BACKGROUND:Studies have confirmed that Pterocarya hupehensis skan total flavonoids(PHSTF)can improve the level of collagen-induced arthritis in rats,but there is still a lack of research on the regulation of Wnt/β-catenin signaling pathway in fibroblast-like synoviocytes and its effect on related cell functions.OBJECTIVE:To investigate the effect and mechanism of PHSTF on lipopolysaccharide-induced proliferation,migration and apoptosis of fibroblast-like synoviocytes based on the Wnt/β-catenin signaling pathwayMETHODS:Fibroblast-like synoviocytes were divided into control group,lipopolysaccharide group,lipopolysaccharide+low-,medium-,and high-dose PHSTF groups(10,20,and 40 μg/mL),lipopolysaccharide+Wnt pathway inhibitor DKK1 group,and lipopolysaccharide+Wnt pathway inhibitor DKK1+high-dose PHSTF group(40 μg/mL).The cell counting kit-8 method was used to detect the effect of PHSTF on the viability of fibroblast-like synoviocytes,and the final drug concentration and time were screened.Flow cytometry was used to detect the apoptosis of fibroblast-like synoviocytes.Cell scratch assay,EDU staining and cell cloning assay were used to detect the migration and proliferation of fibroblast-like synoviocytes.Western blot assay was used to detect the protein expression levels of Wnt3a,β-catenin,tumorigenic genes,matrix metalloproteinase 2,matrix metalloproteinase 9,Bax and Bcl-2 in fibroblast-like synoviocytes.RESULTS AND CONCLUSION:(1)Compared with the control group,the cell viability decreased significantly when the concentration of PHSTF was＞40 μg/mL(P＜0.01).Therefore,the drug concentration of≤40 μg/mL was selected for subsequent experiments.(2)Compared with the lipopolysaccharide group,the wound healing rate,cell clone formation rate and the number of EDU-positive cells in the low-,medium-and high-dose PHSTF groups were significantly reduced,while the apoptosis rate was significantly increased(P＜0.05-0.01).(3)Western blot results showed that compared with the lipopolysaccharide group,low-,medium-and high-dose PHSTF significantly inhibited cellular Wnt3a,β-catenin,cellular tumorigenic genes,matrix metalloproteinase 2,matrix metalloproteinase 9,and Bcl-2 protein expression,and promoted the expression of Bax protein(P＜0.01).(4)Compared with the DKK1 group,the combination of DKK1 and high-dose PHSTF significantly inhibited the protein expression of Wnt3a,β-catenin,matrix metalloproteinase 2,matrix metalloproteinase 9 and Bcl-2 protein expression and promoted the protein expression of Bax(P＜0.01).To conclude,PHSTF may inhibit the proliferation and migration of fibroblast-like synoviocytes and promote apoptosis by inhibiting the Wnt/β-catenin signaling pathway.

BACKGROUND:Studies have confirmed that Pterocarya hupehensis skan total flavonoids(PHSTF)can improve the level of collagen-induced arthritis in rats,but there is still a lack of research on the regulation of Wnt/β-catenin signaling pathway in fibroblast-like synoviocytes and its effect on related cell functions.OBJECTIVE:To investigate the effect and mechanism of PHSTF on lipopolysaccharide-induced proliferation,migration and apoptosis of fibroblast-like synoviocytes based on the Wnt/β-catenin signaling pathwayMETHODS:Fibroblast-like synoviocytes were divided into control group,lipopolysaccharide group,lipopolysaccharide+low-,medium-,and high-dose PHSTF groups(10,20,and 40 μg/mL),lipopolysaccharide+Wnt pathway inhibitor DKK1 group,and lipopolysaccharide+Wnt pathway inhibitor DKK1+high-dose PHSTF group(40 μg/mL).The cell counting kit-8 method was used to detect the effect of PHSTF on the viability of fibroblast-like synoviocytes,and the final drug concentration and time were screened.Flow cytometry was used to detect the apoptosis of fibroblast-like synoviocytes.Cell scratch assay,EDU staining and cell cloning assay were used to detect the migration and proliferation of fibroblast-like synoviocytes.Western blot assay was used to detect the protein expression levels of Wnt3a,β-catenin,tumorigenic genes,matrix metalloproteinase 2,matrix metalloproteinase 9,Bax and Bcl-2 in fibroblast-like synoviocytes.RESULTS AND CONCLUSION:(1)Compared with the control group,the cell viability decreased significantly when the concentration of PHSTF was＞40 μg/mL(P＜0.01).Therefore,the drug concentration of≤40 μg/mL was selected for subsequent experiments.(2)Compared with the lipopolysaccharide group,the wound healing rate,cell clone formation rate and the number of EDU-positive cells in the low-,medium-and high-dose PHSTF groups were significantly reduced,while the apoptosis rate was significantly increased(P＜0.05-0.01).(3)Western blot results showed that compared with the lipopolysaccharide group,low-,medium-and high-dose PHSTF significantly inhibited cellular Wnt3a,β-catenin,cellular tumorigenic genes,matrix metalloproteinase 2,matrix metalloproteinase 9,and Bcl-2 protein expression,and promoted the expression of Bax protein(P＜0.01).(4)Compared with the DKK1 group,the combination of DKK1 and high-dose PHSTF significantly inhibited the protein expression of Wnt3a,β-catenin,matrix metalloproteinase 2,matrix metalloproteinase 9 and Bcl-2 protein expression and promoted the protein expression of Bax(P＜0.01).To conclude,PHSTF may inhibit the proliferation and migration of fibroblast-like synoviocytes and promote apoptosis by inhibiting the Wnt/β-catenin signaling pathway.

