1.The Effect of Zhiqiao Gancao Decoction (枳壳甘草汤) on Intervertebral Disc Nucleus Pulposus Cell Apoptosis and the Hippo-YAP/TAZ Signaling Pathway in Tail Intervertebral Disc Degeneration Model Rats
Zaishi ZHU ; Zeling HUANG ; Junming CHEN ; Bo XU ; Binjie LU ; Hua CHEN ; Xingxing DUAN ; Yuwei LI ; Xiaofeng SHEN
Journal of Traditional Chinese Medicine 2025;66(5):509-517
ObjectiveTo investigate the possible mechanism by which Zhiqiao Gancao Decoction (枳壳甘草汤, ZGD) delays intervertebral disc degeneration (IDD) based on the Hippo-yes-associated protein (YAP)/transcriptional co-activator with PDZ-binding motif (TAZ) signaling pathway. MethodsA total of 50 SD rats were randomly divided into sham surgery group, model group, low-dose ZGD group, high-dose ZGD group, and high-dose ZGD + inhibitor group, with 10 rats in each group. In the sham surgery group, the rats were pierced in the skin and muscle at the Co6/7/8 segments of the tail with a 21G needle (depth approximately 2 mm) without damaging the intervertebral disc. In the other groups, rats were injected with a 21G needle at the Co6/7/8 segments of the tail to establish an IDD model by piercing the tail intervertebral disc 5 mm. One week after modeling, rats in the low-dose and high-dose ZGD groups were given 6.24 and 12.24 g/(kg·d) of the decoction via gastric gavage, respectively. The high-dose ZGD + inhibitor group was given 12.24 g/(kg·d) of the decoction and an intraperitoneal injection of YAP/TAZ inhibitor Verteporfin 10 mg/kg. The sham surgery and model groups were given 5 ml/(kg·d) of normal saline via gavage. The gavage was given once a day, and the intraperitoneal injection was given every other day. After 4 weeks of continuous intervention, the pathological changes of the tail intervertebral discs were observed using HE staining, Oil Red O-Green staining, and Toluidine Blue staining. Immunohistochemistry was used to detect the expression of aggrecan and MMP3 in the nucleus pulposus. TUNEL fluorescence staining was performed to detect apoptosis in the nucleus pulposus, and the apoptosis rate was calculated. Western blot was used to detect the Hippo-YAP/TAZ signaling pathway, including YAP, phosphorylated YAP (p-YAP), phosphorylated MST1/2 (p-MST1/2), phosphorylated TAZ (p-TAZ) and apoptosis-related proteins, such as Cleaved Caspase 3, P53, Bcl-2 and Bax. ResultsCompared with sham surgery group, the rats in the model group showed significant degenerative changes in the intervertebral disc. The levels of aggrecan, Bcl-2, and YAP proteins in the nucleus pulposus decreased, while the levels of p-MST1/2, p-YAP, p-TAZ, P53, Bax, Cleaved Caspase 3, MMP3 proteins, and the apoptosis rate increased (P < 0.01). Compared with the model group, the drug intervention groups showed partial recovery in intervertebral disc degeneration. The levels of aggrecan, Bcl-2, and YAP proteins increased, while the levels of p-MST1/2, p-YAP, p-TAZ, P53, Bax, Cleaved Caspase 3, MMP3 proteins, and the apoptosis rate decreased (P<0.05 or P<0.01). The high-dose ZGD group showed more significant recovery in intervertebral disc degeneration compared to the low-dose ZGD group, with a decrease in the levels of p-MST1/2, p-YAP, p-TAZ, P53, Bax, Cleaved Caspase 3, MMP3 proteins, and apoptosis rate, and an increase in the levels of aggrecan, Bcl-2, and YAP proteins (P<0.05 or P<0.01). Compared with the high-dose ZGD group, the high-dose ZGD + inhibitor group showed a reduced recovery in intervertebral disc degeneration, with an increase in the levels of p-MST1/2, p-YAP, p-TAZ, P53, Bax, Cleaved Caspase 3, MMP3 proteins, and apoptosis rate, and a decrease in the levels of aggrecan, Bcl-2, and YAP proteins (P<0.05 or P<0.01). ConclusionZGD may delay intervertebral disc degeneration by inhibiting the phosphorylation of YAP in the nucleus pulposus, maintaining the function of the Hippo-YAP/TAZ signaling pathway, and reducing apoptosis of nucleus pulposus cells.
