1.Allogeneic lung transplantation in miniature pigs and postoperative monitoring
Yaobo ZHAO ; Ullah SALMAN ; Kaiyan BAO ; Hua KUI ; Taiyun WEI ; Hongfang ZHAO ; Xiaoting TAO ; Xinzhong NING ; Yong LIU ; Guimei ZHANG ; He XIAO ; Jiaoxiang WANG ; Chang YANG ; Feiyan ZHU ; Kaixiang XU ; Kun QIAO ; Hongjiang WEI
Organ Transplantation 2026;17(1):95-105
Objective To explore the feasibility and reference value of allogeneic lung transplantation and postoperative monitoring in miniature pigs for lung transplantation research. Methods Two miniature pigs (R1 and R2) underwent left lung allogeneic transplantation. Complement-dependent cytotoxicity tests and blood cross-matching were performed before surgery. The main operative times and partial pressure of arterial oxygen (PaO2) after opening the pulmonary artery were recorded during surgery. Postoperatively, routine blood tests, biochemical blood indicators and inflammatory factors were detected, and pathological examinations of multiple organs were conducted. Results The complement-dependent cytotoxicity test showed that the survival rate of lymphocytes between donors and recipients was 42.5%-47.3%, and no agglutination reaction occurred in the cross-matching. The first warm ischemia times of D1 and D2 were 17 min and 10 min, respectively, and the cold ischemia times were 246 min and 216 min, respectively. Ultimately, R1 and R2 survived for 1.5 h and 104 h, respectively. Postoperatively, in R1, albumin (ALB) and globulin (GLB) decreased, and alanine aminotransferase increased; in R2, ALB, GLB and aspartate aminotransferase all increased. Urea nitrogen and serum creatinine increased in both recipients. Pathological results showed that in R1, the transplanted lung had partial consolidation with inflammatory cell infiltration, and multiple organs were congested and damaged. In R2, the transplanted lung had severe necrosis with fibrosis, and multiple organs had mild to moderate damage. The expression levels of interleukin-1β and interleukin-6 increased in the transplanted lungs. Conclusions The allogeneic lung transplantation model in miniature pigs may systematically evaluate immunological compatibility, intraoperative function and postoperative organ damage. The data obtained may provide technical references for subsequent lung transplantation research.
2.Reflections on Status Quo and Development Pathways of Traditional Chinese Medicine Technology Transfer in Context of Digital-intelligent Transformation
Jie ZHANG ; Jing XU ; Guangwei ZHENG ; Huayu ZHANG ; Chang LIU ; Xiaoxiao WEN ; Xishui PAN ; Bin WANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(4):235-240
As a distinctive resource of Chinese civilization, traditional Chinese medicine (TCM) technology transfer faces significant opportunities under the background of digital and intelligent transformation, while also being constrained by unique challenges such as the complexity of its theoretical system, lengthy industrial chains, and multidimensional policy restrictions, resulting in a "high-value-high-threshold" paradox. At present, TCM technology transfer is deeply trapped in a "threefold reluctance" dilemma, i.e., unwillingness to transfer, inability to transfer, and lack of capacity to transfer. Specifically, the disconnection between scientific research evaluation systems and market demand leads to low conversion rates of research achievements, unclear ownership and compliance risks suppress innovation incentives, and the absence of professional services intensifies supply-demand mismatches. This article systematically analyzes the specific characteristics of TCM technology transfer and proposes a breakthrough pathway centered on full-chain digital and intelligent transformation. By integrating technologies such as intelligent sorting systems, blockchain-based traceability, and AI diagnostic models, the TCM ecosystem spanning "cultivation-production-service" can be reconstructed. In terms of standardization, promoting the progression from "experience-based data conversion" to "data standardization" and further to "intelligent standardization" is advocated to resolve quality control challenges. For example, a "three-no-one-full" certification system can strengthen quality trust. Policy coordination should focus on optimizing mechanisms for the transformation of scientific and technological achievements, while exploring intellectual property securitization and risk-sharing models to stimulate research momentum. In terms of internationalization, reliance on the Belt and Road Initiative platform to promote the export of geo-authentic medicinal material brands and standards is recommended to build a dual-driven model of "technology plus culture". Looking ahead, through the construction of national-level databases, the cultivation of interdisciplinary talent, and the mutual recognition of international standards, a new paradigm of "scientific intelligent manufacturing" can be formed, providing systematic solutions for the modernization of TCM and global health governance.
