ObjectiveTo investigate the therapeutic effect of wall-broken ganoderma lucidum spore powder （GLS） on a mouse model of hepatocellular carcinoma and to explore its underlying mechanism. MethodsFifty 2-week-old C57BL/6J mice were divided into a control group， a model group and low， medium and high-dose GLS groups（0.25， 0.5， 1.0 g/kg ）， with 10 mice in each group. A DEN/CCL₄ mouse hepatocellular carcinoma model was established， and GLS was administered by gavage starting from the 8th week， once a day for 14 weeks. At the end of the final administration， the number of liver nodules was observed ， and the serum alpha-fetoprotein （AFP） was detected using enzyme-linked immunosorbent assay. The content of catalase （CAT） in serum and liver tissue was determined by ammonium molybdate method. The content of reduced glutathione （GSH） in serum and liver tissue was measured by microplate assay. The content of glutathione peroxidase （GSH-PX） in serum and liver tissue was detected by colorimetric method. Immunohistochemistry （IHC-P） was employed to detect the protein expression of proliferating cell nuclear antigen （PCNA）. R eal-time fluorescence quantitative polymerase chain reaction was utilized to detect the relative expression of nuclear factor E2-related factor 2 （Nrf2）， Kelch-like epichlorohydrin-associated protein 1 （Keap1）， NAD（P）H quinone oxidoreductase 1 （Nqo1）， heme oxygenase-1 （Ho-1） and glutamate cysteine ligase catalytic subunit （Gclc） mRNA. ResultsCompared with the control group， the content of serum AFP were significantly higher in the model group （P<0.01）， while the contents of CAT， GSH and GSH-PX in liver and serum were significantly lower in the model group （P<0.01）. The relative expression of Keap1 and Nqo1 mRNA was significantly higher （P<0.01）， the relative expression of Nrf2， Ho-1， Gclc mRNA relative expression was significantly lower （P<0.01）， and the positive expression rate of PCNA was significantly higher （P<0.01）. Compared with the model group， the total number of liver nodules and content of serum AFP in the administered group were significantly lower （P<0.01）， and the contents of CAT， GSH， and GSH-PX in the liver and serum of mice significantly increased （P<0.05 or P<0.01）. The expression of Keap1 and Nqo1 mRNA significantly decreased （P<0.05）. The expression of Nrf2， Ho-1 and Gclc mRNA significantly increased （P<0.05）， and positive expression rate of PCNA significantly decreased （P<0.01）. ConclusionGLS has a significant protective effect on DEN/CCL4 -induced mouse hepatocellular carcinoma model， and the mechanism of action is related to the regulation of oxidative stress in mouse liver.

AIM To study the chemical constituents from ethyl acetate fraction of Balanophora harlandii Hook.f.and their tyrosinase inhibitory activity.METHODS Separation and purification were performed using silica gel,MCI,ODS,Sephadex LH-20 and semi-preparative HPLC,then the structures of obtained compounds were identified by physicochemical properties and spectral data.The monophenolase inhibitory activity was determined by the tyrosinase-catalyzed oxidation of L-tyrosine.RESULTS Twenty-four compounds were isolated and identified as sesamin(1),methyl caffeate(2),quercetin(3),5,7-dihydroxychromanone(4),methyl 3,4-dihydroxybenzoate(5),esculetin(6),kaempferol(7),naringenin(8),pyrogallic acid(9),pinosylvin(10),methyl propionate(11),caffeic acid(12),saccharinol(13),ferulic acid(14),trans-p-hydroxycinnamic acid(15),cinnamic acid(16),vanillic acid(17),vanillin(18),4-hydroxyacetophenone(19),4-hydroxybenzaldehyde(20),apigenin(21),(-)-isolariciresinol(22),(-)-secoisolariciresinol(23)and meso-2,3-di(3′,4′-methylenedioxybenzyl)butane-1,4-diol(24).The IC50 values of compounds 3,5,7,8,19,and 20 ranged from(0.246 5±0.028 3)to(1.278 2±0.021 3)mmol/L.CONCLUSION Compounds 1-9、11、15、17-21、24 are isolated from this plant for the first time,and 1,6,9,17-19,24 are first isolated from genus Balanophora.Compounds 3、5、7、8、19 and 20 have tyrosinase inhibitory activity.

