OBJECTIVES: To evaluate the protective effect of Bufei Yishen Formula (BYF) against cigarette smoke extract (CSE)-induced injuries in human bronchial epithelial BEAS-2B cells and explore the underlying mechanism. METHODS: BEAS-2B cells exposed to CSE were treated with normal rat serum, BYF-medicated rat serum at low or high doses, pyrrolidine dithiocarbamate (PDTC, a NF-κB inhibitor), PDTC combined with high-dose BYF-medicated serum, or S-carbomethyloysteine (S-CMC, as the positive control). CCK-8 assay was used to determine the optimal concentration and treatment time of CSE, BYF-medicated serum and S-CMC. The treated cells were examined for inflammatory factor levels in the supernatant and cellular expressions of MUC5AC and MUC5B using ELISA, cell ultrastructural changes with transmission electron microscopy, and cell apoptosis rate using flow cytometry. The expression levels of TLR4/NF‑κB pathway-associated mRNAs and proteins were determined by qRT-PCR and Western blotting. RESULTS: CSE exposure significantly increased secretions of IL-1β, IL-6 and TNF-α, mRNA and protein expressions of MUC5AC and MUC5B, and early and total apoptosis rates in BEAS-2B cells, where the presence of apoptotic bodies was detected. CSE also significantly enhanced the mRNA and protein expressions of TLR4, I-κB, and NF-κB and reduced mRNA and protein expressions of AQP5. Treatments of the CSE-exposed cells with BYF-medicated serum, PDTC and S-CMC all significantly lowered inflammatory factor levels, MUC5AC and MUC5B expressions, and early and total cell apoptosis rates, and partly reversed the changes in cellular ultrastructure and mRNA and protein expressions of the TLR4/NF-κB pathway, and the effects were the most conspicuous following the combined treatment with high-dose BYF-medicated serum and PDTC. CONCLUSIONS BYF can inhibit cell apoptosis, inflammation and mucus hypersecretion in CSE-induced BEAS-2B cells by inhibiting the TLR4/NF-κB signaling pathway.
Humans ; Epithelial Cells/cytology* ; Drugs, Chinese Herbal/pharmacology* ; NF-kappa B/metabolism* ; Bronchi/cytology* ; Smoke/adverse effects* ; Apoptosis/drug effects* ; Mucin 5AC/metabolism* ; Cell Line ; Toll-Like Receptor 4/metabolism* ; Mucin-5B/metabolism* ; Signal Transduction/drug effects* ; Nicotiana ; Rats ; Thiocarbamates/pharmacology* ; Animals

AIM:This study aimed to explore the mechanism by which Bufei-Yishen formula(BYF)mitigates mitochondrial damage in rats with chronic obstructive pulmonary disease(COPD)by regulating the AMPK/PGC-1α signal-ing pathway.METHODS:Forty rats were randomly divided into four groups,each containing ten rats each:control group,COPD group,BYF group,and N-acetylcysteine(NAC)group.The COPD model was established through chronic cigarette smoke exposure combined with periodic bacterial inoculations over an eight-week induction phase.During the subsequent eight-week treatment period(i.e.,weeks 9～16),rats in the control and COPD groups received an isovolumet-ric saline solution via oral gavage,at a standardized daily dose of 2 mL per animal.Moreover,rats in the BYF and NAC groups were given Bufei Yishen formula(11.61 g·kg-1·d-1)or N-acetylcysteine(54 mg·kg-1·d-1)by gavage,once per day.At week 16,samples were collected and the general condition of the rats was observed.Body weight was recorded weekly.We also obtained data characterizing rat lung function,lung pathology,ATP content,and mitochondrial ultra-structure,as well as the levels of interleukin-6(IL-6),tumor necrosis factor-alpha(TNF-α),interleukin-1β(IL-1β),se-rum transforming growth factor-beta 1(TGF-β1)and the enzymatic activities of mitochondrial electron transport chain complexes I(NADH dehydrogenase)and III(cytochrome c reductase).Finally,we quantified the mRNA and protein lev-els of AMPK and PGC-1α in lung tissue.RESULTS:Compared to the control group,the COPD group exhibited yellow-ish hair color,reduced gloss,slower weight gain,and a disordered respiratory rhythm.We also observed significant de-creases(P<0.01)in pulmonary function tidal volume(TV),minute ventilation(MV),peak expiratory flow(PEF),expi-ratory flow at 50%of tidal