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.

To explore the chemical constituents of Ecballium elaterium and their cytotoxicity, this study employed multiple chromatographic techniques including normal-phase silica gel, MCI, octadecylsilyl(ODS), Sephadex LH-20 gel, and semi-preparative liquid chromatography for compound isolation from its active fraction. A total of 12 compounds were obtained, and they were identified according to the analysis of a variety of spectral data and literature comparison as 24Z-20,27-dihydroxy-16α,23α-epoxy-cucurbita-2-O-α-L-rhamnopyranosyl-(1→2)-β-D-glucopyranoside(1), cucurbitacin R(2), cucurbitacin B(3), cucurbitacin D(4), cucurbitacin I(5), cucurbitacin L(6), dehydrodiconiferyl alcohol(7), 3-hydroxy-4-methoxycinnamic acid(8), ferulaic acid(9), p-coumaric acid(10), rutin(11), and lariciresinol-4'-O-β-D-glucoside(12), among which compound 1 was a new compound. Compounds 2-6 had strong cytotoxicity against human lung carcinoma A549 cells with the IC_(50) values of(0.48±0.09),(0.03±0.002),(0.13±0.03),(0.87±0.14),(0.15±0.03) μmol·L~(-1), respectively, which were stronger than the positive control doxorubicin \[IC_(50)=(3.92±1.60) μmol·L~(-1)\].
Humans ; Drugs, Chinese Herbal/toxicity* ; Cell Line, Tumor ; Cucurbitaceae/chemistry* ; Cell Survival/drug effects*

Three taraxerane nortriterpenoids were isolated from mastic by using various modern chromatographic separation techniques. They were identified as(5R,8R,9R,10S,11S,12R,13S,17R,18R)-28-norlupa-11,12-epoxy-14-taraxerene-3,16-dione(1),(5R,8R,9R,10S,11S,12R,13S,17S,18S)-17-hydroxy-28-norlupa-11,12-epoxy-14-taraxerene-3-one(2), and(5R,8R,9R,10R,11S,12R,13R,14S,17S,18S)-14,17-epoxy-28-norlupa-11,12-oxidotaraxerone(3) through the high-resolution electrospray ionization mass spectrometry(HR-ESI-MS), infrared(IR), ultraviolet(UV), nuclear magnetic resonance(NMR), and single-crystal X-ray diffraction techniques as well as comparison with literature data. Compounds 1-3 were C-28 nortriterpenoids and isolated from mastic for the first time, and compounds 1-2 were new ones. In the model for RAW264.7 cell anti-inflammation induced by lipopolysaccharide(LPS), compound 1 demonstrates an inhibitory effect on nitric oxide(NO) [IC_(50)=(13.38±0.68) μmol·L~(-1)], comparable to the activity of the positive control dexamethasone [IC_(50)=(14.59±1.49) μmol·L~(-1)]. Compounds 2 and 3 exhibit weaker inhibitory effects, with IC_(50) values of(24.17±2.56) and(22.25±2.84) μmol·L~(-1), respectively.
Animals ; Mice ; Triterpenes/isolation & purification* ; Drugs, Chinese Herbal/isolation & purification* ; Mastic Resin/chemistry* ; Nitric Oxide ; Molecular Structure ; Macrophages/immunology* ; RAW 264.7 Cells