AIM:To investigate the effects of ethylisopropylamiloride(EIPA),an inhibitor of Na+/H+ex-changer 1(NHE1),on the proliferation,migration,invasion,cell cycle and apoptosis of hepatocellular carcinoma cells,and to explore its mechanisms.METHODS:The human hepatocellular carcinoma cell line Huh7 was cultured in vitro,and treated with different concentrations of EIPA.The mRNA and protein expression of NHE1 in Huh7 cells was assessed by RT-qPCR and Western blot.The proliferation of Huh7 cells was examined by kFluor488-EdU staining.The migration and invasion of Huh7 cells were assessed by Transwell assays.The cell cycle distribution and apoptosis of these cells were detected by flow cytometry.Moreover,the intracellular pH was measured via the PTI high-speed ion imaging system.RE-SULTS:Treatment with EIPA significantly suppressed the mRNA and protein expression of NHE1(P＜0.01).It signifi-cantly inhibited the proliferation,migration and invasion of Huh7 cells(P＜0.05 or P＜0.01),promoted their apoptosis(P＜0.01),and caused significant S-phase arrest(P＜0.01).It also significantly reduced the intracellular pH of Huh7 cells.CONCLUSION:The NHE1 inhibitor EIPA regulates the intracellular pH by inhibiting the expression and function of NHE1,which consequently affects the proliferation,migration,invasion,cell cycle,and apoptosis of Huh7 cells.There-fore,NHE1 may be a potential target for the treatment of hepatocellular carcinoma.

AIM To study the chemical constituents from ethyl acetate fraction of Balanophora harlandii Hook.f.and their tyrosinase inhibitory activity.METHODS Separation and purification were performed using silica gel,MCI,ODS,Sephadex LH-20 and semi-preparative HPLC,then the structures of obtained compounds were identified by physicochemical properties and spectral data.The monophenolase inhibitory activity was determined by the tyrosinase-catalyzed oxidation of L-tyrosine.RESULTS Twenty-four compounds were isolated and identified as sesamin(1),methyl caffeate(2),quercetin(3),5,7-dihydroxychromanone(4),methyl 3,4-dihydroxybenzoate(5),esculetin(6),kaempferol(7),naringenin(8),pyrogallic acid(9),pinosylvin(10),methyl propionate(11),caffeic acid(12),saccharinol(13),ferulic acid(14),trans-p-hydroxycinnamic acid(15),cinnamic acid(16),vanillic acid(17),vanillin(18),4-hydroxyacetophenone(19),4-hydroxybenzaldehyde(20),apigenin(21),(-)-isolariciresinol(22),(-)-secoisolariciresinol(23)and meso-2,3-di(3′,4′-methylenedioxybenzyl)butane-1,4-diol(24).The IC50 values of compounds 3,5,7,8,19,and 20 ranged from(0.246 5±0.028 3)to(1.278 2±0.021 3)mmol/L.CONCLUSION Compounds 1-9、11、15、17-21、24 are isolated from this plant for the first time,and 1,6,9,17-19,24 are first isolated from genus Balanophora.Compounds 3、5、7、8、19 and 20 have tyrosinase inhibitory activity.

High-performance liquid chromatography(HPLC) was used to determine the content of three major components in Citri Reticulatae Semen(CRS), including limonin, nomilin, and obacunone. The chromaticity of the CRS sample during salt processing and stir-frying was measured using a color difference meter. Next, the relationship between the color and content of the salt-processed CRS sample was investigated through correlation analysis. By integrating the oil bath technique for processing simulation with HPLC, the changes in the relative content of nomilin and its transformation products were analyzed, with its structural transformation pattern during processing identified. Additionally, RAW264.7 cells were induced with lipopolysaccharides(LPSs) to establish an inflammatory model, and the anti-inflammatory activity of nomilin and its transformation product, namely obacunone was evaluated. The results indicated that as processing progressed, E~*ab and L~* values showed a downward trend; a~* values exhibited a slow increase over a certain period, followed by no significant changes, and b~* values remained stable with no significant changes over a certain period and then started to decrease. The limonin content remained barely unchanged; the nomilin content decreased, and the obacunone increased significantly. The changing trends in content and color parameters during salt-processing and stir-frying were basically consistent. The content of nomilin and obacunone was significantly correlated with the colorimetric values(L~*, a~*, b~*, and E~*ab), while limonin content showed no significant correlation with these values. By analyzing HPLC patterns of nomylin at different heating temperatures and time, it was found that under conditions of 200-250 ℃ for heating of 5-60 min, the content of nomilin significantly decreased, while the obacunone content increased pronouncedly. The in vitro anti-inflammatory activity results indicated that compared to the model group, the group with a high concentration of nomilin and the groups with varying concentrations of obacunone showed significantly reduced release of nitric oxide(NO)(P<0.01). When both were at the same concentration, obacunone showed better performance in inhibiting NO release. In this study, the obvious correlation between the color and content of major components during the processing of CRS samples was identified, and the dynamic patterns of quality change in CRS samples during processing were revealed. Additionally, the study revealed and confirmed the transformation of nomilin into obacunone during processing, with the in vitro anti-inflammatory activity of obacunone significantly greater than that of nomilin. These findings provided a scientific basis for CRS processing optimization, tablet quality control, and its clinical application.
Mice ; Animals ; Drugs, Chinese Herbal/pharmacology* ; RAW 264.7 Cells ; Limonins/chemistry* ; Chromatography, High Pressure Liquid ; Citrus/chemistry* ; Color ; Benzoxepins/chemistry* ; Anti-Inflammatory Agents/chemistry*