volume(EF50),forced vital capacity(FVC),forced expiratory volume in 0.1 s(FEV0.1),and FEV0.1/FVC.Histopathological analysis showed alveolar cavity enlargement,bullous changes in lung morphology,smooth muscle hypertrophy in the tracheal wall,ciliary destroyed,mucosal shrinking and thickening,and a large number of in-flammatory cells gathered around the tube.Moreover,the mean linear intercept(MLI)and bronchial wall thickness(BWt)had both significantly increased(P<0.01).Electron microscopic analysis of the lungs revealed a reduction in the number of mitochondria in alveolar epithelial cells,a swollen and deformed lung morphology overall.We observed that the mitochondrial cristae were broken,dissolved or vacuolated,accompanied by a significant reduction in the number of lamel-lar bodies and lung volume,along with a disordered internal lipid layer structure.Furthermore,some lung samples were vacuolated or had content leakage.Further quantitative analyses showed statistically significant increases(P<0.01)in the levels of serum pro-inflammatory mediators,including IL-6,TNF-α,IL-1β,and TGF-β1.At the same time we observed substantial reductions in the enzymatic activities of mitochondrial electron transport chain complexes I and III(P<0.01).Moreover,we found that metabolic impairment correlated with significantly attenuated ATP production(P<0.01)in exper-imental subjects.Moreover,the expression levels of AMPK and PGC-1α mRNA and proteins in lung tissue were signifi-cantly decreased(P<0.01).Moreover,compared to the COPD group,the BYF group showed significant improvements in several of the above indicators,albeit to different degrees(P<0.01 or P<0.05).Moreover,BYF was more effective than NAC in improving minute ventilation and up-regulating PGC-1α expression(P<0.05).CONCLUSION:Bufei-Yishen formula may ameliorate mitochondrial damage in rats with chronic obstructive pulmonary disease by regulating the AMPK/PGC-1α signaling pathway.

AIM:This study aimed to explore the mechanism by which Bufei-Yishen formula(BYF)mitigates mitochondrial damage in rats with chronic obstructive pulmonary disease(COPD)by regulating the AMPK/PGC-1α signal-ing pathway.METHODS:Forty rats were randomly divided into four groups,each containing ten rats each:control group,COPD group,BYF group,and N-acetylcysteine(NAC)group.The COPD model was established through chronic cigarette smoke exposure combined with periodic bacterial inoculations over an eight-week induction phase.During the subsequent eight-week treatment period(i.e.,weeks 9～16),rats in the control and COPD groups received an isovolumet-ric saline solution via oral gavage,at a standardized daily dose of 2 mL per animal.Moreover,rats in the BYF and NAC groups were given Bufei Yishen formula(11.61 g·kg-1·d-1)or N-acetylcysteine(54 mg·kg-1·d-1)by gavage,once per day.At week 16,samples were collected and the general condition of the rats was observed.Body weight was recorded weekly.We also obtained data characterizing rat lung function,lung pathology,ATP content,and mitochondrial ultra-structure,as well as the levels of interleukin-6(IL-6),tumor necrosis factor-alpha(TNF-α),interleukin-1β(IL-1β),se-rum transforming growth factor-beta 1(TGF-β1)and the enzymatic activities of mitochondrial electron transport chain complexes I(NADH dehydrogenase)and III(cytochrome c reductase).Finally,we quantified the mRNA and protein lev-els of AMPK and PGC-1α in lung tissue.RESULTS:Compared to the control group,the COPD group exhibited yellow-ish hair color,reduced gloss,slower weight gain,and a disordered respiratory rhythm.We also observed significant de-creases(P<0.01)in pulmonary function tidal volume(TV),minute ventilation(MV),peak expiratory flow(PEF),expi-ratory flow at 50%of tidal volume(EF50),forced vital capacity(FVC),forced expiratory volume in 0.1 s(FEV0.1),and FEV0.1/FVC.Histopathological analysis showed alveolar cavity enlargement,bullous changes in lung morphology,smooth muscle hypertrophy in the tracheal wall,ciliary destroyed,mucosal shrinking and thickening,and a large number of in-flammatory cells gathered around the tube.Moreover,the mean