ObjectiveTo investigate the effect and potential mechanism of caffeic acid （CA） on severe acute pancreatitis （SAP） induced by caerulein combined with lipopolysaccharide （LPS）， and to provide a basis for the research on novel drugs for the treatment of SAP. MethodsC57BL/6J mice， aged 6 weeks， were divided into control group， model group， CA group， and octreotide acetate （OA） group， with 6 mice in each group. The mice in the control group were given injection of normal saline， and those in the other groups were given intraperitoneal injection of caerulein combined with LPS to establish a mouse model of SAP. At 1 hour after the first injection of caerulein， the mice in the CA group and the OA group were given intraperitoneal injection of CA or subcutaneous injection of OA at an interval of 8 hours. The general status of the mice was observed after 24 hours of modeling， and serum， pancreas， lung， and colon samples were collected. HE staining was used to observe the histopathological changes of the pancreas and lungs， and the serum levels of α-amylase， lipase， tumor necrosis factor-α （TNF-α）， interleukin-6 （IL-6）， alanine aminotransferase， aspartate aminotransferase， and creatinine were measured. RT-PCR was used to measure the expression of proinflammatory factors in the pancreas and lungs； myeloperoxidase （MPO） immunohistochemistry was used to observe the degree of neutrophil infiltration； Western blot was used to measure the activation of nuclear factor-kappa B （NF-κB） and the level of citrullinated histone H3 （CitH3）， a marker for the formation of neutrophil extracellular traps （NETs）， in the pancreas and lungs， as well as the expression level of ZO-1 in colon tissue. A one-way analysis of variance was used for comparison of continuous data between multiple groups， and the Dunnett’s t-test was used for further comparison between two groups. ResultsCompared with the control group， the model group had severe injury in the pancreas and lungs and significant increases in the activity of serum α- amylase and lipase and the levels of the proinflammatory cytokines IL-6， interleukin-1β （IL-1β）， and TNF-α in serum and lung tissue （all P<0.05）， as well as significant increases in NF-κB activation， neutrophil infiltration， and the formation of NETs in the pancreas and lungs （all P<0.05）. Compared with the model group， the CA group had alleviated pathological injury of the pancreas and lungs and significant reductions in the activity of serum α-amylase and the levels of the proinflammatory cytokines IL-6， IL-1β， and TNF-α in serum and lung tissue （all P<0.05）， as well as significant reductions in NF-κB activation， neutrophil infiltration， and the formation of NETs in the pancreas and lungs （all P<0.05）. ConclusionCA can alleviate SAP induced by caerulein combined with LPS in mice， possibly by inhibiting neutrophil recruitment and the formation of NETs.

Breast cancer (BC) is one of the most prevalent malignant tumors affecting women worldwide, with its incidence rate continuously increasing. As a result, treatment strategies for this disease have received considerable attention. Research has highlighted the crucial role of the Hedgehog (Hh) signaling pathway in the initiation and progression of BC, particularly in promoting tumor growth and metastasis. Therefore, molecular targets within this pathway represent promising opportunities for the development of novel BC therapies. This study aims to elucidate the therapeutic mechanisms by which natural compounds modulate the Hh signaling pathway in BC. By conducting a comprehensive review of various natural compounds, including polyphenols, terpenes, and alkaloids, we reveal both common and unique regulatory mechanisms that influence this pathway. This investigation represents the first comprehensive analysis of five distinct mechanisms through which natural compounds modulate key molecules within the Hh pathway and their impact on the aggressive behaviors of BC. Furthermore, by exploring the structure-activity relationships between these compounds and their molecular targets, we shed light on the specific structural features that enable natural compounds to interact with various components of the Hh pathway. These novel insights contribute to advancing the development and clinical application of natural compound-based therapeutics. Our thorough review not only lays the groundwork for exploring innovative BC treatments but also opens new avenues for leveraging natural compounds in cancer therapy.

Objective:To analyze the risk factors for growth disturbance (GD) in children with distal femoral epiphyseal fracture (DFEF) after surgical treatment.Methods:A retrospective study was conducted to analyze the clinical data of the 72 children who had undergone surgery for DFEF at Department of Pediatric Orthopaedics, The Second General Hospital of Fuzhou between February 2013 and February 2024. There were 52 boys and 20 girls with an age of 11.0 (5.0, 13.0) years. The data collected included age at injury, gender, side affected, cause for injury, time from injury to surgery, the maximum fracture displacement, Salter-Harris fracture classification, and presence of high-energy trauma. The risk factors for GD after DFEF surgical treatment were determined through univariate analysis and logistic regression analysis.Results:Distal femur GD occurred in 40.2% (29/72) of the children treated surgically for DFEF. The univariate analysis showed that, compared with the children without GD, those with GD had a significantly significantly longer time from injury to surgery ( P=0.005), a significantly greater fracture displacement ( P=0.002), and more severe Salter-Harris fracture classification ( P=0.045). The logistic analysis showed that all the 3 factors were independent risk factors for GD ( P<0.05). Conclusion:After DFEF surgery, the GD risk is significantly increased by the 3 factors:longer time from injury to surgery, greater fracture displacement, and more severe fracture classification.

