Objective To evaluate the effect of two different acid etching agents on the bonding strength between deciduous enamel and composite resin at different etching times.Methods Seventy primary incisors were made into test specimens and randomly divided into 7 groups(n=10),6 experimental groups,and 1 blank control group.The specimens were divided into groups and etchedwith two different acid etching agents(35％H3PO4,15％HCl)for different durations(15,30,60 s).After bonding with the composite resin,shear force testing was performed.Additionally,two deciduous molar dental crowns were cut vertically along the gingival axis,with each tooth divided into seven sections and randomly assigned to seven groups.Enamel acid etched specimens were produced and the surface morphology was observed using scanning electron microscopy.Finally,24 deciduous molars were randomly divided into 3 groups(n=8)according to the etching time(15,30,60 s),and each tooth was vertically cut into 2 pieces along the buccal lingual direction,namely the phosphoric acid group and the hydrochloric acid group,and etched for the same time(n=8),for a total of 6 groups.They were bonded with resin and made into specimens.The specimen was cut vertically to the bonding surface into 2 parts.One measurement point was selected for each part,and each group had a total of 16 measurement values.Resin protrusion length was measured under scanning electron microscopy and was statistically analyzed.Results When comparing acid etching for 15,30,and 60 s,the bonding strength of the phosphate group was higher than that of the hydrochloric acid group.Under scanning electron microscopy,there was no significant difference in the acid etching mode between the two groups,both of which were type 2 acid etching modes dissolved along the glaze column direction.At 15 seconds of acid etching,the enamel surface was uneven,and continuous and uniform dissolution ap-peared at 30 seconds.The length of resin protrusion increased with the increase of acid etching time,and the phosphoric acid group was greater than the hydrochloric acid group at 15,30,and 60 s compared between groups.Conclusion The etching time is positively correlated with the shear bonding strength.The best bonding effect is achieved when deciduous teeth are etched with 35％ H3PO4 for 60 seconds,and the effect of 35％phosphoric acid etching on deciduous tooth enamel is better than that of 15％hydrochloric acid.

Objective To investigate the correlation between peripheral blood CD14+CD16+monocytes and IgG N-glycosyl levels and disease activity in patients with systemic lupus erythematosus(SLE).Methods A total of 109 SLE patients admitted to Xingtai Cental Hospital from August 2021 to November 2024 were retrospectively selected as the study objects.According to SLE disease activity index(SLE-DAI),the patients were divided into active group(n=52)and stable group(n=57).In addition,56 patients who underwent physical examination during the same period were selected as the control group.Clinical data of patients were collected,CD14+CD16+mononuclear cells were detected by flow cytometry(FCM),and IgG N-glycosyl levels were detected by hydrophilic interaction chromatography-mass spectrometry.Logistic regression analysis was performed to analyze the influencing factors of SLE-DAI,and multicollinearity test[variance inflation factor(VIF)]was performed for independent variables.The prediction model of disease activity was constructed.The effectiveness of the predictive model was evaluated by describing the recviver operator characteristic(ROC)curve and calculating the area under the curve(AUC)value.Hosmer-Lemeshow goodness-of-fit test predicted the calibration degree of the model.Results The levels of WBC,Hb,PLT,ALB,complement C3 and complement C4 in control group were higher than those in SLE group(t=8.917～22.171),and the levels of CRP were lower than those in SLE group(t=-17.359),with differences were statistical significance(all P<0.05).The CRP level and the proportion of CD14+CD16+mononuclear cells in the active group were higher than those in the stable group,and the differences were statistically significant(t=5.449,11.112,all P<0.05).The IgG glycosylation characteristics of galactosylation,sialylation and N-acetylglucosamine modification were lower than those in the stable group,and the differences were statistical significance(Z=-2.432～-0.158,all P<0.05).Spearman correlation analysis showed that the proportion of CD14+CD16+monocytes was significantly negatively correlated with IgG galactosylation,sialylation level and bisection N-acetylglucosamine modification(r=-0.656,-0.531,-0.608,all P<0.01).CD14+CD16+monocyte ratio was positively correlated with SLE-DIA(r=0.581,all P<0.01).IgG galactosylation,sialylation levels and bisection N-acetylglucosamine modification were negatively correlated with SLE-DIA(r=-0.645,-0.609,-0.503,all P<0.01).Logistic regression analysis showed that CRP>8.21mg/L,CD14+CD16+≥16.17%,sialylation<22.05%and isotropic N-acetylglucosamine modification<16.53%were independent risk factors for disease activity in SLE patients(Wald χ2=4.471～12.811,all P<0.05).The VIF values of the above independent variables were all less than 10.By establishing the Logistic regression prediction model and drawing the ROC curve,the AUC value for diagnosing SLE disease activity was 0.821(95%CI:0.733～0.905),the sensitivity,specificity and the Yodon index were 85.37%,75.67%,0.677,respectively.and the P values of Hosmer-Lemeshow goodness-of-fit test models were 0.568,respectively.Conclusion The proportion of CD14+CD16+monocytes in peripheral blood of SLE patients increase significantly,and the level of IgG glycosylation characteristics decrease,both of which are correlated with SLE-DIA.The predictive model constructed based on the two had a good ability to distinguish SLE-DIA from inactive state,with high sensitivity and moderate specificity conducive to early clinical recognition,and the model fitting effect is good.SLE-DIA can be evaluated more accurately.