2.Timosaponin BⅡ Combined with Icariin Maintains Osteoclast-osteoblast Coupling by Restoring Yin-Yang Balance
Zaishi ZHU ; Zeling HUANG ; Weiye CAI ; Hua CHEN ; Boen SONG ; Yue LU ; Qing LU ; Xiaofeng SHEN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(18):48-57
ObjectiveTo explore the effect of timosaponin BⅡ (TBⅡ) combined with icariin (ICA) on osteoclast (OC)-osteoblast (OB) coupling and decipher the mechanism from the cellular level. MethodsThe cell counting kit-8 (CCK-8) was used to assess the effects of different concentrations of TBⅡ and different concentrations of TBⅡ+ICA on the growth of RAW264.7 cells. Soluble receptor activator of nuclear factor-κB ligand (sRANKL) was used to induce the differentiation of RAW264.7 pre-osteoclasts into osteoclasts. The cells were allocated into sRANKL, TBⅡ (1, 5, 10 μmol·L-1), and TBⅡ+ICA groups. Tartrate-resistant acid phosphatase staining was performed to assess the effects of TBⅡ and TBⅡ+ICA on osteoclast differentiation. Real-time quantitative polymerase chain reaction (Real-time PCR) was conducted to examine the effects of TBⅡ+ICA on the expression of key genes involved in osteoclast differentiation and osteoclast-derived coupling factors. The osteogenic differentiation conditioned medium mixed with osteoclast supernatant was used to induce osteogenic differentiation of MC3T3-E1 cells. Alkaline phosphatase staining and alizarin red S staining were employed to determine the effect of TBⅡ+ICA on osteogenic differentiation. Real-time PCR was employed to evaluate the effects of conditioned medium on key genes involved in osteogenic differentiation. ResultsTBⅡ at 1, 5, 10 μmol·L-1 had no significant effect on the cell survival rate. Compared with the sRANKL group, TBⅡ inhibited osteoclast differentiation in a dose-dependent manner and achieved the best effect at 10 μmol·L-1 (P<0.01). Compared with the sRANKL group, different concentrations of TBⅡ down-regulated the mRNA levels of osteoclast differentiation-related genes c-Fos, RANK, and RANKL (P<0.05). None of 10 μmol·L-1 TBⅡ, 10 μmol·L-1 TBⅡ+10-4 μmol·L-1 ICA, or 10 μmol·L-1 TBⅡ+10-3 μmol·L-1 ICA affected the viability of RAW264.7 cells. TBⅡ and/or ICA inhibited osteoclast differentiation (P<0.01), and TBⅡ + ICA had the best effect (P<0.01). Compared with the sRANKL group, TBⅡ and/or ICA down-regulated the mRNA levels of c-Fos, RANK, and RANKL (P<0.05). The single application of TBⅡ and ICA had no significant effect on the mRNA levels of Wnt10b, Cthrc1, and C3a, while TBⅡ+ICA exerted up-regulating effects (P<0.05). Compared with those in the blank group, the bone differentiation and mineralization abilities of the normal osteogenic induction group and each osteogenic induction + osteoclast supernatant group were improved (P<0.01). Compared with the blank group, the normal osteogenic induction group and the osteogenic induction + osteoclast supernatant group showed up-regulated mRNA levels of Runx2 and OCN (P<0.01). ConclusionTBⅡ+ICA can inhibit osteoclast differentiation, maintain the normal osteoclast-osteoblast coupling, and promote osteogenic differentiation.
3.Ultra-long-distance single-port robotic-assisted laparoscopic renal cyst decortication using a domestic robotic system:a case report
Xiaofeng WU ; Liru CHEN ; Yu FENG ; Wei ZHANG ; Linhui WANG ; Xiaoying LU
Academic Journal of Naval Medical University 2025;46(11):1510-1513
Objective To summarize the operation experience of 1 case of ultra-long-distance single-port robotic-assisted laparoscopic renal cyst decortication using a domestic robotic system,and provide references for the development of long-distance telesurgery.Methods The procedure was performed using a domestically produced single-port robotic system.The distance between the main control end and the patient end was 2 400 km.The operation was conducted remotely via a 5G network and a 100 Mbps dedicated line provided by China Telecom.The surgeon controlled the robotic arm remotely.Results The surgery was completed smoothly within a duration of 32 min.The real-time latency was(90±20)ms.Blood loss during the operation was 10 mL,and no drainage tube was placed.The patient recovered smoothly and returned to the ward without any complications.Conclusion It is safe and feasible for the domestically produced single-port robotic system to implement ultra-long-distance telesurgery.This approach facilitates the decentralization of advanced medical resources and helps mitigate disparities in healthcare access.
