1.Effect of Yiqi Wenyang Huoxue Lishui Components on Cardiac Function and Mitochondrial Energy Metabolism in CHF Rats
Hui GAO ; Zeqi YANG ; Xin LIU ; Fan GAO ; Yangyang HAN ; Aiyangzi LU ; Xingchao LIU ; Qiuhong GUO
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(2):27-36
ObjectiveTo investigate the effects of Yiqi Wenyang Huoxue Lishui components on the cardiac function and mitochondrial energy metabolism in the rat model of chronic heart failure (CHF) and explore the underlying mechanism. MethodsThe rat model of CHF was prepared by transverse aortic constriction (TAC). Eight of the 50 SD rats were randomly selected as the sham group, and the remaining 42 underwent TAC surgery. The 24 SD rats successfully modeled were randomized into model, trimetazidine (6.3 mg·kg-1), and Yiqi Wenyang Huoxue Lishui components (60 mg·kg-1 total saponins of Astragali Radix, 10 mg·kg-1 total phenolic acids of Salviae Miltiorrhizae Radix et Rhizoma, 190 mg·kg-1 aqueous extract of Lepidii Semen, and 100 mg·kg-1 cinnamaldehyde) groups. The rats were administrated with corresponding agents by gavage, and those in the sham and model groups were administrated with the same amount of normal saline at a dose of 10 mL·kg-1 for 8 weeks. Echocardiography was used to examine the cardiac function in rats. Enzyme-linked immunosorbent assay was employed to determine the serum levels of N-terminal pro-B-type natriuretic peptide (NT-ProBNP), hypersensitive troponin(cTnI), creatine kinase (CK), lactate dehydrogenase (LD), free fatty acids (FFA), superoxide dismutase (SOD), and malondialdehyde (MDA). The colorimetric assay was employed to measure the levels of adenosine triphosphate (ATP), adenosine diphosphate (ADP), and adenosine monophosphate (AMP) in the myocardial tissue. The pathological changes in the myocardial tissue were observed by hematoxylin-eosin staining and Masson staining. The Na+-K+-ATPase and Ca2+-Mg2+-ATPase activities in the myocardial tissue were determined by the colorimetric assay. The ultrastructural changes of myocardial mitochondria were observed by transmission electron microscopy. Western blot was employed to determine the protein levels of ATP synthase subunit delta (ATP5D), glucose transporter 4 (GLUT4), and carnitine palmitoyltransferase-1 (CPT-1). The mitochondrial complex assay kits were used to determine the activities of mitochondrial complexes Ⅰ, Ⅱ, Ⅲ, and Ⅳ. ResultsCompared with the sham group, the model group showed a loosening arrangement of cardiac fibers, fracture and necrosis of partial cardiac fibers, inflammatory cells in necrotic areas, massive blue fibrotic tissue in the myocardial interstitium, increased collagen fiber area and myocardial fibrosis, destroyed mitochondria, myofibril disarrangement, sparse myofilaments, and fractured and reduced cristae. In addition, the rats in the model group showed declined ejection fraction (EF) and fractional shortening (FS), risen left ventricular end-diastolic diameter (LVIDd), left ventricular end-systolic diameter (LVIDs), left ventricular end-diastolic posterior wall thickness (LVPWd), left ventricular end-systolic posterior wall thickness (LVPWs), left ventricular end-diastolic volume (LVVOLd), and left ventricular end-systolic volume (LVVOLs), elevated levels of NT-ProBNP, cTnI, CK, MDA, FFA, and LD, lowered level of SOD, down-regulated protein levels of GLUT4 and CPT-1, decreased activities of Na+-K+-ATPase, Ca2+-Mg2+-ATPase, and respiratory complexes Ⅰ-Ⅳ, and declined levels of ATP5D, ATP, ADP, and AMP (P<0.05, P<0.01). Compared with the model group, the Yiqi Wenyang Huoxue Lishui components and trimetazidine groups showed alleviated pathological damage of the mitochondria and mycardial tissue, risen EF and FS, declined LVIDd, LVIDs, LVPWd, LVPWs, LVVOLd, and LVVOLs, lowered levels of NT-ProBNP, cTnI, CK, MDA, FFA, and LD, elevated level of SOD, up-regulated protein levels of GLUT4 and CPT-1, increased activities of Na+-K+-ATPase, Ca2+-Mg2+-ATPase, and respiratory complexes Ⅰ-Ⅳ, and elevated levels of ATP5D, ATP, ADP, and AMP (P<0.05, P<0.01). ConclusionYiqi Wenyang Huoxue Lishui components can improve the cardiac function, reduce myocardial injury, regulate glucose and lipid metabolism, optimize the utilization of substrates, and alleviate the damage of mitochondrial structure and function, thus improving the energy metabolism of the myocardium in the rat model of CHF.