Breast cancer(BC)is one of the most prevalent malignant tumors affecting women worldwide,with its incidence rate continuously increasing.As a result,treatment strategies for this disease have received considerable attention.Research has highlighted the crucial role of the Hedgehog(Hh)signaling pathway in the initiation and progression of BC,particularly in promoting tumor growth and metastasis.There-fore,molecular targets within this pathway represent promising opportunities for the development of novel BC therapies.This study aims to elucidate the therapeutic mechanisms by which natural com-pounds modulate the Hh signaling pathway in BC.By conducting a comprehensive review of various natural compounds,including polyphenols,terpenes,and alkaloids,we reveal both common and unique regulatory mechanisms that influence this pathway.This investigation represents the first comprehen-sive analysis of five distinct mechanisms through which natural compounds modulate key molecules within the Hh pathway and their impact on the aggressive behaviors of BC.Furthermore,by exploring the structure-activity relationships between these compounds and their molecular targets,we shed light on the specific structural features that enable natural compounds to interact with various components of the Hh pathway.These novel insights contribute to advancing the development and clinical application of natural compound-based therapeutics.Our thorough review not only lays the groundwork for exploring innovative BC treatments but also opens new avenues for leveraging natural compounds in cancer therapy.

To investigate the immune protective effect induced by superoxide dismutase(SOD)of the Toxocara canis(T.canis)in mice,in this experiment,seventy-five Kunming mice of six-week-age were randomly divided into 5 groups for the immune protection experiment,the mice were subcutaneously immunized with 50,75 and 125 mg/L Tc-SOD(0.1 mL)on days 0,14 and 28,re-spectively.In the blank group,mice did not receive any treatment,while the PBS group was injected with equal amounts of sterility PBS.A total of three immunizations and each immunization interval of 14 days.14 days after the third immunization,the 3 000 infected worm eggs were given orally in all groups.Before each immunization,14 days after the third immunization,and after the T.canis attack on the 7th day,the blood was sampled from the tail vein and sera were separated,while the IgG levels in serum were detected by indirect ELISA.The proliferation ability of splenic lympho-cytes and the mRNA expression levels of Th1(IFN-γ,IL-2)and Th2 cytokines(IL-4,IL-10)were analyzed with CCK-8 and qRT-PCR.The larvae reduction rates were calculated on the 7th day after the T.canis attack,and the pathological changes in the livers and lungs were observed using H&E staining.The results showed that compared with the PBS group,the serum IgG antibody levels increased with the frequency and duration of Tc-SOD immunization.It remained at a high level af-ter the T.canis attack,and the differences were very significant(P＜0.001).The mRNA expres-sion levels of IFN-γ,IL-2,IL-4,and IL-10 were dramatically increased(P＜0.01),while IL-4 andIL-10 were significantly higher than IFN-γ and IL-2 on the 14th day after the third immunization(P＜0.05),showing the Th2 type predominant immune response.And induce the proliferation of spleen lymphocytes in vitro(P＜0.01).The reduction rates of larvae of 50,75 and 125 mg/L Tc-SOD immune groups were 18.7％,24.9％and 37.6％,respectively,with significant differences in the 125 mg/L Tc-SOD group(P＜0.05),and had better immune protection effect.Moreover,the H&E staining results indicated that three Tc-SOD immune groups reduce the inflammatory cell infiltration(neutrophils,eosinophils,and macrophages)and hemorrhagic lesions in the livers and lungs of mice.The above results indicated that Tc-SOD could induce humoral and cellular immune responses in mice,mainly Th2 immune response,and provide immune protection against T.canis infection.