4.Preparation of doxorubicin-loaded polyphyllin H liposomes and synergistic anti-tumor activity against non-small cell lung cancer in vitro
Yining LIU ; Dawei ZHOU ; Shouchang GAI ; Lu SUI ; Xue SUN ; Zhenhua TONG ; Yuhang WANG ; Jing ZHAO ; Xiaofeng YUAN ; Yong XIANG
Journal of Army Medical University 2025;47(17):2134-2144,封3
Objective To prepare glucose transporter 1(Glut1)-targeted doxorubicin(DOX)-loaded liposomes(doxorubicin/polyphyllin H-liposomes,DOX/ppH-LPs)using polyphyllin H(ppH)instead of cholesterol as the liposomal membrane material,and to investigate their in vitro synergistic anti-tumor activity against non-small cell lung cancer(NSCLC).Methods DOX/ppH-LPs were prepared using thin-film hydration,and the formulation was optimized by single-factor investigation.The optimized DOX/ppH-LPs were characterized for morphology,particle size,polydispersity index(PDI),and zeta potential with transmission electron microscopy(TEM)and dynamic light scattering(DLS).Drug loading DL%was determined by high-performance liquid chromatography(HPLC).The storage stability was evaluated by observing in PBS at 4℃for 7 d,and the serum stability was observed in DMEM containing 10%fetal bovine serum(FBS)at 37℃for 48 h.In vitro drug release was studied in PBS at pH 7.4 and pH 5.0 values,respectively.Human NSCLC A549 cells were subjected as the model,MTT assay was performed to detect the proliferation inhibition by DOX/ppH-LPs at different concentrations(0.5,5.0,15.0 μg/mL)and the control group(ppH+DOX/LPs,a physical mixture of free ppH and DOX-loaded liposomes).Fluorescence microscopy was used to observe cellular uptake of DOX/ppH-LPs and DOX/LPs(containing 5 μg/mL DOX)at 15 min and 2 h.Live/dead cell staining was applied to assess apoptosis/necrosis induced by formulations(15 μg/mL DOX)after 48 h incubation.Transwell assay was conducted to evaluate inhibitory effect on cell migration and invasion,and the targeting property and in vitro synergistic anti-NSCLC activity of DOX/ppH-LPs were then comprehensively evaluated.Results The optimal formulation of DOX/ppH-LPs was determined as hydration temperature at 50℃,6 mg DOX,2 mg ppH,and 24 mg lecithin.The prepared DOX/ppH-LPs were in spherical shape,uniform distribution,and at an average particle size of 145.13±22.14 nm,a PDI of 0.15±0.05,a zeta potential of-23.92±1.73 mV,and a DL of 10.13±0.71%for DOX and(1.22±0.21)%for ppH.DOX/ppH-LPs maintained stable particle size,PDI,and exhibited significantly unchanged zeta potential after storage in PBS at 4℃for 7 d or incubation in DMEM containing 10%FBS at 37℃for 48 h,demonstrating excellent physical and serum stability.Both liposomes showed slow release at pH 7.4 value,while drug release was significantly accelerated at pH 5.0 value(P<0.05),indicating pH-sensitive release characteristics.MTT assay revealed that DOX/ppH-LPs exerted significantly stronger cytotoxicity against A549 cells than the ppH+DOX/LPs control group(P<0.05).Compared with ppH+DOX/LPs,DOX/ppH-LPs showed remarkably enhanced cellular uptake in A549 cells(P<0.05),with more DOX localized in the nucleus.Live/dead cell staining showed that at the same DOX concentration(15 μg/mL),the proportion of apoptotic/necrotic cells induced by DOX/ppH-LPs was significantly higher than that of the DOX/LPs control group.Transwell assay demonstrated that there were significantly less cells migrating and invading through the membrane in the DOX/ppH-LPs group than the ppH+DOX/LPs group.Conclusion Glut1-targeted doxorubicin-loaded liposomes(DOX/ppH-LPs)constructed by substituting cholesterol with ppH can target NSCLC cells,significantly enhance the in vitro synergistic anti-NSCLC activity of DOX and ppH.