2.Effects and mechanism of astilbin on renal injury in chronic renal failure rats
Xiaowei GAO ; Yingying LIU ; Cong HAN ; Shifei HAO
China Pharmacy 2025;36(4):434-439
OBJECTIVE To investigate the effect and potential mechanism of astilbin (AST) on renal injury in chronic renal failure (CRF) rats based on the Jagged-1/Notch-1 signaling pathway. METHODS CRF model was constructed by 5/6 nephrotomy. The successfully modeled rats were randomly separated into Model group, AST low-dose group (AST-L group), AST high-dose group (AST-H group), high-dose of AST+Notch pathway activator (Jagged-1/FC chimerin, referred to as “JFC”) group (AST-H+ JFC group), and control group (CK group) for open surgery without resection was set up, with 10 rats in each group. The rats in the AST-L group and AST-H group were given 40 and 80 mg/kg AST, respectively; the rats in the AST-H+JFC group were simultaneously given 80 mg/kg AST and 0.5 mg/kg JFC, and the rats in the CK group and Model group were given an equal volume of normal saline, once a day, for 4 weeks. After the last administration, the serum levels of blood urea nitrogen (BUN), serum creatinine (SCr), and the level of 24 h urinary protein (UP) in urine, as well as the serum levels of lactate dehydrogenase (LDH), tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and IL-10 in each group were detected. The morphology and fibrosis of renal tissue were observed. The content of adenosine triphosphate (ATP) and the activities of sodium-potassium ATPase and calcium-magnesium ATPase in mitochondria of renal tissue were detected. The protein expressions of transforming growth factor- β (TGF- β), hypoxia-inducible factor-1α (HIF-1α), α-smooth muscle actin (α-SMA), cleaved-caspase-3, Jagged-1 and Notch-1 in renal tissue were also observed. RESULTS Compared with CK group, the renal tissue of rats in the Model group was obviously damaged, renal tissue fibrosis was severe; the serum BUN and SCr levels, urine UP level, serum 4 LDH, TNF-α and IL-6 levels, as well as the protein expressions of TGF-β, HIF-1α, α-SMA, cleaved-caspase-3, Jagged-1 and Notch-1 in renal tissue were significantly increased, while the serum IL-10 level, ATP content and activities of sodium-potassium ATPase and calcium-magnesium ATPase in mitochondria of renal tissue were significantly decreased (P<0.05). Compared with Model group, the renal tissue damage and fibrosis in the AST groups were reduced, the serum BUN and SCr levels, urine UP level, serum LDH, TNF-α and IL-6 levels, and the protein expressions of TGF-β, HIF-1α, α-SMA, cleaved-caspase-3, Jagged-1 and Notch-1 were significantly decreased, while the serum IL-10 level, ATP content and the activities of sodium-potassium ATPase and calcium-magnesium ATPase in mitochondria were significantly increased; the changes in the aforementioned indicators in AST- H group were more significant than those in the AST-L group(P<0.05). JFC could significantly reverse the improvement effect of high dose of AST on renal injury in CRF rats (P<0.05). CONCLUSIONS AST can reduce inflammation in CRF rats, alleviate renal tissue damage and fibrosis, and improve renal mitochondrial capacity metabolism, possibly by inhibiting the Jagged-1/Notch-1 signaling pathway.