linear intercept(MLI)and bronchial wall thickness(BWt)had both significantly increased(P<0.01).Electron microscopic analysis of the lungs revealed a reduction in the number of mitochondria in alveolar epithelial cells,a swollen and deformed lung morphology overall.We observed that the mitochondrial cristae were broken,dissolved or vacuolated,accompanied by a significant reduction in the number of lamel-lar bodies and lung volume,along with a disordered internal lipid layer structure.Furthermore,some lung samples were vacuolated or had content leakage.Further quantitative analyses showed statistically significant increases(P<0.01)in the levels of serum pro-inflammatory mediators,including IL-6,TNF-α,IL-1β,and TGF-β1.At the same time we observed substantial reductions in the enzymatic activities of mitochondrial electron transport chain complexes I and III(P<0.01).Moreover,we found that metabolic impairment correlated with significantly attenuated ATP production(P<0.01)in exper-imental subjects.Moreover,the expression levels of AMPK and PGC-1α mRNA and proteins in lung tissue were signifi-cantly decreased(P<0.01).Moreover,compared to the COPD group,the BYF group showed significant improvements in several of the above indicators,albeit to different degrees(P<0.01 or P<0.05).Moreover,BYF was more effective than NAC in improving minute ventilation and up-regulating PGC-1α expression(P<0.05).CONCLUSION:Bufei-Yishen formula may ameliorate mitochondrial damage in rats with chronic obstructive pulmonary disease by regulating the AMPK/PGC-1α signaling pathway.

Objective To evaluate the application value of golden angle radial sparse parallel(Grasp)sequence in contrast-enhanced MRI of liver.Methods The imaging data of 30 patients who underwent gadolinium ethoxybenzyl diethylenetriamine pentaacetic acid contrast-enhanced MRI of liver were collected.With the same equipment,images were collected by Grasp sequence and breath-hold sequence separately,with an interval of less than 3 months.The subjective and objective scores of the late arterial phase and portal venous phase images were evaluated.Results There were no significant differences in all subjective scores of the late arterial phase and portal venous phase images between the two sequences(P＞0.05).The signal-to-noise ratio(SNR)of the late arterial phase images in the Grasp sequence was lower than that in the breath-hold sequence(148.4±52.8 vs 195.6±68.4),and the difference was statistically significant(P＜0.01).Although the SNR of the portal venous phase in the Grasp sequence was lower than that in the breath-hold sequence,the difference was not statistically significant(P＞0.05).There was no statistical difference in the other objective scores between the two sequences(P＞0.05).Conclusion The image quality of Grasp sequence in contrast-enhanced MRI of liver can meet the diagnositic requirements,and it is suitable for patients with poor breath-hold capacity,which has important application value.

Objective To investigate the roles of three Tiao-Bu Fei-Shen Traditional Chinese Medicine(TCM)therapies in improving airway mucus hypersecretion in rats with stable chronic obstructive pulmonary disease(COPD).Methods Ninety rats were divided randomly into nine groups:control(Control)group,model(COPD)group,Bu-Fei Jian-Pi Formula(BJF)group,Bu-Fei Yi-Shen Formula(BYF)group,Yi-Qi Zi-Shen Formula(YZF)group,ERK1/2 inhibitor(PD98059)group,Bu-Fei Jian-Pi combined with inhibitor(BJF+PD98059)group,Bu-Fei Yi-Shen combined with inhibitor(BYF+PD98059)group,and Yi-Qi Zi-Shen combined with inhibitor(YZF+PD98059)group.A rat model of COPD was established by exposing rats to cigarette smoke followed by repeated bacterial infection from weeks 1～8.From weeks 9～16,rats in the control and COPD groups were given 2 mL normal saline,rats in the BJF,BYF,and YZF groups were given the three Tiao-Bu Fei-Shen formulas by gavage,and rats in the PD98059,BJF+PD98059,BYF+PD98059,and YZF+PD98059 groups were given PD98059 by intraperitoneal injection for 7 days at the 16th week.Lung function tests were conducted after 16 weeks and lung tissue morphology,lung water