5.Diagnostic value of preoperative diffusion weighted imaging histogram parameters in the depth of invasion of early rectal cancer
Shengchao JI ; Xiaofeng JIN ; Daixi YE ; Zehua LU ; Lulu XUAN ; Chengjun GENG
Journal of International Oncology 2025;52(10):621-627
Objective:To explore the diagnostic value of preoperative diffusion weighted imaging (DWI) histogram parameters in the depth of invasion of early rectal cancer.Methods:A total of 180 patients with early rectal cancer admitted to 904th Hospital of Joint Logistics Support Force of Chinese People's Liberation Army from August 2020 to August 2024 were selected as the study objects. Patients were divided into intramucosal cancer group ( n=102) and submucosal cancer group ( n=78) according to the depth of tumor invasion. The general data of the two groups were compared. The intraclass correlation coefficient (ICC) was used to analyze the consistency of DWI histogram parameters extracted by the two radiologists, and the differences between the two groups were compared. Receiver operator characteristic (ROC) curve was used to analyze the predictive value of each parameter to the depth of tumor invasion. Multivariate logistic regression was used to analyze the independent influencing factors of invasion depth, and a predictive model was constructed. The ROC curve was drawn to analyze the predictive value of the model for tumor invasion depth, and the Hosmer-Lemeshow test was used to analyze the goodness of fit of the model. Results:There were statistically significant differences in age ( t=8.15, P<0.001), maximum tumor diameter ( χ2=29.29, P<0.001), endoscopic type ( χ2=20.96, P<0.001), histological type ( χ2=24.93, P<0.001) and differentiation degree ( χ2=73.35, P<0.001) between intramucosal cancer group and submucosal cancer group. The mean, variance, skewness, kurtosis, the 1 st, 10 th, 50 th, 90 th, and 99 th percentiles of the histogram parameters of DWI had good consistency (all ICC>0.75). There were statistically significant differences in the mean ( t=5.69, P<0.001), variance ( t=9.75, P<0.001), skewness ( t=10.88, P<0.001), kurtosis ( t=10.06, P<0.001), the 1 st percentile ( t=3.43, P<0.001), 10 th percentile ( t=3.59, P<0.001), 50 th percentile ( t=9.97, P<0.001), 90 th percentile ( t=4.63, P<0.001), and 99 th percentile ( t=2.44, P=0.016) of the DWI histogram parameters between the intramucosal cancer group and the submucosal cancer group. ROC curve analysis results showed that mean [area under the curve (AUC) =0.77], variance (AUC=0.88), skewness (AUC=0.88), kurtosis (AUC=0.78), 50 th percentile (AUC=0.86) and 90 th percentile (AUC=0.82) had certain diagnostic value for submucous cancer. Multivariate analysis showed that age ( OR=9.98, 95% CI: 1.10-90.70, P=0.041), maximum tumor diameter ( OR=7.36, 95% CI: 1.08-50.23, P=0.042), and differentiation degree ( OR=19.88, 95% CI: 1.21-327.92, P=0.037), variance ( OR=16.24, 95% CI: 2.26-116.68, P=0.006), skewness ( OR=21.13, 95% CI: 2.80-59.61, P=0.003), 1 st percentile ( OR=9.78, 95% CI: 1.17-81.76, P=0.035) were independent factors in predicting tumor invasion depth in patients with early rectal cancer. The predictive model based on the above indicators was logit ( P) =1.51+2.30×age+2.00×maximum tumor diameter+2.99×differentiation degree+2.79×variance+3.05×skewness+ 2.28×the 1 st percentile. ROC curve analysis showed that the predictive model had an AUC of 0.97 (95% CI: 0.95-0.99) for judging the occurrence of submucosal cancer in patients with early rectal cancer, the sensitivity was 0.95, and the specificity was 0.88. The Hosmer-Lemeshow test results showed that the goodness of fit of the model was ideal ( P=0.823) . Conclusions:Age, maximum tumor diameter, differentiation degree, variance, skewness, and the 1 st percentile are independent factors in predicting tumor invasion depth in patients with early rectal cancer. The predictive model constructed based on these factors can effectively predict the risk of submucosal cancer in patients with early rectal cancer.