3.Regulating ferroptosis of osteoblasts by traditional Chinese medicine in treatment of steroid-induced avascular necrosis of femoral head
Mianyu ZHANG ; Jie HAN ; Hao ZENG ; Xiangshan CHEN ; Zhengang GAO
Chinese Journal of Tissue Engineering Research 2025;29(1):185-192
BACKGROUND:Some studies have found that ferroptosis of osteoblasts can be an important mechanism to induce the occurrence and development of hormone-induced femoral head necrosis.With the development of Chinese medicine,some scholars have found that some Chinese medicine monomer,Chinese medicine compound and Chinese patent medicine can regulate the ferroptosis of osteoblasts through various pathway mechanisms,and finally play a role in the treatment of steroid-induced avascular necrosis of femoral head. OBJECTIVE:To investigate the relationship between ferroptosis and steroid-induced avascular necrosis of femoral head and the mechanism of Chinese medicine regulating ferroptosis of osteoblasts in the treatment of steroid-induced avascular necrosis of femoral head,so as to provide new ideas for the diagnosis and treatment of steroid-induced avascular necrosis of femoral head. METHODS:With"ferroptosis,steroid-induced avascular necrosis of femoral head,osteoblast,Chinese herbal medicine,glucocorticoid,iron metabolism,reactive oxygen species,glutathione peroxidase"as Chinese search terms,and"ferroptosis,hormonal necrosis of the femoral head,osteoblast,Chinese herbal medicine,glucocorticoid,iron metabolism,ROS,GPX4"as English search terms,the search was conducted on CNKI,PubMed,WanFang,VIP and other databases.The relevant articles on osteoblast ferroptosis and steroid-induced avascular necrosis of femoral head and the regulation of Chinese herbal medicine intervention from the establishment of each database to 2023 were screened.Finally,76 articles were systematically analyzed. RESULTS AND CONCLUSION:(1)Ferroptosis of osteoblasts plays an important role in the pathogenesis of steroid-induced avascular necrosis of femoral head.(2)The occurrence of ferroptosis in osteoblasts is regulated by a variety of mechanisms,such as intracellular iron overload causing ferroptosis.Lipid peroxidation damages cell membrane and causes ferroptosis.Cystine/glutamate reverse transporter induced ferroptosis by influencing glutathione level and glutathione peroxidase 4 activity.Fenton reaction in the cell produces a large number of reactive oxygen species and causes ferroptosis.(3)Chinese medicine monomer icariin,Chinese medicine compound Qinge pills and Chinese patent medicine Bushen Huoxue granules can regulate the occurrence of osteoblast ferroptosis,and help to prevent and treat steroid-induced avascular necrosis of femoral head.(4)The mechanism of ferroptosis in osteoblasts is still unclear.Further investigation on the mechanism of action of both is expected to provide a new choice for clinical treatment of steroid-induced avascular necrosis of femoral head.
