reticulum dilation were also mitigated. The levels of ROS as well as IL-1β and IL-18 were significantly decreased (P<0.05). Furthermore, the expression levels of proteins associated with PANoptosis in cartilage tissue showed marked reductions (P<0.05). Similar results were observed in chondrocytes: cleaved Caspase-3, cleaved Caspase-8, RIPK3, ZBP1, GSDMD-N, and NLRP3 exhibited significant decreases as well (P<0.05). ConclusionTGXTC may mitigate chondrocytes degeneration and alleviate KOA symptoms by reducing oxidative stress and suppressing the activation of PANoptosis pathways.
3.Role of ABRACL in the occurrence and development of malignant tumors
Tongtong YE ; Zeyu WU ; Wenyi XI ; Zhiwei WANG ; Xiaochun JIANG ; Chenhui ZHAO
Journal of International Oncology 2023;50(9):544-547
The ABRACL protein, the regulator of actin and cell motility, belongs to the HSPC280 family, and its conserved hydrophobic groove can interact with other proteins to facilitate actin motility and cellular activity. ABRACL is upregulated in tumor tissues and is closely linked with the proliferation and migration of tumor cells. A deeper understanding of the role of ABRACL in tumorigenesis and development may provide new ideas and insights for ABRACL to prevent or reverse tumor progression.
4.Management and operation of extra-large Fangcang hospitals: experience and lessons from containing the highly contagious SARS-CoV-2 Omicron in Shanghai, China.
Yun XIAN ; Chenhao YU ; Minjie CHEN ; Lin ZHANG ; Xinyi ZHENG ; Shijian LI ; Erzhen CHEN ; Zhongwan CHEN ; Weihua CHEN ; Chaoying WANG ; Qingrong XU ; Tao HAN ; Weidong YE ; Wenyi XU ; Xu ZHUANG ; Yu ZHENG ; Min CHEN ; Jun QIN ; Yu FENG ; Shun WEI ; Yiling FAN ; Zhiruo ZHANG ; Junhua ZHENG
Frontiers of Medicine 2023;17(1):165-171
5.Microdissected peroneal artery perforator flap for repair soft tissue defect of dorsal fingers
Shaoxiao YU ; Wanggao ZHOU ; Guorong CHEN ; Zhenwei ZHANG ; Wenyi WU ; Xuelang YE ; Jinhao ZENG ; Yuhai KE
Chinese Journal of Microsurgery 2022;45(6):617-621
Objective:To investigate the clinical effect of microdissected peroneal artery perforator flap in repair of soft tissue defect of dorsal side of the fingers.Methods:From August 2015 to July 2020, 19 patients with soft tissue defects on dorsal fingers were treated with microdissected peroneal artery perforator flap. The area of wound defect was 3.8 cm×1.5 cm-5.8 cm×3.0 cm, with exposure of phalanges and tendons. The size of flaps was 4.0 cm×1.8 cm-6.0 cm×3.3 cm. According to the size of soft tissue defects on the dorsal side of the fingers, the flaps were designed with the perforating branch of peroneal artery in the centre. The length and width of a flap were 0.2-0.3 cm bigger and wider than the area of defect. The perforator vessels with a length of 2.0-3.0 cm were arvested in the superficial layer of deep fascia. Most of the adipose tissues of the flap were removed under microscope, and the small arteries between adipose tissues were protected. The flaps were used to cover the defects of fingers. The perforator artery of the flap was anastomosed with the proper palmar digital artery of the recipient site, the accompanying vein of the perforator artery was anastomosed with the dorsal digital vein of the recipient site, and the cutaneous nerve in the flap was anastomosed with the dorsal digital nerve. The donor sites were directly pulled together and sutured intermittently. Outpatient and WeChat follow-up were conducted after operation, including wound healing, flap survival, flap sensation, donor site recovery, and flexion and extension functions of the fingers. Functional recovery was evaluated according to the Evaluation Standard of Upper Limb Partial Functional of Hand Surgery of Chinese Medical Association.Results:All wounds healed in Ⅰ stage, and all 19 flaps survived. The follow-up ranged from 9 to 25 months, with an average of 11.5 months. The appearance of the flaps was satisfactory and the texture was good. Sensation recoveried to S 4 in 4 paitients, S 3 in 9 patients and S 2 in 6 patients, and with only a linear scar was left in the donor sites. The hand function recovery was evaluated according to the Trial Criteria of Upper Limb Function Evaluation of the Hand Surgery Society of the Chinese Medical Association, with 18 cases were excellent and 1 was good. Conclusion:The microdissected peroneal artery perforator flap is an ideal surgical method to repair the soft tissue defect of dorsal side of the fingers, which has good shape and simple operation, avoids the secondary thinning and plastic surgery and offers good therapeutic effects.