3.Two cases of acute radiation-induced skin injury caused by external exposure to 192Ir
Li LI ; Wei SHANG ; Yan LING ; Mi WANG ; Huisheng ZHANG ; Chiqiao LU ; Xiaohu ZHONG ; Shenglong XU ; Juan GUO ; Chang LIU ; Yulong LIU
Chinese Journal of Radiological Health 2026;35(1):56-61
Objective To introduce the causes of accidents and the diagnosis and treatment of two patients with radiation-induced skin injury admitted to our hospital in 2023, and to provide a reference for the clinical treatment of subsequent radiation-induced skin injury. Methods The clinical treatment process of two patients with acute skin injury caused by external radiation exposure were summarized and analyzed. Results The exposure history of the two patients was reconstructed, the flaw detection scenario was simulated, the biological dose and hand skin exposure dose were estimated, and the infrared thermal imaging device was used for dynamic monitoring. A comprehensive analysis was conducted based on clinical manifestations and other data. The diagnosis of “Xie” was excessive exposure combined with acute radiation-induced skin injury on both hands (Grade IV for the right hand palm, index finger, and middle finger and Grade II for the left hand little finger). The diagnosis of “Hao” was acute radiation-induced skin injury on both hands (Grade I). The two patients received different clinical treatment measures: “Xie” was treated with both local and systemic therapies, while “Hao” was mainly treated with systemic therapy. Conclusion After systematic and effective treatment, the radiation-induced skin injuries healed in both patients.
4.Flavonoids Intervene in Diabetic Nephropathy by Regulating TGF-β/Smad Signaling Pathway: A Review
Qihui QIU ; Chang LIU ; Xiaotong YAN ; Jinwei HAN ; Hui SUN ; Fengting YIN ; Yuhang WANG ; Mengmeng WANG ; Xijun WANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(7):300-309
Diabetic nephropathy (DKD), as a common microvascular complication of diabetes mellitus (DM), is a major cause of end-stage renal disease (ESRD). Its clinical manifestations include increased urinary protein excretion, thickening of the glomerular basement membrane, and renal tubulointerstitial fibrosis. The pathogenesis of DKD is complex and involves multiple factors, including disordered glucose metabolism, hemodynamic alterations, and oxidative stress. Although modern medical approaches can alleviate certain symptoms, they still have limitations such as insufficient therapeutic targeting and prominent adverse effects. The transforming growth factor-β/Smad (TGF-β/Smad) signaling pathway is not only a tissue fibrosis pathway that has attracted considerable attention in recent years, but also regulates multiple protein molecules, including the glomerular podocyte slit diaphragm protein Podocin, interleukin-1β (IL-1β), and superoxide dismutase (SOD), thereby participating in various pathological processes and ultimately mediating renal injury. Flavonoid compounds, owing to their sustained pharmacological effects, broad spectrum of action, and high safety profile, have become ideal candidates for targeted therapy research in DKD. Existing studies have shown that these compounds can exert inhibitory effects on renal fibrosis, alleviate inflammatory responses, protect podocytes, and reduce oxidative stress by regulating the interactions between the TGF-β/Smad signaling pathway and the aforementioned protein molecules, thereby maintaining renal structure and function, reducing proteinuria, and significantly improving DKD lesions. This review briefly outlines the composition and functions of the TGF-β/Smad signaling pathway, elucidates the mechanisms by which this pathway regulates DKD, and focuses on summarizing major studies from the past decade on flavonoid-based interventions in DKD through targeted inhibition of the TGF-β/Smad signaling pathway. Furthermore, it discusses the considerable therapeutic potential of flavonoids in the treatment of this disease, aiming to provide a scientific basis for future clinical prevention and treatment of DKD and to promote the development of targeted drugs.