4.Effect of electroacupuncture combined with low-frequency transcranial ultrasound stimulation on the electroencephalographic signals of rats with traumatic brain injury
Simiao GAO ; Xue HAN ; Xiaoguang WU ; Jinyu ZHENG ; Fangwen GAO ; Kuihua LI ; Yong PENG ; Lanxiang LIU
Chinese Journal of Tissue Engineering Research 2025;29(2):402-408
BACKGROUND:Traumatic brain injury is a condition in which the normal function of the brain is disrupted by a bump or impact to the head.It is necessary to find effective treatments and objective targets that can help doctors diagnose the injury status and restore the brain function of patients. OBJECTIVE:To explore the effect of electroacupuncture combined with low-frequency transcranial ultrasound stimulation on the electroencephalographic signals of rats with traumatic brain injury. METHODS:Forty 6-week-old SPF male Sprague-Dawley rats were randomly divided into five groups:sham group,model group,electroacupuncture group,low-frequency transcranial ultrasound stimulation group and combined group(electroacupuncture+low-frequency transcranial ultrasound stimulation),with eight rats in each group.Feeney weight-drop method was used to establish the animal model of traumatic brain injury.In the sham group,the bone window was only opened without impact.Interventions were started at 1 day after modeling.Electroacupuncture in the electroacupuncture group,low-frequency transcranial ultrasound stimulation in the low-frequency transcranial ultrasound stimulation group,and electroacupuncture+low-frequency transcranial ultrasound stimulation in the combined group were performed for days in total.The modified neurological severity scale score for assessing rats'neurological deficits was performed at 8 hours after modeling.The percentage of spontaneous alternation behavior in the Y-maze was measured at 7 days after modeling.Then,the electroencephalographic signals were collected and electroencephalographic data of α,β,θ,and δ waves were extracted by fast Fourier transform,and the value of oscillation amplitude and energy ratio were calculated in α,β,θ,and δ waves,as well as the Lempel-Ziv complexity and sample entropy. RESULTS AND CONCLUSION:Compared with the sham group,the modified neurological severity scale scores in the model group,electroacupuncture group,low-frequency transcranial ultrasound stimulation group and combined group were significantly increased at 8 hours after modeling(P<0.05).Compared with the sham group,the value of oscillation amplitude in δ wave and the value of δ energy ratio were significantly increased in the model group at 7 days after modeling,meanwhile the percentage of spontaneous alternation behavior in Y-maze,and the value of α/β energy ratio,Lempel-Ziv complexity,and sample entropy were significantly decreased(P<0.05).Compared with the model group,the value of oscillation amplitude in α and δ waves was significantly decreased in the combined group(P<0.05),while the value of α/β energy ratio was significantly increased(P<0.05)and the value of δ energy ratio was significantly decreased(P<0.05)in the electroacupuncture group,low-frequency transcranial ultrasound stimulation group and combined group.Compared with the electroacupuncture group and low-frequency transcranial ultrasound stimulation group,the value of δ energy ratio was significantly decreased in the combined group(P<0.05),while the percentage of spontaneous alternation behavior,the value of α/β energy ratio,the Lempel-Ziv complexity,and the sample entropy were significantly increased(P<0.05).To conclude,abnormal electroencephalographic signals can appear in rats with traumatic brain injury,while the electroacupuncture combined with low-frequency transcranial ultrasound stimulation can alleviate the abnormal electroencephalographic signals in rats,which suggests the electroencephalographic frequency domain value and nonlinear features can be used to assess the severity of traumatic brain injury.
5.Visual analysis of treatment of adolescent idiopathic scoliosis
Xiaodong ZHENG ; Shan GAO ; Wenjin HAN ; Lijun LIU ; Menglong JIA ; Longtan YU
Chinese Journal of Tissue Engineering Research 2025;29(3):645-653
BACKGROUND:At present,the incidence of scoliosis is increasing year by year,especially in adolescent idiopathic scoliosis.Therefore,it is more and more important to strengthen the research on the treatment of adolescent scoliosis. OBJECTIVE:To summarize the current status,hotspots,emerging trends,and frontiers of global research on the treatment of adolescent idiopathic scoliosis to provide reference and guidance for future related research. METHODS:The literature related to the treatment of adolescent idiopathic scoliosis was retrieved on the Web of Science Core Collection(WOSCC)database from 2013 to 2023.CiteSpace 6.2.R1 software was used for visual analysis of countries,institutions,authors,and keywords. RESULTS AND CONCLUSION:(1)A total of 561 English articles were included in this study.Among countries,institutions,and authors,the United States has contributed the most.Nanjing University and Qiu,Yong(Affiliated Drum Tower Hospital,Nanjing University School of Medicine)are the most published institution and author.The academic journal with the largest number of articles is the European Spine Journal.(2)In the analysis of cited literature,the top 10 most cited articles mainly describe the effects of surgical treatment and conservative treatment on improving adolescent idiopathic scoliosis,especially improving the curvature of patients.(3)Through the summary of highly cited articles and the keyword clustering,keyword prominence in-depth mining,the research hotspots are currently the relationship between Cobb angle and treatment choice,the therapeutic effect of exercise therapy and the therapeutic effect of posterior vertebral fusion.(4)The prognosis of patients with different curvatures has not been studied in depth,and the etiology of adolescent idiopathic scoliosis has not been clarified,so the relationship between curvature and prognosis and the etiology of adolescent idiopathic scoliosis may be a new research trend in the future.