6.High expression of CPNE3 correlates with poor long-term prognosis of gastric cancer by inhibiting cell apoptosis via activating PI3K/AKT signaling
Ting DUAN ; Zhen ZHANG ; Jinran SHI ; Linyu XIAO ; Jingjing YANG ; Lixia YIN ; Xiaofeng ZHANG ; Zhijun GENG ; Guoyu LU
Journal of Southern Medical University 2024;44(1):129-137
Objective To explore the correlation of CPNE3 expression with long-term prognosis of patients with gastric cancer(GC)and the possible mechanism.Methods We retrospectively collected the data of 104 GC patients undergoing radical surgery in our hospital from February,2013 to October,2017.TCGA database and immunohistochemistry were used to analyze CPNE3 expression level in GC tissues and its effects on tumor progression and long-term prognosis of the patients.GO analysis was performed to predict the biological role of CPNE3 in GC.We also conducted cell experiments with MGC803 cells and observed the effects of CPNE3 knockdown,CPNE3 overexpression and LY294002(a PI3K/AKT inhibitor)treatment on cell apoptosis and cellular expressions of apoptotic proteins using flow cytometry and Western blotting.Results TCGA analysis and immunohistochemistry both showed high expressions of CPNE3 in GC(P<0.05).The patients with high CPNE3 expressions had a reduced 5-year survival(P<0.01),and a high CPNE3 expression,CEA level≥5 μg/L,CA19-9 level≥37 kU/L,T3-T4 stage,and N2-N3 stage were all independent risk factors for a lowered 5-year survival rate after surgery.The sensitivity and specificity of CPNE3 for predicting 5-year mortality was 79.59%and 74.55%,respectively(P<0.05).GO analysis predicted that CPNE3 negatively regulated GC cell apoptosis.In MGC803 cells,CPNE3 knockdown significantly increased cell apoptosis,enhanced Bax and Cleaved Caspase-3 expressions and decreased Bcl-2 expression,while CPNE3 overexpression produced the opposite results(P<0.05).The cellular expressions of p-PI3K and p-AKT were significantly decreased following CPNE3 knockdown and increased following CPNE3 overexpression(P<0.05).Treatment with LY294002 obviously attenuated the inhibitory effect of CPNE3 overexpression on apoptosis of MGC803 cells(P<0.05).Conclusion CPNE3 is highly expressed in GC tissues and affects the long-term prognosis of the patients possibly by inhibiting GC cell apoptosis through activation of PI3K/AKT signaling.
7.Role of Abelson interactor 2 in progression and prognosis of gastric cancer and its regulatory mechanisms
Xiaohua CHEN ; Hui LU ; Ziliang WANG ; Lian WANG ; Yongsheng XIA ; Zhijun GENG ; Xiaofeng ZHANG ; Xue SONG ; Yueyue WANG ; Jing LI ; Jianguo HU ; Lugen ZUO
Journal of Southern Medical University 2024;44(9):1653-1661
Objective To explore the regulatory role of Abelson interactor 2(ABI2)in progression and prognosis of gastric cancer.Methods TIMER2.0,GEPIA,Kaplan-Meier Plotter and DAVID databases were used to analyze ABI2 expression in pan-cancer and its association with the prognosis of gastric cancer.Gastric cancer and adjacent tissues from 120 patients undergoing radical gastrectomy in our hospital between January,2016 and October,2018 were examined for ABI2 expression and its correlation with disease progression and prognosis.MGC-803 cell models of ABI2 knockdown and overexpression were established for observing the changes in cell proliferation,migration,and invasion,and the impact of ABI2 expression modulation on xenograft growth was evaluated in nude mice.Results Database analysis and examination of the clinical samples showed that ABI2 was highly expressed in gastric cancer tissues.Survival analysis suggested that gastric cancer patients with a high expression of ABI2 had a reduced postoperative 5-year survival rate(P<0.0001),and further Cox univariate and multivariate survival analyses indicated that a high ABI2 expression was an independent risk factor affecting the patients survival outcomes(P=0.022,HR=1.887,95%CI:1.096-3.249).Enrichment analysis suggested the involvement of ABI2 in Wnt signaling.In MGC-803 cells,ABI2 overexpression promoted cell proliferation and xenograft growth in nude mice,increased the expressions of vimentin and N-cadherin,and lowered E-cadherin expression,while ABI2 knockdown produced the opposite effects.Mechanistic analysis revealed that ABI2 overexpression promoted the expressions of Wnt2 and β-catenin in both MGC-803 cells and the xenografts,and their expressions were significantly lowered by ABI2 knockdown.Conclusion ABI2 is highly expressed in gastric cancer,which affects long-term prognosis of the patients,possible due to its regulatory effect on Wnt signaling to promote proliferation,migration and invasion of gastric cancer cells.