Objective To evaluate the effect of two different acid etching agents on the bonding strength between deciduous enamel and composite resin at different etching times.Methods Seventy primary incisors were made into test specimens and randomly divided into 7 groups(n=10),6 experimental groups,and 1 blank control group.The specimens were divided into groups and etchedwith two different acid etching agents(35％H3PO4,15％HCl)for different durations(15,30,60 s).After bonding with the composite resin,shear force testing was performed.Additionally,two deciduous molar dental crowns were cut vertically along the gingival axis,with each tooth divided into seven sections and randomly assigned to seven groups.Enamel acid etched specimens were produced and the surface morphology was observed using scanning electron microscopy.Finally,24 deciduous molars were randomly divided into 3 groups(n=8)according to the etching time(15,30,60 s),and each tooth was vertically cut into 2 pieces along the buccal lingual direction,namely the phosphoric acid group and the hydrochloric acid group,and etched for the same time(n=8),for a total of 6 groups.They were bonded with resin and made into specimens.The specimen was cut vertically to the bonding surface into 2 parts.One measurement point was selected for each part,and each group had a total of 16 measurement values.Resin protrusion length was measured under scanning electron microscopy and was statistically analyzed.Results When comparing acid etching for 15,30,and 60 s,the bonding strength of the phosphate group was higher than that of the hydrochloric acid group.Under scanning electron microscopy,there was no significant difference in the acid etching mode between the two groups,both of which were type 2 acid etching modes dissolved along the glaze column direction.At 15 seconds of acid etching,the enamel surface was uneven,and continuous and uniform dissolution ap-peared at 30 seconds.The length of resin protrusion increased with the increase of acid etching time,and the phosphoric acid group was greater than the hydrochloric acid group at 15,30,and 60 s compared between groups.Conclusion The etching time is positively correlated with the shear bonding strength.The best bonding effect is achieved when deciduous teeth are etched with 35％ H3PO4 for 60 seconds,and the effect of 35％phosphoric acid etching on deciduous tooth enamel is better than that of 15％hydrochloric acid.