6.Features of HBV RNA level in different stages of the natural history of chronic hepatitis B virus infection and its correlation with HBV DNA and HBsAg
Han GAO ; Juanli WU ; Yushuang ZHANG ; Yiheng ZHANG ; Lei WANG ; Tao LI ; Lixin ZHANG
Journal of Clinical Hepatology 2025;41(4):637-642
ObjectiveTo investigate the features of serum HBV RNA in different stages of the natural history of chronic hepatitis B virus (HBV) infection without antiviral treatment, as well as its correlation with serum HBV DNA and HBsAg. MethodsA total of 306 treatment-naïve patients with chronic HBV infection who attended Department of Infections Diseases and Hepatoloty, the Second Hospital of Shandong University from January 2023 to June 2024 were divided into six groups based on the different stages of natural history, i.e., HBeAg-positive chronic HBV infection group with 29 patients, HBeAg-positive chronic hepatitis B (CHB) group with 107 patients, HBeAg-negative chronic HBV infection group with 18 patients, HBeAg-negative CHB group with 60 patients, HBeAg-positive indeterminate-phase chronic HBV infection group with 7 patients, and HBeAg-negative indeterminate-phase chronic HBV infection group with 85 patients. Real-time isothermal RNA amplification was used to measure serum high-sensitivity HBV RNA. The Kruskal-Wallis H test was used for comparison between multiple groups of continuous data, while the Mann-Whitney U test was used for comparison between two groups. The Spearman method was used to investigate the correlation of HBV RNA with HBV DNA and HBsAg. ResultsThe HBeAg-positive chronic HBV infection group showed the highest level of serum HBV RNA [7.5 (7.4 — 7.9) log10 copies/mL], followed by the HBeAg-positive CHB group [7.4 (6.4 — 7.9) log10 copies/mL], the HBeAg-negative CHB group [4.5 (3.0 — 5.7) log10 copies/mL], and the HBeAg-negative chronic HBV infection group [1.0 (1.0 — 2.0) log10 copies/mL]; the HBeAg-positive indeterminate-phase chronic HBV infection group had a serum HBV RNA level of 3.9 (3.7 — 5.7) log10 copies/mL, and the HBeAg-negative indeterminate-phase chronic HBV infection group had a serum HBV RNA level of 2.0 (1.0 — 3.0) log10 copies/mL; there was a significant difference in serum HBV RNA level between the six groups (H=830.770, P<0.001). There was a significant difference in HBV RNA level between the HBeAg-positive chronic HBV infection group and all the other groups except the HBeAg-positive CHB group (all P<0.001). In the 306 patients with HBV infection, HBV RNA was strongly correlated with HBV DNA (r=0.92, P<0.001) and was moderately correlated with HBsAg (r=0.67, P<0.001). The correlation between serum HBV RNA and HBsAg in HBeAg-positive patients (r=0.61, P<0.001) was stronger than that in HBeAg-negative patients (r=0.31, P<0.001). For the patients with HBeAg-positive chronic HBV infection, the male patients with ALT>30 U/L and the female patients with ALT>19 U/L had a significantly lower serum HBV RNA level than the male patients with ALT≤30 U/L and the female patients with ALT≤19 U/L (P<0.001), and there was no significant difference in serum HBV RNA level between the latter group of patients and the HBeAg-positive CHB group (P>0.05). ConclusionIn patients with chronic HBV infection who do not receive antiviral therapy, there is a difference in serum HBV RNA level in different stages of natural history, and serum HBV RNA level has the strongest correlation with HBV DNA and a relatively weak correlation with HBsAg. In patients with HBeAg-positive chronic HBV infection, serum HBV RNA level in male patients with ALT>30 U/L and female patients with ALT>19 U/L are in the transition stage between HBeAg-positive chronic HBV infection and HBeAg-positive CHB.