5.Impact of number of positive regional lymph nodes in N1 stage on the prognosis of patients with non-small cell lung cancer: A propensity score matching study
Dandan LIU ; Jiachen WANG ; Lidan CHANG ; Jia CHEN ; Ranran KONG ; Shiyuan LIU ; Minxia ZHU ; Jiantao JIANG ; Shaomin LI ; Zhengshui XU
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2026;33(01):63-71
Objective To explore the impact of number of positive regional lymph nodes (nPRLN) in N1 stage on the prognosis of non-small cell lung cancer (NSCLC) patients. Methods Patients with TxN1M0 stage NSCLC who underwent lobectomy and mediastinal lymph node dissection from 2010 to 2015 were screened from SEER database (17 Regs, 2022nov sub). The optimal cutoff value of nPRLN was determined using X-tile software, and patients were divided into 2 groups according to the cutoff value: a nPRLN≤optimal cutoff group and a nPRLN>optimal cutoff group. The influence of confounding factors was minimized by propensity score matching (PSM) at a ratio of 1 : 1. Kaplan-Meier curves and Cox proportional hazards models were used to evaluate overall survival (OS) and lung cancer-specific survival (LCSS) of patients. Results A total of 1316 patients with TxN1M0 stage NSCLC were included, including 662 males and 654 females, with a median age of 67 (60, 73) years. The optimal cutoff value of nPRLN was 3, with 1165 patients in the nPRLN≤3 group and 151 patients in the nPRLN>3 group. After PSM, there were 138 patients in each group. Regardless of before or after PSM, OS and LCSS of patients in the nPRLN≤3 group were superior to those in the nPRLN>3 group (P<0.001). N1 stage nPRLN>3 was an independent prognostic risk factor for OS [HR=1.52, 95%CI (1.22, 1.89), P<0.001] and LCSS [HR=1.72, 95%CI (1.36, 2.18), P<0.001]. Conclusion N1 stage nPRLN>3 is an independent prognostic risk factor for NSCLC patients in TxN1M0 stage, which may provide new evidence for future revision of TNM staging N1 stage subclassification.
6.Study on Reducing Hepatotoxicity and Retaining Anti-osteoporosis Activity of Psoraleae Fructus Though Salt Processing Based on Zebrafish
Yiqi LIU ; Xuan WANG ; Qiqi FAN ; Zehua CHANG ; Shuo FAN ; Na WANG ; Zheng LI ; Xinfang XU ; Chongjun ZHAO ; Xiangri LI
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(9):79-90
ObjectiveTo investigate the mechanism of salt processing of Psoraleae Fructus (PF) through modern analytical techniques and biotechnology, focusing on its effects related to hepatotoxicity and anti-osteoporosis activity. MethodsThe zebrafish model was utilized to evaluate the impact of PF and salt-processed Psoraleae Fructus (SPF) on the hepatotoxicity (using 134.17 , 178.89, 268.34 mg·L-1 as low, medium, and high dose groups of PF, 135.04, 180.06, 270.08 mg·L-1 as low, medium, and high dose groups of SPF, respectively) and anti-osteoporotic activity (using 33.54 , 67.08 and 134.17 mg·L-1 as low, medium, and high dose groups of PF, 33.76, 67.52, 135.04 mg·L-1 as low, medium, and high dose groups of SPF, respectively), which was using alizarin red skull staining of zebrafish as an indicator of different batches of PF. The specific dosage of a batch of PF was taken as an example. Then ultra-performance liquid chromatography-quadrupole-time of flight-mass spectrometry(UPLC-Q-TOF-MS) analysis was employed to identify the chemical composition of PF before and after salt processing, and PCA, OPLS-DA, and independent sample t-test were used to elucidating the compositional changes associated with the effects of salt processing on hepatotoxicity and anti-osteoporosis activity. ResultsUnder specific conditions, PF induced notable hepatotoxicity in zebrafish while simultaneously demonstrating protective effect against prednisolone-induced osteoporosis. In comparison to PF, SPF showed alleviated hepatotoxicity while retaining significant anti-osteoporosis activity. UPLC-Q-TOF-MS analysis revealed that after salt processing, the overall chemical composition of PF showed a downward trend, with 69 components showing a decrease in content, represented by psoralen, and 13 components showing an increase, represented by 4′-O-methyl psoralen B. Further multivariate statistical analysis revealed 11 key differential components before and after salt processing of PF, including psoralen and bakuchiol. ConclusionSalt processing effectively diminishes hepatotoxicity without impairing therapeutic efficacy against osteoporosis of PF, which may be related to the compositional changes before and after salt processing of PF and provides key evidence to reveal the scientific significance of salt processing of PF.