6.Research Process of Chemical Constituents and Pharmacological Effects of Tibetan Medicine Triphala
Kun ZHOU ; Ping JIAN ; Wenyi LIANG ; Linjin LIANG ; Yaping CUI ; Ting YE ; Lanzhen ZHANG
World Science and Technology-Modernization of Traditional Chinese Medicine 2018;20(9):1608-1614
Triphala consists of Terminalia chebula Retz, Terminalia bellirica (Gaertn.) Roxb. and Phyllanthus emblica L, which are made into Dasanguo Powder. Triphala is rich in a variety of chemical components, including tannins, phenolic acids, triterpenoids and flavonoids. The content of tannin is abundant in Triphala, which are often used as the main indicators of analysis. Modern research found that Triphala has a variety of pharmacological activities such as prevention of gastrointestinal diseases, antibacterial, anti-inflammatory, anti-oxidation, anti-tumor and so on. This paper briefly summarized the chemical constituents and pharmacological effects of Triphala, combining the relevant national and international literature in recent years to provide reference for the development and further study of Triphala.
7.Comprehensive Evaluation of the Quality of Tibetan Medicine Triphala's Tannin Parts Based on There Characteristic Components and the Total Tannin Content
Ting YE ; Yaping CUI ; Linjin LIANG ; Wenyi LIANG ; Ping JIAN ; Kun ZHOU ; LIngfang WU ; Shi LI ; Qi QI ; Lanzhen ZHANG
World Science and Technology-Modernization of Traditional Chinese Medicine 2018;20(9):1615-1621
To determine there characteristic components content of Tibetan Medicine Triphala through establishing a HPLC method and its total tannin content through spectrophotometry. The chromatographic column of Agilent ZORBAX SB-Aq (250 × 4.6 mm, 5 μm) with methanol-0.1% formic acid/water as the mobile phase, the flow rate 1 mL/min, and the detection wavelength 270 nm was applied to determine the Gallic acid, colijing and ellagic acid content in medicinal materials and the tannin parts of Tibetan Medicine Triphala. With the gallic acid as control group, total tannin content of Triphala and its tannin parts was determined through spectrophotometry. It revealed in the HPLC test that the linear range of gallic acid, coracine and ellagic acid was 0.91-4.55 μg, 0.274-1.368 μg and 0.329-2.634 μg respectively. It also showed that the average recovery rates of the three components in the medicinal materials were 101.06%, 101.72%and 100.27% respectively. And the average recovery rates of the three components in the tannins were 100.4%, 100.85%and 101.70% respectively. The result of spectrophotometry showed that gallic acid was linear in1.008-10.08 μg·mL-1, and that the recovery rate of medicinal materials and tannin parts were 100.25% and 100.52% respectively. The method is rapid, accurate and repeatable, and it can provide basis for the quality control of Tribescens and its tannins.