ObjectiveTo establish a model that can quickly identify the aroma components in different parts of Nardostachys jatamansi， so as to provide a quality control basis for the market circulation and clinical use of N. jatamansi. MethodsHeadspace solid-phase microextraction-gas chromatography-mass spectrometry（HS-SPME-GC-MS） combined with Smart aroma database and National Institute of Standards and Technology（NIST） database were used to characterize the aroma components in different parts of N. jatamansi， and the aroma components were quantified according to relative response factor（RRF） and three internal standards， and the markers of aroma differences in different parts of N. jatamansi were identified by orthogonal partial least squares-discriminant analysis（OPLS-DA） and cluster thermal analysis based on variable importance in the projection（VIP） value >1 and P<0.01. The odor data of different parts of N. jatamansi were collected by Heracles Ⅱ Neo ultra-fast gas phase electronic nose， and the correlation between compound types of aroma components collected by the ultra-fast gas phase electronic nose and the detection results of HS-SPME-GC-MS was investigated by drawing odor fingerprints and odor response radargrams. Chromatographic peak information with distinguishing ability≥0.700 and peak area≥200 was selected as sensor data， and the rapid identification model of different parts of N. jatamansi was established by principal component analysis（PCA）， discriminant factor alysis（DFA）， soft independent modeling of class analogies（SIMCA） and statistical quality control analysis（SQCA）. ResultsThe HS-SPME-GC-MS results showed that there were 28 common components in the underground and aboveground parts of N. jatamansi， of which 22 could be quantified and 12 significantly different components were screened out. Among these 12 components， the contents of five components（ethyl isovalerate， 2-pentylfuran， benzyl alcohol， nonanal and glacial acetic acid，） in the aboveground part of N. jatamansi were significantly higher than those in the underground part（P<0.01）， the contents of β-ionone， patchouli alcohol， α-caryophyllene， linalyl butyrate， valencene， 1，8-cineole and p-cymene in the underground part of N. jatamansi were significantly higher than those in the aboveground part（P<0.01）. Heracles Ⅱ Neo electronic nose results showed that the PCA discrimination index of the underground and aboveground parts of N. jatamansi was 82， and the contribution rates of the principal component factors were 99.94% and 99.89% when 2 and 3 principal components were extracted， respectively. The contribution rate of the discriminant factor 1 of the DFA model constructed on the basis of PCA was 100%， the validation score of the SIMCA model for discrimination of the two parts was 99， and SQCA could clearly distinguish different parts of N. jatamansi. ConclusionHS-SPME-GC-MS can clarify the differential markers of underground and aboveground parts of N. jatamansi. The four analytical models provided by Heracles Ⅱ Neo electronic nose（PCA， DFA， SIMCA and SQCA） can realize the rapid identification of different parts of N. jatamansi. Combining the two results， it is speculated that terpenes and carboxylic acids may be the main factors contributing to the difference in aroma between the underground and aboveground parts of N. jatamansi.