7.Mechanism of Bushen Kaixuan Tongluo Prescription in Improving Diabetic Nephropathy Based on cAMP Signaling Pathway
Miao XU ; Baosheng ZHAO ; You WANG ; Yuzhuo CHANG ; Zehao LIU ; Lingling QIN ; Haiyan WANG ; Ming GAO ; Cuiyan LYU ; Tonghua LIU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(11):87-96
ObjectiveTo investigate the molecular mechanism by which the Bushen Kaixuan Tongluo prescription exerts a renal protective effect in mice with diabetic kidney disease (DKD) by regulating the cyclic adenosine monophosphate (cAMP) signaling pathway. MethodsThirty specific pathogen-free (SPF) male db/db mice were adaptively fed for three weeks. Mice with a random tail vein blood glucose level ≥ 11.1 mmol·L-1 and urinary albumin-creatinine ratio (ACR) ≥ 30 mg·g-1 were considered successfully modeled. The successfully modeled mice were randomly divided into five groups with six mice in each group: the model group, the low-, medium-, and high-dose Bushen Kaixuan Tongluo prescription groups (administered at doses of 7, 14, 28 g·kg-1·d-1 respectively), and the positive drug irbesartan group (administered at a dose of 20 mg·kg-1·d-1). Additionally, six db/m mice were selected as the blank group. Mice in each group were given intragastric administration of the Bushen Kaixuan Tongluo prescription at the corresponding concentrations, irbesartan, or an equal volume of pure water, and the intervention lasted for 12 weeks. During the experiment, the general conditions, body weight changes, and renal function indicators of the mice were dynamically monitored. After the intervention, a blood glucose meter was used to measure the fasting blood glucose (FBG) of the mice. An automatic biochemical analyzer was employed to detect the levels of serum creatinine (SCr), blood urea nitrogen (BUN), urinary microalbumin (uALB), ACR, aspartate aminotransferase (AST), alanine aminotransferase (ALT), total cholesterol (TC), triglycerides (TG), leptin (LEP), glycosylated serum protein (GSP), and insulin (INS) in the mice. Renal tissues were collected for hematoxylin-eosin (HE) staining, periodic acid-Schiff (PAS) staining, and Masson's trichrome staining to observe the histopathological changes. Immunohistochemistry (IHC) was used to detect the expressions of protein kinase A (PKA) and cAMP response element-binding protein (CREB) in the mice. Western blot analysis was performed to determine the expression levels of PKA, phosphorylated protein kinase A (p-PKA), CREB, phosphorylated cAMP response element-binding protein (p-CREB), and B-cell lymphoma-2 (Bcl-2) proteins in the renal tissues of the mice. Real-time quantitative polymerase chain reaction (Real-time PCR) was used to detect the mRNA expression levels of PKA, CREB, and Bcl-2 in the renal tissues of the mice. ResultsCompared with the blank group, the mice in the model group showed listlessness, decreased activity, and a significant increase in body weight (P<0.01). Biochemical indicators revealed that the levels of BUN, uALB, ACR, AST, ALT, TC, TG, FBG, LEP, GSP, and INS were significantly increased (P<0.01), while SCr showed an increasing trend with no statistically significant difference. Compared with the model group, the mice in the Bushen Kaixuan Tongluo prescription intervention groups had improved general conditions and a decreasing trend in body weight. Biochemical indicators showed that the levels of BUN, uALB, ACR, TC, GSP, and INS were significantly decreased (P<0.05), while SCr, AST, ALT, TG, and LEP showed a decreasing trend with no statistically significant difference. Renal histopathological analysis showed that the model group exhibited typical DKD pathological features such as thickening of the glomerular basement membrane, expansion of the mesangial matrix, and deposition of collagen fibers in the renal tubulointerstitium, and all treatment groups could alleviate the above pathological damages. The IHC results showed that compared with the blank group, the expression levels of p-PKA and p-CREB in the renal tissues of the model group were significantly decreased (P<0.01). Compared with the model group, the expression level of p-PKA in the medium-dose Bushen Kaixuan Tongluo prescription group was significantly increased (P<0.01), while the expression level of p-CREB showed an increasing trend with no statistically significant difference. Western blot results showed that compared with the blank group, the expression levels of p-PKA/PKA, p-CREB/CREB, and Bcl-2 in the model group were significantly decreased (P<0.05). Compared with the model group, the expression levels of these proteins in the medium-dose Bushen Kaixuan Tongluo prescription group were significantly increased (P<0.01). Real-time PCR results showed that compared with the blank group, the mRNA expressions of PKA, CREB, and Bcl-2 in the model group were significantly down-regulated (P<0.05). Compared with the model group, the mRNA expressions of these genes in the medium-dose Bushen Kaixuan Tongluo prescription group were significantly up-regulated (P<0.05). ConclusionThe Bushen Kaixuan Tongluo prescription can improve the liver and kidney functions of db/db mice, correct lipid metabolism disorders and glucose metabolism imbalance. Its renal protective effect is associated with up-regulating the cAMP signaling pathway to improve renal fibrosis and reduce the level of oxidative stress, thereby protecting renal function.