content,inflammatory cell count in bronchoalveolar lavage fluid,and serum levels of inflammatory factors were also assessed.Goblet cell proportion was determined by Alcian blue-periodic acid-Schiff staining,and Muc5AC and Muc5B expression levels were detected by immunohistochemistry.mRNA expression levels of ERK1,ERK2,ENaC,CFTR,and AQP5 were detected by polymerase chain reaction and protein expression levels of ERK1/2 and P-ERK1/2 in lung tissue were determined by Western Blot.Results TV,MV,FVC,FEV0.1,FEV0.1/FVC were significantly decreased(P＜0.01)in COPD rats compared with those in the control group.Lung pathology revealed alveolar disorder,massive fracture of the alveolar wall,and severe shrinkage/thickening of the airway wall accompanied by extensive infiltration of inflammatory cells.Lung tissue water content was significantly increased in COPD rats(P＜0.01),while the proportion of macrophages in BALF was significantly reduced(P＜0.01)and the proportions of neutrophils and lymphocytes were significantly increased(P＜0.01).Serum levels of TNF-α and IL-1β were significantly increased in COPD rats(P＜0.05,P＜0.01).The percentage of goblet cells and expression levels of Muc5AC and Muc5B in airway epithelial cells were significantly increased(P＜0.01),mRNA expression levels of ERK1,ERK2,and ENaC in lung tissue were significantly elevated(P＜0.01),while mRNA expression levels of CFTR and AQP5 were significantly decreased(P＜0.01)in COPD rats compared with levels in the control group.The expression of P-ERK1/2,ERK1/2 in lung tissue was significantly increased(P＜0.01)Rats in the treatment groups demonstrated improvements in the above indicators(P＜0.05,P＜0.01)compared with the COPD group,the groups receiving the three Tiao-Bu Fei-Shen formulas combined with PD98059 showing superior efficacy compared with the single treatment groups(P＜0.05,P＜0.01).Conclusions The three tested Tiao-Bu Fei-Shen therapies can ameliorate airway mucus hypersecretion in COPD rats by inhibiting the ERK1/2 signaling pathway.

Objective:To investigate the relationship between serum soluble CD155 (sCD155), soluble CD163 (sCD163) and chemotherapy efficacy and prognosis in patients with diffuse large B-cell lymphoma (DLBCL).Methods:A total of 126 patients with DLBCL admitted to Handan Central Hospital from May 2018 to May 2020 (DLBCL group) and 126 healthy subjects (control group) were prospectively selected to compare serum sCD155 and sCD163 levels. According to the chemotherapy effect of DLBCL patients, they were divided into effective group and ineffective group, and the serum sCD155 and sCD163 levels were compared before and after treatment. The effective rate of chemotherapy in patients with different serum sCD155 and sCD163 levels was compared. Kaplan-Meier method was used to analyze the relationship between serum sCD155 and sCD163 levels and 3-year overall survival (OS) and progression-free survival (PFS) of DLBCL patients. Cox proportional risk regression model was used to analyze the prognostic factors of DLBCL patients.Results:The serum levels of sCD155 and sCD163 in DLBCL group were higher than those in control group before treatment (all P<0.05). The effective rate of chemotherapy in 126 DLBCL patients in this group was 69.8%(88/126). Compared with the effective group, the serum levels of sCD155 and sCD163 were higher in the ineffective group before and after treatment (all P<0.05). Compared with before treatment, serum sCD155 and sCD163 levels in the effective group were decreased after treatment (all P<0.05). The effective rate of DLBCL patients in sCD155 and sCD163 high level groups was lower than that in sCD155 and sCD163 low level groups (all P<0.05). Kaplan-Meier analysis showed that the 3-year OS and PFS of DLBCL patients in the low level group of sCD155 and sCD163 were higher than those in the high level group (all P<0.05). The high level of sCD155 and sCD163 were independent risk factors for 3-year PFS and OS in DLBCL patients (all P<0.05). Conclusions:Abnormal levels of serum sCD155 and sCD163 in DLBCL patients may reduce the efficacy of chemotherapy and lead to poor prognosis.