Objective To investigate the correlation between peripheral blood CD14+CD16+monocytes and IgG N-glycosyl levels and disease activity in patients with systemic lupus erythematosus(SLE).Methods A total of 109 SLE patients admitted to Xingtai Cental Hospital from August 2021 to November 2024 were retrospectively selected as the study objects.According to SLE disease activity index(SLE-DAI),the patients were divided into active group(n=52)and stable group(n=57).In addition,56 patients who underwent physical examination during the same period were selected as the control group.Clinical data of patients were collected,CD14+CD16+mononuclear cells were detected by flow cytometry(FCM),and IgG N-glycosyl levels were detected by hydrophilic interaction chromatography-mass spectrometry.Logistic regression analysis was performed to analyze the influencing factors of SLE-DAI,and multicollinearity test[variance inflation factor(VIF)]was performed for independent variables.The prediction model of disease activity was constructed.The effectiveness of the predictive model was evaluated by describing the recviver operator characteristic(ROC)curve and calculating the area under the curve(AUC)value.Hosmer-Lemeshow goodness-of-fit test predicted the calibration degree of the model.Results The levels of WBC,Hb,PLT,ALB,complement C3 and complement C4 in control group were higher than those in SLE group(t=8.917～22.171),and the levels of CRP were lower than those in SLE group(t=-17.359),with differences were statistical significance(all P<0.05).The CRP level and the proportion of CD14+CD16+mononuclear cells in the active group were higher than those in the stable group,and the differences were statistically significant(t=5.449,11.112,all P<0.05).The IgG glycosylation characteristics of galactosylation,sialylation and N-acetylglucosamine modification were lower than those in the stable group,and the differences were statistical significance(Z=-2.432～-0.158,all P<0.05).Spearman correlation analysis showed that the proportion of CD14+CD16+monocytes was significantly negatively correlated with IgG galactosylation,sialylation level and bisection N-acetylglucosamine modification(r=-0.656,-0.531,-0.608,all P<0.01).CD14+CD16+monocyte ratio was positively correlated with SLE-DIA(r=0.581,all P<0.01).IgG galactosylation,sialylation levels and bisection N-acetylglucosamine modification were negatively correlated with SLE-DIA(r=-0.645,-0.609,-0.503,all P<0.01).Logistic regression analysis showed that CRP>8.21mg/L,CD14+CD16+≥16.17%,sialylation<22.05%and isotropic N-acetylglucosamine modification<16.53%were independent risk factors for disease activity in SLE patients(Wald χ2=4.471～12.811,all P<0.05).The VIF values of the above independent variables were all less than 10.By establishing the Logistic regression prediction model and drawing the ROC curve,the AUC value for diagnosing SLE disease activity was 0.821(95%CI:0.733～0.905),the sensitivity,specificity and the Yodon index were 85.37%,75.67%,0.677,respectively.and the P values of Hosmer-Lemeshow goodness-of-fit test models were 0.568,respectively.Conclusion The proportion of CD14+CD16+monocytes in peripheral blood of SLE patients increase significantly,and the level of IgG glycosylation characteristics decrease,both of which are correlated with SLE-DIA.The predictive model constructed based on the two had a good ability to distinguish SLE-DIA from inactive state,with high sensitivity and moderate specificity conducive to early clinical recognition,and the model fitting effect is good.SLE-DIA can be evaluated more accurately.

In order to obtain polyclonal antibodies against the fibrillar(Fiber)protein of fowl ade-novirus serotype 11(FAdV-11)and investigate its cross-reactivity to different serotypes of FAdV Fiber,the gene encoding the FAdV-11 Fiber protein was cloned into a prokaryotic expression vec-tor pET-32a by homologous recombination technology,then the plasmid was transformed into BL21(DE3)receptor cells,and the purified recombinant protein was used as an immunogen to im-munize rabbits to prepare polyclonal antibody after induced expression,and the cross-reactivity of the polyclonal antibody against different serotypes of FAdV Fiber proteins was identified by West-ern blot and indirect immunofluorescence(IFA).The results showed that the His-FAdV-11-Fiber recombinant protein was mainly expressed as inclusion bodies and was well expressed.Western blot and IFA showed that the prepared polyclonal antibody reacted with the Fiber proteins of FAdV-8a,FAdV-8b,and FAdV-11,but did not with the 2 Fiber of FAdV-4(Fiber 1 and Fiber 2)proteins.In conclusion,in this study,we successfully prepared rabbit polyclonal antibodies against FAdV-11 Fiber and showed that it specifically recognized the Fiber proteins of FAdV-8a,FAdV-8b and FAdV-11,which lays the foundation for further establishment of serological differential diag-nosis of FAdV-11.