8.Study on Water-Soluble Constituents from Tibetan Medicine Terminalia billirica (Gaert.) Roxb
Shi LI ; Wenyi LIANG ; Linjin LIANG ; Ting YE ; Ping JIAN ; Kun ZHOU ; Lanzhen ZHANG
World Science and Technology-Modernization of Traditional Chinese Medicine 2018;20(9):1622-1626
This paper aimed at investigating the water-soluble constituents of Terminalia billirica (Gaert.) Roxb.Compounds were isolated and purified by Diaion HP-20, Sephadex LH-20, Toyopearl HW-40, MCI gel CHP 20 P and ODS column chromatographies. The chemical structures were elucidated according to spectral data and physicochemical properties. The results showed that ten compounds were isolated and identified as corilagin, salicoside, methyl-α-Darabinofuranoside, 1, 4-dimethyl-β-d-fructopyranose, β-fructopyranose, β-fructofuranose gallic acid, ellagic acid, 1, 2, 3, 4, 6-penta-O-galloyl-β-D-glucopyranonside, Chebulagic acid. Except compound 7 and 8, others compounds are separated from Terminalia billirica (Gaert.) Roxb for the first time, and except corilgin, the other compounds are separated from Terminalia for the first time.
9.Study on Rapid Identification of Constituents of Tibetan Medicine Terminalia chebular Retz. by HPLC/LTQ-Orbitrap MSn
Ping JIAN ; Qi QI ; Kun ZHOU ; Wenyi LIANG ; Linjin LIANG ; Ting YE ; Lanzhen ZHANG
World Science and Technology-Modernization of Traditional Chinese Medicine 2018;20(9):1627-1637
The aim of this work was to identify and analyze the chemical constituents of Terminalia chebular Retz by establishing high performance liquid chromatography-linear ion trap/electrostatic field orbitrap combined high resolution mass spectrometry (HPLC-LTQ-Orbitrap MS) technique. With the application of C18 reverse phase column (4.6 mm×250 mm, 5 μm), gradient elution was performed with methanol and water (0.2% acetic acid) as mobile phase.In negative ionization mode, the data of LTQ-Orbitrap was collected. Accurate molecular mass of molecular ion peaks and fragment ions provided by the high-resolution mass spectrometry were compared with the literature and the reference substance to determine the possible structure of the compound. The results indicated that 62 compounds from the extract of Terminalia chebular Retz. were identified, including acids and hydrolysable tannins (60), triterpenoids (2) . There were14 compounds were detected from this specie for the first time. It is proved as an effective method to provide reference for Chemical composition mass spectrometry and the quality control in further phytochemical studies of Terminalia chebular Retz.
10.Study on Quality Evaluation of Terminalia Chebular Based on Simultaneous Determination of Total Tannin and 7 Components
Linjin LIANG ; Qi QI ; Ting YE ; Wenyi LIANG ; Ping JIAN ; Kun ZHOU ; Lanzhen ZHANG
World Science and Technology-Modernization of Traditional Chinese Medicine 2018;20(9):1638-1644
The spectrophotometric method was established for the determination of total tannin content in gardenia medicinal materials and tannins, and HPLC method was applied to simultaneously determine gallic acid, punical glucoside A, methyl gallate, punny glucoside B, corilagin, pentagalloyl glucose and the content of ellagic acid. Taking gallic acid as a reference substance, a phosphomolybdate tungstic acid colorimetric method (Pharmacopoeia method) was used to determine the total tannin content. With the chromatographic column Agilent ZORBAX SB-Aq-C18 (250 × 4.6 mm, 5 microns), mobile phase for phosphoric acid (0.1%), methanol, water flow rate of 1 ml/min, column temperature 30℃, detection wavelength of 270 nm, gradient elution, 7 kinds of component contents in myrobalan medicinal materials were measured at the same time. The result showed that the average content of total tannins of Radix Scutellariae was 30.31%, RSD was 0.55%; the average recovery rate was 99.70%, and the RSD was 21.97% (n = 6) . In the scorpion medicinal materials, gallic acid, punical glucoside A, punny glucoside B, methyl gallate, punny glucoside B, creatin, pentagalloglucose and ellagic acid have a good linear relationship in their respective ranges. The recovery rate of the sample is between100.10% and 102.77%. The establishment of the determination method of total tannin, gallic acid, punical glucoside, methyl gallate, corridain, pentagalloglucose and ellagic acid in the scorpion medicinal materials is simple and accurate, which has strong specificity and can be used for quality control of Terminalia chebular.

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