7.History, Experience, Opportunities, and Challenges in Esophageal Cancer Prevention and Treatment in Linxian, Henan Province, A High Incidence Area for Esophageal Cancer
Lidong WANG ; Xiaoqian ZHANG ; Xin SONG ; Xueke ZHAO ; Duo YOU ; Lingling LEI ; Ruihua XU ; Jin HUANG ; Wenli HAN ; Ran WANG ; Qide BAO ; Aifang JI ; Lei MA ; Shegan GAO
Cancer Research on Prevention and Treatment 2025;52(4):251-255
Linxian County in Henan Province, Northern China is known as the region with the highest incidence and mortality rate of esophageal cancer worldwide. Since 1959, the Henan medical team has conducted field work on esophageal cancer prevention and treatment in Linxian. Through three generations of effort exerted by oncologists over 65 years of research on esophageal cancer prevention and treatment in Linxian, the incidence rate of esophageal squamous cell carcinoma in this area has dropped by nearly 50%, and the 5-year survival rate has increased to 40%, reaching the international leading
8.The mechanism of Laggerae Herba in improving chronic heart failure by inhibiting ferroptosis through the Nrf2/SLC7A11/GPX4 signaling pathway
Jinling XIAO ; Kai HUANG ; Xiaoqi WEI ; Xinyi FAN ; Wangjing CHAI ; Jing HAN ; Kuo GAO ; Xue YU ; Fanghe LI ; Shuzhen GUO
Journal of Beijing University of Traditional Chinese Medicine 2025;48(3):343-353
Objective:
To investigate the role and mechanism of the heat-clearing and detoxifying drug Laggerae Herba in regulating the nuclear factor-erythroid 2-related factor-2(Nrf2)/solute carrier family 7 member 11 (SLC7A11)/glutathione peroxidase 4 (GPX4) signaling pathway to inhibit ferroptosis and improve chronic heart failure induced by transverse aortic arch constriction in mice.
Methods:
Twenty-four male ICR mice were divided into the sham (n=6) and transverse aortic arch constriction groups (n=18) according to the random number table method. The transverse aortic arch constriction group underwent transverse aortic constriction surgery to establish models. After modeling, the transverse aortic arch constriction group was further divided into the model, captopril, and Laggerae Herba groups according to the random number table method, with six mice per group. The captopril (15 mg/kg) and Laggerae Herba groups (1.95 g/kg) received the corresponding drugs by gavage, whereas the sham operation and model groups were administered the same volume of ultrapure water by gavage once a day for four consecutive weeks. After treatment, the cardiac function indexes of mice in each group were detected using ultrasound. The heart mass and tibia length were measured to calculate the ratio of heart weight to tibia length. Hematoxylin and eosin staining were used to observe the pathological changes in myocardial tissue. Masson staining was used to observe the degree of myocardial fibrosis. Wheat germ agglutinin staining was used to observe the degree of myocardial cell hypertrophy. Prussian blue staining was used to observe the iron deposition in myocardial tissue. An enzyme-linked immunosorbent assay was used to detect the amino-terminal pro-brain natriuretic peptide (NT-proBNP) and glutathione (GSH) contents in mice serum. Colorimetry was used to detect the malondialdehyde (MDA) content in mice serum. Western blotting was used to detect the Nrf2, GPX4, SLC7A11, and ferritin heavy chain 1 (FTH1) protein expressions in mice cardiac tissue.