Breast cancer is the most common cancer in women and one of the deadliest cancers worldwide.According to the distribution of tumor tissue,breast cancer can be divided into invasive and non-invasive forms.The cancer cells in invasive breast cancer pass through the breast and through the immune system or systemic circulation to different parts of the body,forming metastatic breast cancer.Drug resistance and distant metastasis are the main causes of death from breast cancer.Research on breast cancer has attracted extensive attention from researchers.In vitro construction of tumor models by tissue engineering methods is a common tool for studying cancer mechanisms and anticancer drug screening.The tumor microenvironment consists of cancer cells and various types of stromal cells,including fibroblasts,endothelial cells,mesenchymal cells,and immune cells embedded in the extracellular matrix.The extracellular matrix contains fibrin proteins(such as types Ⅰ,Ⅱ,Ⅲ,Ⅳ,Ⅵ,and Ⅹcollagen and elastin)and glycoproteins(such as proteoglycan,laminin,and fibronectin),which are involved in cell signaling and binding of growth factors.The current traditional two-dimensional(2D)tumor models are limited by the growth environment and often cannot accurately reproduce the heterogeneity and complexity of tumor tissues in vivo.Therefore,in recent years,research on three-dimensional(3D)tumor models has gradually increased,especially 3D bioprinting models with high precision and repeatability.Compared with a 2D model,the 3D environment can better simulate the complex extracellular matrix components and structures in the tumor microenvironment.Three-dimensional models are often used as a bridge between 2D cellular level experiments and animal experiments.Acellular matrix,gelatin,sodium alginate,and other natural materials are widely used in the construction of tumor models because of their excellent biocompatibility and non-immune rejection.Here,we review various natural scaffold materials and construction methods involved in 3D tissue-engineered tumor models,as a reference for research in the field of breast cancer.

OBJECTIVE: To explore the effect and mechanism of miR-125a-5p targeted regulation of scavenger receptor B1 (Scarb1) gene on anoxia/reoxygenation injury of rat cardiomyocytes. METHODS: H9c2 rat cardiomyocytes were randomly divided into blank control group, hypoxia/reoxygenation group, transfection control group and mir-125a-5p transfection group. The expression of miR-125a-5p, cardiomyocyte viability, apoptosis rate, ATP content and the expression of Scarb1, Cyt C, Bax, Bcl-2 and NF-κB signaling pathway related proteins were determined. Target gene of miR-125a-5p was predicted with Targetscan software, and the targeting of miR-125a-5p on Scarb1 was verified by double luciferase reporter gene experiment. RESULTS: Compared with the blank control group, the expression of miR-125a-5p, Bax, Cyt C and the apoptotic rate of cardiomyocytes in the hypoxia/reoxygenation group were significantly increased (P<0.05), while the expression of Scarb1, Bcl-2 and the content of ATP were significantly decreased (P<0.05). Compared with the control group, the situation of mir-125a-5p transfection group was just the opposite. Double luciferase reporter gene experiment has confirmed Scarb1 to be the target of miR-125a-5p. Hypoxia/reoxygenation can promote the expression of NF-κB p65, C-myc and Cyclin D1 in cardiomyocytes, while down-regulating the expression of miR-125a-5p can inhibit the expression of such proteins. CONCLUSION Hypoxia/reoxygenation can induce the expression of miR-125a-5p in rat cardiomyocytes. Inhibition of miR-125a-5p can protect cardiomyocytes from hypoxia/reoxygenation by up-regulating the expression of Scarb1. The mechanism may be related to the inhibition of activation of NF-κB signaling pathway.