Objective To investigate the mechanism by which Xuanfei Jiedu Formula(XFJDF)ameliorates pulmonary damage in rats with multidrug-resistant Pseudomonas aeruginosa(MDR-PA)pneumonia,by modulating the activity of the inhibitor of nuclear factor(NF)-κB kinase(IKK)IKK/NF-κB signal transduction cascade.Methods Eighty-four rats were divided randomly into seven groups:control,model,XFJDF-low dose,XFJDF-medium dose,XFJDF-high dose,imipenem(IPM),and pyrrolidinedithiocarbamate ammonium(PDTC)groups(n=12 rats per group).A MDR-PA pneumonia rat model was established by oral tracheal intubation.After successful model construction,rats in the low-,medium-,and high-dose XFJDF groups were given the corresponding dose of drugs by gavage,rats in the IPM group were given an intraperitoneal injection of IPM,and rats in the control and model groups were given the same volume of normal saline by gavage,twice a day for 7 days.PDTC was injected intraperitoneally 1 h before model establishment,12 h and 24 h after model establishment in the NF-κB inhibitor group.The behavior status,body weight changes,spleen and thymus indexes,and lung wet weight/dry weight ratio were observed in the different groups.Histological changes in the lung tissue were assessed by hematoxylin and eosin staining.Terminal deoxynucleotidyl transferase dUTP nick end labeling was used to detect apoptosis of lung tissue cells,and serum levels of interleukin(IL)-1 β,tumor necrosis factor(TNF)-α,transforming growth factor(TGF)-β,and IL-10 were detected by enzyme-linked immunosorbent assay,and glutathione(GSH)and malondialdehyde(MDA)levels,myeloperoxidase(MPO)activity and total antioxidant capacity(T-AOC)were determined by colorimetry and thiobarbituric acid assay.NF-κBp65 in lung tissue was identified by immunohistochemical analysis,IKKβ and NF-κBp65 mRNA were analyzed by reverse transcription quantitative polymerase chain reaction,and expression levels of IKKβ,phosphorylated(p)-IKKβ,NF-κBp65,and p-NF-κBp65 were measured by Western blot.Results Compared with the control group,rats in the model group exhibited decreased appetite,dull fur,sluggish responsiveness,decreased mobility,increased respiratory rate,audible murmurs,and notable weight loss(P＜0.01).The spleen and thymus indices were significantly enhanced(P＜0.01)and the lung wet/dry weight ratio was significantly increased(P＜0.01),concurrent with increased alveolar secretions in lung tissue.Infiltration of abundant inflammatory cells and increased apoptosis in lung tissue were also noted.Serum concentrations of IL-1β,TNF-α,TGF-β,and IL-10 were increased(P＜0.01)and the MDA content and MPO activity were also increased(P＜0.01).Conversely,GSH levels and T-AOC were significantly decreased(P＜0.01).Lung levels of IKKβ and NF-κBp65 mRNA were significantly increased(P＜0.01)and the ratios of p-IKKβ/IKKβ and p-NF-κBp65/NF-κBp65 were also significantly increased(P＜0.01)compared with the control group.XFJDF,IPM and PDTC improved these parameters to varying degrees,compared with the model group(P＜0.05,P＜0.01),with particularly significant effects in the high-dose XFJDF and IPM groups.Conclusions XFJDF may improve lung injury in rats with MDR-PA pneumonia by inhibiting the IKK/NF-κB signaling pathway.

Objective To investigate the mechanism by which Xuanfei Jiedu Formula(XFJDF)ameliorates pulmonary damage in rats with multidrug-resistant Pseudomonas aeruginosa(MDR-PA)pneumonia,by modulating the activity of the inhibitor of nuclear factor(NF)-κB kinase(IKK)IKK/NF-κB signal transduction cascade.Methods Eighty-four rats were divided randomly into seven groups:control,model,XFJDF-low dose,XFJDF-medium dose,XFJDF-high dose,imipenem(IPM),and pyrrolidinedithiocarbamate ammonium(PDTC)groups(n=12 rats per group).A MDR-PA pneumonia rat model was established by oral tracheal intubation.After successful model construction,rats in the low-,medium-,and high-dose XFJDF groups were given the corresponding dose of drugs by gavage,rats in the IPM group were given an intraperitoneal injection of IPM,and rats in the control and model groups were given the same volume of normal saline by gavage,twice a day for 7 days.PDTC was injected intraperitoneally 1 h before model establishment,12 h and 24 h after model establishment in the NF-κB inhibitor group.The behavior status,body weight changes,spleen and thymus indexes,and lung wet weight/dry weight ratio were observed in the different groups.Histological changes in the lung tissue were