Results:
Compared with the sham group, in the model group, the ejection fraction (EF) and fractional shortening (FS) of mice decreased, the left ventricular end-systolic volume (LVESV) and left ventricular end-systolic diameter (LVESD) increased, the left ventricular anterior wall end-systolic thickness (LVAWs) and left ventricular posterior wall end-systolic thickness (LVPWs) decreased, the ratio of heart weight to tibia length increased, the myocardial tissue morphology changed, myocardial fibrosis increased, the cross-sectional area of myocardial cells increased, iron deposition appeared in myocardial tissue, the serum NT-proBNP and MDA levels increased, the GSH level decreased, and Nrf2, GPX4, SLC7A11, and FTH1 protein expressions in cardiac tissue decreased (P<0.05). Compared with the model group, in the captopril and Laggerae Herba groups, the EF, FS, and LVAWs increased, the LVESV and LVESD decreased, the ratio of heart weight to tibia length decreased, the myocardial cells were arranged neatly, the degree of myocardial fibrosis decreased, the cross-sectional area of myocardial cells decreased, the serum NT-proBNP level decreased, and the GSH level increased. Compared with the model group, the LVPWs increased, the iron deposition in myocardial tissue decreased, the serum MDA level decreased, and Nrf2, GPX4, SLC7A11, and FTH1 protein expressions in cardiac tissue increased (P<0.05) in the Laggerae Herba group.
Conclusion
Laggerae Herba improves the cardiac function of mice with chronic heart failure caused by transverse aortic arch constriction, reduces the pathological remodeling of the heart, and reduces fibrosis. Its mechanism may be related to Nrf2/SLC7A11/GPX4 pathway-mediated ferroptosis.
9.Effect of The Hydrophilic Amino Acids on Self-assembly Behavior of Short Bola-like Peptides
Xin-Xin GAO ; Yu HAN ; Yi-Lin ZHOU ; Xi-Ya CHEN ; Yu-Rong ZHAO
Progress in Biochemistry and Biophysics 2025;52(5):1290-1301
ObjectiveBola-like short peptides exhibit novel self-assembly properties due to the formation of peptide dimers via hydrogen bonding interactions between their C-terminals. In this configuration, hydrophilic amino acids are distributed at both terminals, making these peptides behave similarly to Bola peptides. The electrostatic repulsive interactions arising from the hydrophilic amino acids at each terminal can be neutralized, thereby greatly promoting the lateral association of β-sheets. Consequently, assemblies with significantly larger widths are typically the dominant nanostructures for Bola-like peptides. To investigate the effect of hydrophilic amino acids on the self-assembly behavior of Bola-like peptides, the peptides Ac-RI3-CONH2 and Ac-HI3-CONH2 were designed and synthesized using the Bola-like peptide Ac-KI3-CONH2 as a template. Their self-assembly behavior was systematically examined. MethodsAtomic force microscopy (AFM) and transmission electron microscopy (TEM) were employed to characterize the morphology and size of the assemblies. The secondary structures of the assemblies were analyzed using circular dichroism (CD) and Fourier transform infrared (FTIR) spectroscopy. Small-angle neutron scattering (SANS) was used to obtain detailed structural information at a short-length scale. Based on these experimental results, the effects of hydrophilic amino acids on the self-assembly behavior of Bola-like short peptides were systematically analyzed, and the underlying formation mechanism was explored. ResultsThe aggregation process primarily involved three steps. First, peptide dimers were formed through hydrogen bonding interactions between their C-terminals. Within these dimers, the hydrophilic amino acids K, R, and H were positioned at both terminals, enabling the peptides to self-assemble in a manner similar to Bola peptides. Next, β-sheets were formed via hydrogen bonding interactions along the peptide backbone. Finally, self-assemblies were generated through the lateral association of β-sheets. The results demonstrated that both Ac-KI3-CONH2 and Ac-RI3-CONH2 could self-assemble into double-layer nanotubes with diameters of approximately 200 nm. These nanotubes