Objective To investigate the effect of Qianggu Kangshu Formula(QGKSF)for alleviating osteoclast differentiation in rheumatoid arthritis and the underlying mechanism.Methods RAW264.7 cells cultured under hypoxic conditions were treated with RANKL to induce osteoclast differentiation and incubated with normal rat serum or sera from rats medicated with methotrexate(MTX)or QGKSF at low and high doses.Cell viability,TRAP-positive multinucleated cells and F-actin ring formation in the treated cells were assessed with CCK-8 assay,TRAP staining and Phalloidin staining,respectively.Autophagy and autophagosomes in the cells were observed with MDC staining and transmission electron microscopy.ELISA was used to measure IL-6 and TNF-α levels in the culture supernatant,and the expressions of HIF-1α,BNIP3,Bcl-2,Beclin1,LC3-I,LC3-II,P62 and TRAP mRNAs and proteins were analyzed using RT-qPCR and Western blotting.Results In hypoxia-and RANKL-induced RAW264.7 cells treated with normal rat serum,significant increments of TRAP-positive cells and F-actin ring formation were observed with an enhanced autophagic fluorescence intensity and increased autophagosomes.Treatment of the induced cells with rat sera medicated with MTX and low-and high-dose QGKSF obviously reduced the TRAP-positive cells,F-actin rings and autophagosomes as well as the autophagic fluorescence intensity.RANKL treatment significantly increased IL-6 and TNF-α levels in RAW264.7 cells,which were obviously decreased by treatment with MTX-and QGKSF-medicated sera.RANKL also significantly increased the mRNA and protein expression levels of HIF-1α,BNIP3,Bcl-2,Beclin1,LC3 and TRAP and lowered P62 expressions,and these changes were effectively reversed by treatment with MTX-and QGKSF-medicated sera.Conclusion QGKSF attenuates RANKL-induced osteoclast differentiation in hypoxic RAW264.7 cells by inhibiting the HIF-1α/BNIP3 autophagy signaling pathway,suggesting its potential for treatment of bone destruction in rheumatoid arthritis.

AIM To study the chemical constituents from the buds of Aralia chinensis L.var.nuda Nakai and their in vitro anti-inflammatory activities.METHODS The 70％ethanol extract from the buds of A.chinensis var.nuda was isolated and purified by silica gel,Sephadex LH-20,ODS and semi-preparative HPLC,then the structures of compounds were identified by physicochemical properties and spectral data.Their anti-inflammatory activities in vitro were evaluated by RAW264.7 model.RESULTS Sixteen compounds were isolated and identified as 4-(2,2-dibutoxyethyl)phenol(1),trans-linalool-3,7-oxide-6-O-β-D-glucopyranoside(2),2'-O-(9Z,12Z,15Z-octadecatrienoyl)glyceryl β-D-galactopyranoside(3),quercetin-3-O-β-D-glucopyranoside(3'→ O-3''')quercetin-3-O-β-D-galactopyranoside(4),syringaresinol-4'-O-β-D-glucopyranoside(5),p-hydroxybenzaldehyde(6),7α-hydroxystigmasterol 3-O-β-D-glucopyranoside(7),trans-p-hydroxy cinnamic acid methyl ester(8),funingensin A(9),3,4-dihydroxy-acetophenone(10),N-acetyltyramine(11),3,4-di-O-caffeoyl quinic acid(12),chlorogenic acid(13),aralia cerebroside(14),caffeic acid methyl ester(15),tetradecanoic acid(16).The IC50values of compounds 8,10,12 and 13 were(22.19±1.59),(35.25±1.30),(13.38±0.72),(15.73±1.16)μmol/L,respectively.CONCLUSION Compound 1 is a new compound,2-13 are isolated from genus Aralia for the first time.Compounds 8,10,12,13 exhibit significant in vitro anti-inflammatory activities.