Objective: To investigate the dynamic changes of antibodies induced by leptospiral vaccines. Methods: Antigens for antibody detection were screened out. ELISA was used to analyze antibody responses induced at different time points after immunizing guinea pigs with different batches of leptospiral vaccines from different manufacturers. To investigate the relationship between antibody responses induced by leptospiral vaccines and their protective effects in animal model, guinea pigs were challenged with Leptospira after immunization. Results: There was no significant antigen-antibody reaction between the LigA protein or Patoc Ⅰ antigen and the serum samples of guinea pigs immunized with leptospiral vaccines. Notable IgG and IgM antibody reactions were observed in all vaccination groups when using bacterial proteins from seven Leptospira reference strains which were used for the preparation of leptospiral vaccines as envelope antigens. Antigen-specific IgG antibodies peaked at 35 d after the last immunization, and the highest peak of antigen-specific IgM antibodies was reached 11 d after the last immunization. Results of the challenge test showed that non-diluted leptospiral vaccines induced significant IgG and IgM antibody reactions in guinea pigs as compared with those diluted three or nine times, showing good protective effects. Conclusions Analysis of the dynamic changes of antibodies induced by leptospiral vaccines revealed that there was correlation between the induced serum antibody responses and the protective effects. This study provided reference for further study on alternative methods for evaluating leptospiral vaccine potency.

Collaboration of c-KIT mutations with AML1-ETO (AE) has been demonstrated to induce t(8; 21) acute myeloid leukemia (AML). Targeted therapies designed to eliminate AE and c-KIT oncoproteins may facilitate effective treatment of t(8; 21) AML. Homoharringtonine (HHT) features activity against tumor cells harboring c-KIT mutations, whereas oridonin can induce t(8; 21) AML cell apoptosis and AE cleavage. Therefore, studies should explore the efficacy of combination therapy with oridonin and HHT in t(8; 21) AML. In this study, we investigated the synergistic effects and mechanism of oridonin combined with HHT in t(8; 21) AML cell line and mouse model. The two drugs synergistically inhibited cell viability and induced significant mitochondrial membrane potential loss and apoptosis. Oridonin and HHT induced significant downregulation of c-KIT and its downstream signaling pathways and promoted AE cleavage. HHT increased intracellular oridonin concentration by modulating the expressions of MRP1 and MDR1, thus enhancing the effects of oridonin. The combination of oridonin and HHT prolonged t(8; 21) leukemia mouse survival. In conclusion, oridonin and HHTexert synergistic effects against t(8; 21) leukemia in vivo and in vitro, thereby indicating that their combination may be an effective therapy for t(8; 21) leukemia.

Objective:To investigate the chemical constituents in the rattan of Rubia argyi L.. Methods:The air-dried rattan of Rubia argyi L. was powdered and extracted three times by 75% ethanol with refluxing. After removing the solvent under the reduced pressure,the crude extract was dissolved in water,and then filtrated and extracted by petroleum ether and ethyl acetate to obtain crude extract after removing petroleum ether and ethyl acetate. The compounds were isolated and purified by silica gel column chromatogra-phy,reversed-phase silica gel column chromatography and Sephadex LH-20 gel column chromatography,and then identified based on physicochemical properties and spectral analysis(1 H-NMR and 13C-NMR). Results:Totally 13 compounds were isolated from the rat-tan of Rubia argyi L.,and characterized as secoisolariciresinol(1),xanthopurpurin(2),daucosterol(3),dehydroabietic acid(4), 2-hydroxy-1-methoxy-anthraquinone(5),β-sitosterol(6),lirioresinol A(7),2-hydroxy-7-methyl-9,10-anthraquinone(8),strych-novoline (9), ciwujiatone (10), 3,4-divanillyltetrahydrofuran (11), 2-(4-hydroxypheny) -6-(3-methoxy-4-hydroxyphenyl)-3,7-dioxabicyclo[3,3,0]octane (12), and (6S,9R)-vomifoliol (13).Conclusion: The compounds 1-13 are isolated from the rattan of Rubia argyi L. for the first time and the compounds 1,2,4,5 and 7-13 are first isolated from Rubia L..