were formed by the edge fusion of helical ribbons, which initially emerged from twisted ribbons. Notably, the primary assemblies of these peptides exhibited opposite chirality: nanofibers formed by Ac-KI3-CONH2 displayed left-handed chirality, whereas those formed by Ac-RI3-CONH2 exhibited right-handed chirality. This reversal in torsional direction was primarily attributed to the different abilities of K and R to form hydrogen bonds with water. In contrast, Ac-HI3-CONH2 formed narrower twisted ribbons with a significantly reduced width of approximately 30 nm, which was attributed to the strong steric hindrance caused by the imidazole rings. The multilayer height of these ribbons was mainly due to the unique structure of the imidazole rings, which can function as both hydrogen bond donors and acceptors, thereby promoting aggregate growth in the vertical direction. ConclusionThe final morphology of the self-assemblies resulted from a delicate balance of various non-covalent interactions. By altering the types of hydrophilic amino acid residues in Bola-like short peptides, the relative strength of non-covalent interactions that drive assembly formation can be effectively regulated, allowing precise control over the morphology and chirality of the assemblies. This study provides a simple and effective approach for constructing diverse self-assemblies and lays a theoretical foundation for the development of functional biomaterials.
10.Re-examination of Atractylodis Rhizoma and Dosage of Whole Formula in Yuejiuwan
Yanping HAN ; Yiyi ZHANG ; Huimin GAO ; Raorao LI ; Li YAO ; Zhaoxiang SUN ; Zhuo MA ; Huamin ZHANG ; Wei ZHANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(13):223-233
Yuejuwan is a classic formula widely used by doctors to relieve liver and depression, with precise clinical efficacy in traditional Chinese medicine (TCM). The authors used bibliometric methods to collect and collate 495 ancient data related to Yuejuwan, and 105 valid data were screened out, involving 68 ancient Chinese medical books. After systematic verification of the origin of the formula of Yuejuwan, the main treatment symptoms, the principle of the formula, the composition of the drug, the dosage, the preparation method, the decoction method, and other information, the results showed that Yuejuwan originated from the Danxi Xinfa (《丹溪心法》) of the Yuan Dynasty by ZHU Zhenheng, and it is composed of five medicines, namely Atractylodis Rhizoma, Cyperi Rhizom, Chuanxiong Rhizoma, Massa Medicata Fermentata, and Gardeniae Fructus. In terms of drug base, Atractylodis Rhizoma, Cyperi Rhizom, Chuanxiong Rhizoma, and Gardeniae Fructus are in line with the records in the 2020 edition of Chinese Pharmacopoeia, and Massa Medicata Fermentata is used. The preparation method is as follows: Massa Medicata Fermentata and Gardeniae Fructus are fried, and Cyperi Rhizoma is roasted in vinegar. Chuanxiong Rhizoma is used in the raw form, and Atractylodis Rhizoma is prepared with rice swill. The formula can regulate Qi and relieve depression and broaden the middle and remove fullness. It is clinically used for the treatment of six types of depression syndromes, chest and diaphragm plumpness, abdominal distension and leg acid, acid swallowing and vomiting, eating and drinking disharmony, toothache, mouth and tongue sores, and other diseases. The most used dosage of the formula in the ancient records through the ages is converted into the modern dosage, namely 3.05 g Atractylodis Rhizoma, 3.05 g Cyperi Rhizoma, 3.05 g Chuanxiong Rhizoma, 3.05 g Massa Medicata Fermentata, and 3.05 g Gardeniae Fructus, and the daily dosage is 15.25 g. The converted dosage is similar to that recorded in the 2020 edition of the Chinese Pharmacopoeia. The formula is in pill form, and medicine should be taken with lukewarm boiled water after the meal. Through the excavation of the ancient literature related to Yuejuwan, the key information of the formula is identified, with a view to providing a more accurate reference for the clinical application of Yuejuwan and subsequent in-depth investigation.


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