Objective To establish a rapid,sensitive,and specific multiplex PCR detection method for the simultaneous detection of Cryptosporidium,Eimeria,and bovine parvovirus.Methods Specific primers targeting the SSU rRNA genes of Cryptosporidium and Eimeria,as well as the VP2 gene of bovine parvovirus were designed and the corresponding recombinant plasmid standards were constructed.To establish the multiplex PCR method,the reaction conditions were optimized using temperature gradient PCR and single-variable control methods.The sensitivity,specificity,reproducibility,and clinical application of the protocol were evaluated.Results The optimal annealing temperature was found to be 60.5℃,and the forward and reverse primer concentrations were determined to be 0.2 μmol/L for Eimeria,and 0.4 μmol/L for Cryptosporidium and bovine parvovirus.The assay demonstrated high sensitivity,with detection limits of 243,260,and 3 110 copies for the recombinant plasmid standards of Cryptosporidium,Eimeria,and bovine parvovirus,respectively.Specificity testing showed no cross-reactivity with ten common bovine pathogens,including Salmonella,bovine viral diarrhea virus,and bovine rotavirus.Consistent intra-and inter-batch results confirmed the strong reproducibility of the method.Clinical application to 81 diarrhea samples from various regions in the Ganzi Prefecture,Sichuan,revealed positivity rates of 18.52%(15/81)for Cryptosporidium,34.57%(28/81)for Eimeria,and 18.52%(15/81)forbovineparvovirus,withamixedinfectionrateof3.7%(3/81).Conclusions Themultiplex PCR method established in this study offers a reliable tool for differential diagnosis and epidemiological investigation of the three common diarrheal pathogens in yaks.

OBJECTIVE: Humans are exposed to complex mixtures of environmental chemicals and other factors that can affect their health. Analysis of these mixture exposures presents several key challenges for environmental epidemiology and risk assessment, including high dimensionality, correlated exposure, and subtle individual effects. METHODS: We proposed a novel statistical approach, the generalized functional linear model (GFLM), to analyze the health effects of exposure mixtures. GFLM treats the effect of mixture exposures as a smooth function by reordering exposures based on specific mechanisms and capturing internal correlations to provide a meaningful estimation and interpretation. The robustness and efficiency was evaluated under various scenarios through extensive simulation studies. RESULTS: We applied the GFLM to two datasets from the National Health and Nutrition Examination Survey (NHANES). In the first application, we examined the effects of 37 nutrients on BMI (2011-2016 cycles). The GFLM identified a significant mixture effect, with fiber and fat emerging as the nutrients with the greatest negative and positive effects on BMI, respectively. For the second application, we investigated the association between four pre- and perfluoroalkyl substances (PFAS) and gout risk (2007-2018 cycles). Unlike traditional methods, the GFLM indicated no significant association, demonstrating its robustness to multicollinearity. CONCLUSION GFLM framework is a powerful tool for mixture exposure analysis, offering improved handling of correlated exposures and interpretable results. It demonstrates robust performance across various scenarios and real-world applications, advancing our understanding of complex environmental exposures and their health impacts on environmental epidemiology and toxicology.
Humans ; Environmental Exposure/analysis* ; Linear Models ; Nutrition Surveys ; Environmental Pollutants ; Body Mass Index