8.Erchentang Ameliorates SiO2-induced Lung Injury by Regulating Oxidative Stress and Metabolic Disorders via Nrf2/HO-1 Signaling Pathway
Jun LU ; Xinyi ZHU ; Ziyi LIU ; Jixia HU ; Jialu CHEN ; Rong XIAO ; Zhibin WANG ; Chang LIU ; Fangguo LU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(13):32-42
ObjectiveTo observe the protective effect of Erchentang (ECT) on SiO2-induced lung injury in rats and to explore its underlying mechanism. MethodsA rat model of lung injury was established by a single intratracheal instillation of 50 mg·mL-1 SiO2 suspension. Thirty male Sprague-Dawley (SD) rats were randomly assigned to five groups: control, model, low and high-dose (4.5 g·kg-1·d-1 and 9 g·kg-1·d-1, respectively) ECT, and dexamethasone (0.2 mg·kg-1·d-1). All the groups were treated for 4 consecutive weeks. Histopathological alterations in the lung tissue were examined by hematoxylin and eosin (HE) staining. The levels of malondialdehyde (MDA), superoxide dismutase (SOD), and glutathione peroxidase (GSH-Px) in the lung tissue were measured through biochemical assays. The expression of key molecules in the nuclear factor erythroid 2-related factor 2 (Nrf2)/heme oxygenase-1 (HO-1) pathway was determined by Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR), Western blot, and immunofluorescence assay. The primary active components of ECT were identified by ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS), and their binding affinity to Nrf2/HO-1 was assessed by molecular docking. Untargeted metabolomics of the lung tissue was performed based on UPLC-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS), and correlation analysis was performed to identify differential metabolites and parameters closely associated with the Nrf2/HO-1 pathway. ResultsCompared with the control group, the model group exhibited a reduction in body weight gain, an increase in lung index, increased MDA content, weakened SOD and GSH-Px activities in the lung tissue, down-regulated mRNA and protein levels of Nrf2 and protein levels of HO-1 and GPX4, and an up-regulated protein level of Keap1 (P<0.05, P<0.01). Treatment with ECT attenuated the SiO2-induced decline in body weight (P<0.05), alleviated inflammatory cell infiltration and silicotic nodule formation in alveoli, and reduced the MDA content and enhanced the SOD and GSH-Px activities in the lung tissue (P<0.05, P<0.01). UPLC-MS/MS and molecular docking revealed that core components of ECT, such as hesperidin and glycyrrhizic acid, displayed strong binding affinity to Nrf2/HO-1. Molecular biological experiments demonstrated that ECT promoted nuclear translocation of Nrf2, up-regulated the mRNA and protein levels of HO-1 and GPX4, and down-regulated Keap1 expression (P<0.05, P<0.01). Metabolomic analysis indicated that ECT reversed the SiO2-induced aberrant expression of metabolites, including linoleic acid and glutamine (P<0.05, P<0.01). Correlation analysis showed that Nrf2 and HO-1 were positively correlated with SOD and GSH-Px (P<0.05, P<0.01), but negatively correlated with glutamine and serine (P<0.05, P<0.01). ConclusionECT may activate the Nrf2/HO-1 pathway through its core active components, thereby regulating oxidative stress and metabolic disorders to ameliorate SiO2-induced lung injury in rats. This study provides experimental evidence for ECT in the prevention and treatment of occupational lung injury.