OBJECTIVE To investigate the inhibitory effects of Bruceine A(BA)on colon cancer and its underlying mechanisms.METHODS Human colon cancer HT-29 and HCT116 cells were treated with various concentrations of BA(0,1,2,5,10,20,40,80 μmol·L-1).Cell viability was assessed using the Cell Counting Kit-8(CCK-8).Flow cytometry,wound healing assays,and Transwell assays were employed to evaluate the effects of BA on cell apoptosis,cell cycle,invasion,and migration.Mo-lecular docking simulations were used to assess the binding of BA to GSK-3β protein,and Western blot analysis was used to examine protein expression related to the cell cycle,epithelial-mesenchymal transition,and the Wnt/β-catenin signaling pathway.An HT-29 cell subcutaneous xenograft mouse model was established.After tumor formation,mice were randomly divided into three groups(six mice per group):a blank group,a low-dose BA group(0.1 mg·kg-1),and a high-dose BA group(0.2 mg·kg-1).Mice were ad-ministered the drug for 19 d,then sacrificed,and tumor tissues were collected.Tumor volume changes over time were observed;Ki67 immunohistochemistry was used to assess cell proliferation in tumor tissues;Western blot analysis of Wnt/β-catenin signaling pathway protein expression was conducted.RESULTS Compared with the blank group,BA could significantly inhibit the proliferation of HT-29 and HCT116 cells,with IC50 values of 10.80 μmol·L-1 and 17.96 μmol·L-1,respectively.Flow cytometry results showed that BA significantly induced apoptosis of HT-29 cells(P<0.01,P<0.001),and arrested the cell cycle at the S phase,accompanied by de-creased expression of cycle-related proteins CDK2 and Cyclin A(P<0.05,P<0.01,P<0.001).BA inhibited cell migration and in-vasion ability(P<0.05,P<0.01,P<0.001),reduced the expression of EMT-related proteins Snail,Vimentin,and N-Cadherin(P<0.01,P<0.001),and upregulated the expression of E-Cadherin protein.In addition,BA inhibited the expression of β-catenin and p-GSK3β proteins.Wnt agonist LiCl could significantly antagonize the anti-colon cancer effect of BA;Wnt inhibitor XAV939 could enhance the anti-colon cancer effect of BA.In the in vivo experiment,compared with the blank group,the tumor volume of the low-dose and high-dose BA groups was significantly reduced(P<0.05,P<0.001).Immunohistochemistry results showed that compared with the blank group,the expression of Ki67 in tumor tissues of the low-dose and high-dose BA groups was significantly reduced(P<0.001).Western blot results further proved that BA inhibited the Wnt/β-catenin signaling pathway.CONCLUSION BA inhibits the viability,invasion,and migration of colon cancer HT-29 cells,induces apoptosis,and causes cell cycle arrest.Additionally,it significantly suppresses the growth of subcutaneous HT-29 cell xenografts in vivo,possibly related to the Wnt/β-catenin signaling pathway.

Objective To propose a Weka-based method for classifying acute lymphoblastic leukemia(ALL)images,aiming to improve ALL cell classification accuracy and stability.Methods Firstly,totally 180 images were randomly selected from ALL-IDB2 subset of Acute Lymphoblastic Leukemia Image Database(ALL-IDB),including 90 images of patients and 90 images of healthy people;secondly,the image preprocessing was carried out using ImageJ software and image features were extracted such as texture,edge and shape;thirdly,image classification was implemented with four classifiers of Weka,including random forest(RF),Bayesian network(BN),J48 decision tree and sequential minimal optimization(SMO),and the key parameters of each classifier were optimized;finally,the performance of the classifiers was verified using 80 independent test images.Results Before parameter optimization,the accuracy of RF,J48 decision tree,BN and SMO classifiers was 94.3％,86.2％,83.6％and 83.0％,respectively.After optimization,the accuracy increased to 95.2％,86.3％,86.3％and 89.7％,respectively.After optimization,RF behaved the best on the independent test set with a classification accuracy of 90.0％,followed by SMO(81.3％),BN(81.3％)and J48 decision tree(75.0％).Conclusion The Weka-based ALL image classification method with a high accuracy is efficient and reliable for automated classification of ALL cell.[Chinese Medical Equipment Journal,2025,46(2):10-15]