Objective To investigate the influence of different disinfectants on the ultrasound-guided internal jugular vein puncture and adhesive application.Methods A total of 102 emergency patients with traumatic hemorrhage who underwent ultrasound-guided internal jugular vein puncture and catheterization were selected as the study subjects and randomly divided into the iodine group(51 cases)and the chlorhexidine-alcohol group(51 cases).Iodophor(0.5%effective iodine)and 2.0%chlorhexidine gluconate+70.0%ethanol were used for disinfection before puncture,respectively.The disinfection effect of the puncture site,puncture condition,ultrasound imaging,and adhesive force of the application of patients were statistically compared between the two groups.Results There was no significant difference in the bacterial count before disinfection,the bacterial count after disinfection or the qualified rate of disinfection at the puncture site of patients between the two groups(P>0.05).There were significant differences in the success rate of one-time puncture,the number of puncture and the time of puncture and catheterization between the two groups(P<0.05).There were significant differences in the rate of clear ultrasound images,adhesive force and rate of warping or shedding of the application between the two groups(P<0.05).Conclusion For emergency patients with traumatic hemorrhage,using iodophor(0.5%effective iodine)and 2.0%chlorhexidine gluconate+70.0%ethanol disinfection before ultrasound-guided internal jugular vein puncture can achieve good disinfection effects.Iodine disinfection can obtain clearer ultrasound images and a higher success rate of one-time puncture,but the adhesive force of the application is poor,making it prone to warping or shedding.

AIM:To investigate the effects of ethylisopropylamiloride(EIPA),an inhibitor of Na+/H+ex-changer 1(NHE1),on the proliferation,migration,invasion,cell cycle and apoptosis of hepatocellular carcinoma cells,and to explore its mechanisms.METHODS:The human hepatocellular carcinoma cell line Huh7 was cultured in vitro,and treated with different concentrations of EIPA.The mRNA and protein expression of NHE1 in Huh7 cells was assessed by RT-qPCR and Western blot.The proliferation of Huh7 cells was examined by kFluor488-EdU staining.The migration and invasion of Huh7 cells were assessed by Transwell assays.The cell cycle distribution and apoptosis of these cells were detected by flow cytometry.Moreover,the intracellular pH was measured via the PTI high-speed ion imaging system.RE-SULTS:Treatment with EIPA significantly suppressed the mRNA and protein expression of NHE1(P＜0.01).It signifi-cantly inhibited the proliferation,migration and invasion of Huh7 cells(P＜0.05 or P＜0.01),promoted their apoptosis(P＜0.01),and caused significant S-phase arrest(P＜0.01).It also significantly reduced the intracellular pH of Huh7 cells.CONCLUSION:The NHE1 inhibitor EIPA regulates the intracellular pH by inhibiting the expression and function of NHE1,which consequently affects the proliferation,migration,invasion,cell cycle,and apoptosis of Huh7 cells.There-fore,NHE1 may be a potential target for the treatment of hepatocellular carcinoma.

AIM To investigate the differences between boiled powder and decoction of Ermiao Powder.METHODS GC-MS was used to identify volatile constituents,after which the content determination of linalool,4-terpineol,α-terpineol,β-eucalyptol and taxifolin in distillate was performed,evaporation rate curve was drawn.The rat model for collagen-induced arthritis was established,then HE and SO/FG staining were conducted,and body weight,footpad swelling degree,arthritis score,immune organ(spleen,thymus)indices,serum inflammatory factors(IL-10,IL-6),ankle joint structure(foot claw swelling,micro-CT)and locomotor ability were detected.RESULTS Total 43 and 26 volatile constituents were identified in boiled powder and decoction,respectively.With the extension of boiling time,the average evaporation rates of 5 volatile constituents in the boiled powder distillate demonstrated the trends of first increase and then decrease,which reached a maximum at 15-20 min;those in the decoction distillate displayed the trends of decrease,which were not detected after 15 min except for β-eucalyptol.Compared with the decoction,the boiled powder exhibited stronger effects on improving foot swelling and arthritis score,alleviating pathological changes in joint tissues,inhibiting inflammatory factors and restoring motor ability.CONCLUSION More volatile constituents are observable in the boiled powder of Ermiao Powder than those in its decoction,along with stronger anti-rheumatoid arthritis activity.The optimal decocting endpoint is determined to be within 15 min in boiling water.