8.Comparison of clinical features and outcomes of proliferative, fibrotic, and mixed subtypes of IgG4-related disease: A retrospective cohort study
Linyi PENG ; Xinlu ZHANG ; Jiaxin ZHOU ; Jieqiong LI ; Zheng LIU ; Hui LU ; Yu PENG ; Yunyun FEI ; Yan ZHAO ; Xiaofeng ZENG ; Wen ZHANG
Chinese Medical Journal 2024;137(3):303-311
Background::Immunoglobulin G4-related disease (IgG4-RD) is a recently recognized immune-mediated disorder that can affect almost any organ in the human body. IgG4-RD can be categorized into proliferative and fibrotic subtypes based on patients’ clinicopathological characteristics. This study aimed to compare the clinical manifestations, laboratory findings, and treatment outcomes of IgG4-RD among different subtypes.Methods::We prospectively enrolled 622 patients with newly diagnosed IgG4-RD at Peking Union Medical College Hospital from March 2011 to August 2021. The patients were divided into three groups according to their clinicopathological characteristics: proliferative, fibrotic, and mixed subtypes. We compared demographic features, clinical manifestations, organ involvement, laboratory tests, and treatment agents across three subtypes. We then assessed the differences in treatment outcomes among 448 patients receiving glucocorticoids alone or in combination with immunosuppressants. Moreover, risk factors of relapse were revealed by applying the univariate and multivariate Cox regression analysis.Results::We classified the 622 patients into three groups consisting of 470 proliferative patients, 55 fibrotic patients, and 97 mixed patients, respectively. We found that gender distribution, age, disease duration, and frequency of allergy history were significantly different among subgroups. In terms of organ involvement, submandibular and lacrimal glands were frequently involved in the proliferative subtype, while retroperitoneum was the most commonly involved site in both fibrotic subtype and mixed subtype. The comparison of laboratory tests revealed that eosinophils ( P = 0.010), total IgE ( P = 0.006), high-sensitivity C-reactive protein ( P <0.001), erythrocyte sedimentation rate ( P <0.001), complement C4 ( P <0.001), IgG ( P = 0.001), IgG1 (P <0.001), IgG4 (P <0.001), and IgA ( P <0.001), at baseline were significantly different among three subtypes. Compared with proliferative and mixed subtypes, the fibrotic subtype showed the lowest rate of relapse (log-rank P = 0.014). Conclusions::Our study revealed the differences in demographic characteristics, clinical manifestations, organ involvement, laboratory tests, treatment agents, and outcomes across proliferative, fibrotic, and mixed subtypes in the retrospective cohort study. Given significant differences in relapse-free survival among the three subtypes, treatment regimens, and follow-up frequency should be considered separately according to different subtypes.Trial Registration::ClinicalTrials. gov, NCT01670695.
9.Effective Components and Antiarrhythmic Mechanisms of Wenxin Granules Based on CMC/UPLC-Q-TOF/MS
Lu YU ; Shule QIAN ; Haizhen GUO ; Yuke ZHAO ; Xiaofeng LI ; Wuxun DU
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(19):124-132
ObjectiveTo employ the effective components and antiarrhythmic mechanism of Wenxin Granules (WXKL) by cell membrane chromatography (CMC) and ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS), combined with network pharmacology. MethodIn this study, the CMC/UPLC-Q-TOF/MS technique was employed to identify the components in WXKL that could specifically bind to myocardial cell membranes. By utilizing databases such as SwissTarget Prediction and GeneCards, the targets of WXKL's effective components and arrhythmia-related targets were mined. Cytoscape software was used to construct a "component-target-disease" network. Gene ontology(GO) function and Kyoto encyclopedia of genes and genomes(KEGG) pathway enrichment analyses were carried out, and molecular docking of key components and targets was performed. Finally, further verification was conducted through in vivo experiment of rats. ResultA total of 39 effective components were identified in WXKL. These included 13 components derived from Panax notoginseng, 15 components from Codonopsis pilosula, seven components from Glycyrrhizae Radix et Rhizoma, one component from Succinum, one component from Polygonatum odoratum, one component shared by both P. odoratum and C. pilosula, and one component shared by both Panax notoginseng and C. pilosula. Network pharmacology predicted that WXKL had 16 core antiarrhythmic targets and 79 related pathways, mainly involving adrenergic signaling in cardiomyocytes, cyclic guanosine monophosphate (cGMP)/protein kinase G (PKG), calcium signal, cyclic adenosine monophosphate (cAMP), interleukin (IL)-17, mitogen-activated protein kinase (MAPK), and tumor necrosis factor (TNF) signaling pathways. The results of in vivo experiment of rats showed that WXKL significantly improved the expression of β1-adrenergic receptor (β1-AR), cAMP, TNF-α, and calcium voltage-gated channel subunit alpha 1C (CACNA1C). ConclusionWXKL can exert its antiarrhythmic effects through multiple components, multiple targets, and multiple pathways. This study provides a scientific basis for explaining the potential pharmacodynamic substance foundation and mechanism of action of traditional Chinese medicine in treating arrhythmia.