To investigate the immune protective effect induced by superoxide dismutase(SOD)of the Toxocara canis(T.canis)in mice,in this experiment,seventy-five Kunming mice of six-week-age were randomly divided into 5 groups for the immune protection experiment,the mice were subcutaneously immunized with 50,75 and 125 mg/L Tc-SOD(0.1 mL)on days 0,14 and 28,re-spectively.In the blank group,mice did not receive any treatment,while the PBS group was injected with equal amounts of sterility PBS.A total of three immunizations and each immunization interval of 14 days.14 days after the third immunization,the 3 000 infected worm eggs were given orally in all groups.Before each immunization,14 days after the third immunization,and after the T.canis attack on the 7th day,the blood was sampled from the tail vein and sera were separated,while the IgG levels in serum were detected by indirect ELISA.The proliferation ability of splenic lympho-cytes and the mRNA expression levels of Th1(IFN-γ,IL-2)and Th2 cytokines(IL-4,IL-10)were analyzed with CCK-8 and qRT-PCR.The larvae reduction rates were calculated on the 7th day after the T.canis attack,and the pathological changes in the livers and lungs were observed using H&E staining.The results showed that compared with the PBS group,the serum IgG antibody levels increased with the frequency and duration of Tc-SOD immunization.It remained at a high level af-ter the T.canis attack,and the differences were very significant(P＜0.001).The mRNA expres-sion levels of IFN-γ,IL-2,IL-4,and IL-10 were dramatically increased(P＜0.01),while IL-4 andIL-10 were significantly higher than IFN-γ and IL-2 on the 14th day after the third immunization(P＜0.05),showing the Th2 type predominant immune response.And induce the proliferation of spleen lymphocytes in vitro(P＜0.01).The reduction rates of larvae of 50,75 and 125 mg/L Tc-SOD immune groups were 18.7％,24.9％and 37.6％,respectively,with significant differences in the 125 mg/L Tc-SOD group(P＜0.05),and had better immune protection effect.Moreover,the H&E staining results indicated that three Tc-SOD immune groups reduce the inflammatory cell infiltration(neutrophils,eosinophils,and macrophages)and hemorrhagic lesions in the livers and lungs of mice.The above results indicated that Tc-SOD could induce humoral and cellular immune responses in mice,mainly Th2 immune response,and provide immune protection against T.canis infection.

Objective:To analyze the risk factors for growth disturbance (GD) in children with distal femoral epiphyseal fracture (DFEF) after surgical treatment.Methods:A retrospective study was conducted to analyze the clinical data of the 72 children who had undergone surgery for DFEF at Department of Pediatric Orthopaedics, The Second General Hospital of Fuzhou between February 2013 and February 2024. There were 52 boys and 20 girls with an age of 11.0 (5.0, 13.0) years. The data collected included age at injury, gender, side affected, cause for injury, time from injury to surgery, the maximum fracture displacement, Salter-Harris fracture classification, and presence of high-energy trauma. The risk factors for GD after DFEF surgical treatment were determined through univariate analysis and logistic regression analysis.Results:Distal femur GD occurred in 40.2% (29/72) of the children treated surgically for DFEF. The univariate analysis showed that, compared with the children without GD, those with GD had a significantly significantly longer time from injury to surgery ( P=0.005), a significantly greater fracture displacement ( P=0.002), and more severe Salter-Harris fracture classification ( P=0.045). The logistic analysis showed that all the 3 factors were independent risk factors for GD ( P<0.05). Conclusion:After DFEF surgery, the GD risk is significantly increased by the 3 factors:longer time from injury to surgery, greater fracture displacement, and more severe fracture classification.