9.Material Basis and Its Distribution in vivo of Qili Qiangxin Capsules Analyzed by UPLC-Q-Orbitrap-MS
Jianwei ZHANG ; Jiekai HUA ; Rongsheng LI ; Qin WANG ; Xinnan CHANG ; Wei LIU ; Jie SHEN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(5):185-193
ObjectiveBased on ultra-performance liquid chromatography-quadrupole-electrostatic field orbitrap high resolution mass spectrometry(UPLC-Q-Orbitrap-MS), the chemical constituents of Qili Qiangxin capsules was identified, and their distribution in vivo was analyzed. MethodsUPLC-Q-Orbitrap-MS was used to detect the sample solution of Qili Qiangxin capsules, as well as the serum, brain, heart, lung, spleen, liver and kidney tissues of mice after oral administration. Using the Thermo Xcalibur 2.2 software, the compound information database was constructed, and the molecular formulas of compounds corresponding to the quasi-molecular ions were fitted. Based on the information of retention time, accurate relative molecular mass and fragments, the compounds and their distribution in vivo were analyzed by comparing with the data of reference substances and literature. ResultsA total of 233 compounds, including 70 terpenoids, 60 flavonoids, 23 organic acids, 17 alkaloids, 20 steroids, 7 coumarins and 36 others, were identified or predicted from Qili Qiangxin capsules, 73 of which were identified matching with standard substances. Tissue distribution results showed that 71, 17, 38, 33, 32, 58 and 43 migrating components were detected in blood, brain, heart, lung, spleen, liver and kidney, respectively. Thirty-seven components were absorbed into the blood and heart, including quinic acid, benzoylaconitine benzoylmesaconine and so on. Fourteen components were absorbed into the blood and six tissues, including calycosin, methylnissolin, formononetin, alisol B, alisol A and so on. ConclusionThis study comprehensively analyzes the chemical components of Qili Qiangxin capsules and their distribution in vivo. Among them, astragaloside Ⅳ, salvianolic acid B, ginsenoside Rb1, ginsenoside Rb3, ginsenoside Rd, ginsenoside Rg3, calycosin-7-glucoside, and sinapine may be the important components for the treatment of heart failure, which can provide useful reference for its quality control and research on pharmacodynamic material basis.
10.Molecular biological research and molecular homologous modeling of Bw.03 subgroup
Li WANG ; Yongkui KONG ; Huifang JIN ; Xin LIU ; Ying XIE ; Xue LIU ; Yanli CHANG ; Yafang WANG ; Shumiao YANG ; Di ZHU ; Qiankun YANG
Chinese Journal of Blood Transfusion 2025;38(1):112-115
[Objective] To study the molecular biological mechanism for a case of ABO blood group B subtype, and perform three-dimensional modeling of the mutant enzyme. [Methods] The ABO phenotype was identified by the tube method and microcolumn gel method; the ABO gene of the proband was detected by sequence-specific primer polymerase chain reaction (PCR-SSP), and the exon 6 and 7 of the ABO gene were sequenced and analyzed. Homologous modeling of Bw.03 glycosyltransferase (GT) was carried out by Modeller and analyzed by PyMOL2.5.0 software. [Results] The weakening B antigen was detected in the proband sample by forward typing, and anti-B antibody was detected by reverse typing. PCR-SSP detection showed B, O gene, and the sequencing results showed c.721 C>T mutation in exon 7 of the B gene, resulting in p. Arg 241 Trp. Compared with the wild type, the structure of Bw.03GT was partially changed, and the intermolecular force analysis showed that the original three hydrogen bonds at 241 position disappeared. [Conclusion] Blood group molecular biology examination is helpful for the accurate identification of ambiguous blood group. Homologous modeling more intuitively shows the key site for the weakening of Bw.03 GT activity. The intermolecular force analysis can explain the root cause of enzyme activity weakening.

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