10.Effects of marathon exercise on knee cartilage volume and T2 relaxation time
Lingbin XU ; Feng FU ; Xiaofeng YANG ; Qiqian SANG ; Yafei XU ; Mingjie WU ; Lu XUE
Chinese Journal of Orthopaedics 2024;44(5):294-301
Objective:To investigate the effects of marathon exercise on knee cartilage volume and T2 relaxation time (T2 value) based on MRI.Methods:From December 2018 to December 2021, 25 healthy volunteers without long-distance running habits and 32 non-professional marathon runners with long-term long-distance running were recruited to undergo knee MRI 3D water-selective excitation (three dimensional water-selective excitation, 3D-WATS) and T2 mapping imaging were performed, and the cartilage volumes in 5 knee areas and T2 values in 42 subareas were extracted for analysis. To compare the cartilage volume and its ratio to body surface area of knee joint of healthy volunteers and non-professional marathon runners, the T2 value of cartilage in each subregion, and the correlation between marathon exercise intensity and the volume and T2 value of cartilage in different regions.Results:Compared with healthy volunteers, there was no significant difference in cartilage volume or the ratio of body surface area to body volume of non-professional marathon runners ( P>0.05). There were significant differences between healthy volunteers and non-professional marathon runners in cartilage T2 values of the median layer of medial condyle of femur (47.61±5.65 ms and 44.29±6.10 ms) and the deep layer of medial condyle of femur (36.82±9.05 ms and 31.67±7.59 ms), deep precondylar area of medial femur (38.37±4.68 ms and 34.09±4.19 ms), shallow area of medial condylar area of femur (52.17±11.11 ms and 45.51±7.76 ms), middle layer of medial condylar area of femur (49.09±5.08 ms and 45.63±5.04 ms), medial layer of anterior condylar region of lateral femur (45.69±4.68 ms and 42.57±5.77 ms), superficial layer of posterior condylar region of lateral femur (55.42±18.41 ms and 47.99±8.39 ms), deep layer of anterior tibial medial plateau (33.40±7.76 ms and 29.03±5.69 ms), deep layer of posterior tibial medial plateau (31.28±5.02 ms and 27.92±5.99 ms), deep layer of patellofemoral surface (35.65±6.99 ms and 32.30±5.28 ms), respectively ( P<0.05). In non-professional marathon runners, the medial tibial plateau cartilage volume was negatively correlated with step frequency ( r=-0.371, P=0.035), the lateral femoral condylar cartilage volume was negatively correlated with step frequency ( r=-0.365, P=0.043), and the lateral tibial plateau cartilage volume was negatively correlated with step frequency ( r=-0.550, P=0.001). The T2 value of the medial layer cartilage in the anterior tibial medial plateau region was negatively correlated with body weight ( r=-0.277, P=0.039) and body mass index ( r=-0.290, P=0.030). The T2 value of the superficial layer of patellofemoral surface was negatively correlated with the amount of running in 3 months ( r=-0.457, P=0.010). The superficial T2 value in the posterior lateral plateau of the tibia was negatively correlated with stride length ( r=-0.437, P=0.014), and the medial layer cartilage T2 value in the anterior condylar area of the lateral femur was negatively correlated with stride frequency ( r=-0.380, P=0.035). Conclusion:Marathon exercise had little effect on the knee cartilage volume, but had a certain effect on the cartilage T2 value, resulting in changes in cartilage structure. The higher the step frequency, the smaller the cartilage volume. The greater the body weight or body mass index, the greater the amount of running in 3 months, and the greater the stride length, the lower the cartilage T2 value.

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