Objective:To compare the impact of manual right arm blood pressure measurement with computer-controlled blood pressure meter (CCBPM) on the detection rate of hypertension among elderly individuals.Method:This was a cross-sectional study. Elderly residents undergoing routine health check-up in a village in Jiangxi Province from April to June 2024 were enrolled. Manual blood pressure measurements were performed on the right arm using an electronic sphygmomanometer, while standardized dual-arm synchronized blood pressure assessments were conducted using the CCBPM. Blood pressure data were recorded from three sources: manual measurement on the right arm; the first CCBPM measurement on the arm with higher systolic blood pressure (CCBPM single); and the mean blood pressure of the arm with higher systolic blood pressure from two (if the difference between the first two measurements was less than 5 mmHg, 1 mmHg=0.133 kPa) or three (if the difference exceeded 5 mmHg) CCBPM measurements (CCBPM mean). Hypertension detection rates from these three datasets were compared. Subgroup analysis was performed according to age (60-74 years old as the elderly group,≥75 years old as the senior group) and gender. Intra-group correlation coefficient ( ICC) was analyzed to evaluate the consistency of blood pressure data of different blood pressure measurement methods, and Kappa value was analyzed to evaluate the consistency of hypertension classification determined by different blood pressure measurement methods. Results:A total of 1 498 participants were enrolled, aged (71.30±6.83) years old, including 678 males (45.26%). The blood pressure and heart rate measured by the artificial right arm blood pressure measurement were higher than those measured by CCBPM single (blood pressure: (140.09±17.19)/(82.40±10.93) mmHg vs. (135.31±18.98)/(81.23±10.51) mmHg; heart rate: (75.76±11.72) beats/min vs.(72.94±11.21) beats/min) and CCBPM mean (blood pressure: (140.09±17.19)/(82.40±10.93) mmHg vs. (134.64±18.39)/(80.28±9.78) mmHg; heart rate: (75.76±11.72) beats/min vs. (72.87±10.70) beats/min, all P<0.05). The detection rate of hypertension determined by CCBPM mean was significantly lower than that of artificial right arm (40.25% (603/1 498) vs.54.34% (814/1 498)) and CCBPM single (40.25% (603/1 498) vs. 44.79% (671/1 498), all P<0.05). Subgroup analysis showed that that the detection rate of hypertension determined by CCBPM mean was lower than that measured by artificial right arm regardless of gender and age (all P<0.05). The consistency of blood pressure data between artificial right arm and the CCBPM mean was moderate (systolic blood pressure: ICC=0.70; diastolic blood pressure: ICC=0.62), with less consistent classification ( Kappa=0.37). The consistency of blood pressure data between CCBPM single and CCBPM mean is extremely high (systolic blood pressure: ICC=0.94; diastolic blood pressure: ICC=0.91), with highly consistent classification ( Kappa=0.74). Conclusions:Artificial right arm blood pressure measurement in physical examinations may overestimate the hypertension detection rate, and the standardized dual-arm synchronized blood pressure measurement using CCBPM can reduce irregular blood pressure measurement.

Objective To explore the effect and mechanism of lentinan(LNT)on hepatic tissue ferroptosis in mice exposed to sodium arsenite(SA).Methods C57BL/6N male mice were exposed to SA low-dose,SA high-dose,and LNT intervention combined with SA high-dose,then,hematoxylin and eosin(HE)staining was applied to assess pathological liver tissue damage;Enzyme-linked immunosorbent and Western blot were used to detect the content or expression of tumor necrosis factor α(TNFα),interleukin-6(IL-6),ferritinophagy or ferroptosis biomarkers.Results Compared with the control group,SA exposure induced the elevated levels of TNFα,IL-6,ferritinophagy biomarker ferritin heavy chain 1(FTH1)and microtubule-associated protein 1 light chain 3B(MAP1LC3B)in mice liver tissue,while levels the ferroptosis biomarker GPX4 decreased(P＜0.05).Compared with SA high-dose groups,LNT intervention showed the reduced pathological liver damage and the downregulated levels of TNFα,IL-6,FTH1,and MAP1LC3B,while the level of GPX4 upregulated(P＜0.05).Western blot experiment showed that LNT intervention antagonized the upregulated levels of FTH1,and autophagy biomarker LC3B/A,and antagonized the increased co-expressions of FTH1 with LC3B or Ub protein in SA high-dose group(P＜0.05).Conclusions LNT antagonizes SA-exposed hepatic pathological injury and ferroptosis in mice,possibly associated with inhibition of TNFα-ferritinophagy signaling.

Poly(lactic-co-glycolide)(PLGA)is a key excipient in long-acting sustained-release preparations,and its degradation properties directly affect the drug release behavior.In this study,PLGA microspheres were prepared by microfluidic techniques,and the morphology changes of the microspheres were observed by scanning electron microscopy(SEM).In alkaline environment,due to the accelerated hydrolysis of ester bonds,the surface of the microspheres was rapidly dissolved and eroded,and the degradation rate was significantly higher than that in acidic environment.High temperature accelerated the degradation of PLGA microspheres.Under neutral and alkaline conditions,the microspheres showed aggregation and adhesion.Under acidic conditions,the microspheres gradually decomposed into irregular fragments.The high ionic strength further promoted the surface corrosion of the microspheres,especially under extreme pH conditions.Simultaneously,PLGA microspheres encapsulating coumarin were prepared to simulate the microsphere formulation.The release rate of coumarin after degradation of the microspheres under different conditions was observed by measuring the absorbance with ultraviolet-visible spectrophotometry.The results were consistent with those of the blank microspheres.This study revealed that the degradation of PLGA microspheres was significantly pH-dependent,temperature sensitive and ion strength responsive.These findings not only helped to understand and optimize the long-term stability and controlled release performance of drug-carrying microspheres,but also provided a theoretical basis for further improvement of PLGA-based drug carrier design.