assessed by hematoxylin and eosin staining.Terminal deoxynucleotidyl transferase dUTP nick end labeling was used to detect apoptosis of lung tissue cells,and serum levels of interleukin(IL)-1 β,tumor necrosis factor(TNF)-α,transforming growth factor(TGF)-β,and IL-10 were detected by enzyme-linked immunosorbent assay,and glutathione(GSH)and malondialdehyde(MDA)levels,myeloperoxidase(MPO)activity and total antioxidant capacity(T-AOC)were determined by colorimetry and thiobarbituric acid assay.NF-κBp65 in lung tissue was identified by immunohistochemical analysis,IKKβ and NF-κBp65 mRNA were analyzed by reverse transcription quantitative polymerase chain reaction,and expression levels of IKKβ,phosphorylated(p)-IKKβ,NF-κBp65,and p-NF-κBp65 were measured by Western blot.Results Compared with the control group,rats in the model group exhibited decreased appetite,dull fur,sluggish responsiveness,decreased mobility,increased respiratory rate,audible murmurs,and notable weight loss(P＜0.01).The spleen and thymus indices were significantly enhanced(P＜0.01)and the lung wet/dry weight ratio was significantly increased(P＜0.01),concurrent with increased alveolar secretions in lung tissue.Infiltration of abundant inflammatory cells and increased apoptosis in lung tissue were also noted.Serum concentrations of IL-1β,TNF-α,TGF-β,and IL-10 were increased(P＜0.01)and the MDA content and MPO activity were also increased(P＜0.01).Conversely,GSH levels and T-AOC were significantly decreased(P＜0.01).Lung levels of IKKβ and NF-κBp65 mRNA were significantly increased(P＜0.01)and the ratios of p-IKKβ/IKKβ and p-NF-κBp65/NF-κBp65 were also significantly increased(P＜0.01)compared with the control group.XFJDF,IPM and PDTC improved these parameters to varying degrees,compared with the model group(P＜0.05,P＜0.01),with particularly significant effects in the high-dose XFJDF and IPM groups.Conclusions XFJDF may improve lung injury in rats with MDR-PA pneumonia by inhibiting the IKK/NF-κB signaling pathway.

Breast cancer is one of the most serious diseases threatening women's life and health in the world, and the mortality rate is the second in the world. With the progress of nanotechnology and the advantages of nanomaterials in the field of electrochemistry and biosensor, various nanomaterials have been applied in electrochemical biosensors. This makes the electrochemical nano-biosensor in the field of rapid detection of breast cancer has been widely concerned and studied. This paper introduces the important components of electrochemical nano-biosensor for breast cancer detection and the research progress of each component in breast cancer detection, as well as the performance of electrochemical nano biosensor in breast cancer detection and the prospect of its application.
Biosensing Techniques ; Breast Neoplasms/diagnosis* ; Electrochemical Techniques ; Female ; Humans ; Nanostructures ; Nanotechnology

Objective: To investigate differentiation direction of hepatic progenitor cells (HPCs) in cholestatic liver fibrosis (CLF), and the role of Notch signaling pathway in the differentiation of HPCs. Methods: A CLF rat model was established by bile duct ligation (BDL) followed by monitoring changes of Notch signal pathway and the cellular origin of proliferating cholangiocytes. After intraperitoneal injection of DAPT (a Notch signaling inhibitor) after bile duct ligation, the progress of liver fibrosis and the proliferation of cholangiocytes after inhibition of the Notch pathway were analyzed. Results: Data showed that bile duct proliferation gradually increased along with inflammatory cell infiltration and proliferating bile duct cells surrounded by abundant collagen in the BDL group. Immunostaining confirmed markedly increased expression of CK19, OV6, Sox9 and EpCAM. In addition, RT-PCR results showed that Notch signaling pathway was activated significantly. Once the Notch signaling pathway was inhibited by DAPT, bile duct proliferation markedly suppressed along with significantly decreased the mRNA expression of CK19, OV6, Sox9 and EpCAM, compared with BDL group [(10.2±0.7) vs. (22.3±0.8), (7.6±1.5) vs. (18.1±3.7), (1.4±0.4) vs. (4.1±1.1), (1.3±0.3) vs. (5.0±1.4), respectively, P<0.01]. Moreover, liver fibrosis was also reduced significantly. Conclusion Notch signaling activation is required for HPCs differentiation into cholangiocytes in CLF and inhibition of the Notch signaling pathway may offer a therapeutic option for treating CLF.