Objective:To investigate effects of picroside Ⅱ(PⅡ)on proliferation,apoptosis and immune escape of lung cancer cells by regulating C-C motif chemokine ligand 2(CCL2)/C-C motif chemokine receptor 2(CCR2)signaling axis.Methods:Human lung cancer cells NCI-H292 were cultured and treated with 0,5,10,20,40 and 80 μmol/L PⅡ,MTT method was applied to detect cell viability.Experiment was separated into control group,low,medium and high concentrations PⅡ groups(PⅡ-L,PⅡ-M,PⅡ-H,10,20 and 40 μmol/L PⅡ),high concentration PⅡ+CCL2 overexpression negative control group(PⅡ-H+pcDNA-NC,40 μmol/L PⅡ+pcDNA-NC)and high concentration PⅡ+CCL2 overexpression group(PⅡ-H+CCL2,40 μmol/L PⅡ+pcDNA-CCL2).EdU method was applied to measure cell proliferation;flow cytometry was applied to measure cell apoptosis;immunoblotting was applied to determine expressions of CCL2,CCR2,B-cell lymphoma-2(Bcl-2)and Bcl-2 associated X protein(Bax).Lung can-cer cells in each group were co-cultured with CD8+T cells,Trypan blue staining was applied to measure CD8+T cell viability;ELISA was applied to determine levels of programmed death receptor-ligand 1(PD-L1),IL-10,IFN-γ and TGF-β.Results:Compared with 0 μmol/L,cell viability treated with 10,20,40 and 80 μmol/L PⅡ were significantly reduced(P<0.05),and 10,20 and 40 μmol/L PⅡ were selected for subsequent experiments.Compared with control group,positive rate of EdU and expressions of Bcl-2,CCL2 and CCR2 in PⅡ-L group,PⅡ-M group and PⅡ-H group were decreased sequentially(P<0.05),while apoptosis rate and expression of Bax were increased sequentially(P<0.05).Compared with PⅡ-H+pcDNA-NC group,positive rate of EdU and expressions of Bcl-2,CCL2 and CCR2 in PⅡ-H+CCL2 group were increased obviously(P<0.05),while apoptosis rate and expression of Bax were de-creased significantly(P<0.05).After co-culturing with CD8+T cells,compared with control group,levels of IL-10,TGF-β and PD-L1 in PⅡ-L group,PⅡ-M group and PⅡ-H group were decreased sequentially(P<0.05),while CD8+T cell viability and level of IFN-γ were increased sequentially(P<0.05).Compared with PⅡ-H+pcDNA-NC group,levels of IL-10,TGF-β and PD-L1 in PⅡ-H+CCL2 group were increased obviously(P<0.05),while CD8+T cell viability and level of IFN-γ were reduced significantly(P<0.05).Conclu-sion:PⅡ may inhibit proliferation and immune escape of lung cancer cells,and promote cell apoptosis by inhibiting CCL2-CCR2 sig-naling axis.

Objective To investigate the effects of asiaticoside(AS)on epithelial-mesenchymal transition(EMT)and radiotherapy sensitivity of esophageal cancer(EC)cells by its mechanism of regulating the hypoxia inducible factor-1α(HIF-1α)/vascular endothelial growth factor(VEGF)signaling pathway.Methods EC9706 cells were subjected to different concentrations of AS or different doses of radiation.Methyl thiazolyl tetrazolium method was used to detect cell proliferation and calculate the half-maximal inhibitory con-centration.EC9706 cells were divided into a control group,radiology group(X-ray irradiation),AS group,combined group(AS+X-ray irradiation),and activator group(AS+X-ray irradiation+HIF-1α/VEGF pathway activator dimethyloxallyl glycine).Plate cloning experi-ments were conducted to detect sensitivity,and Transwell assays were used to detect cell migration and invasion.Flow cytometry helped detect apoptosis,and real-time quantitative polymerase chain reaction detected the expression of HIF-1α and VEGF mRNA.Western blotting method was used to detect the expression of matrix metalloproteinase-2(MMP-2),vimentin,E-cadherin,Bcl-2 associated X protein(Bax),HIF-1α,and VEGF proteins.Results With the increase of AS concentration and radiation dose,the cell viability of EC9706 cells gradually decreased;compared with the control group,the survival fraction;the numbers of cells that had migrated and invaded;the expression of HIF-1α and VEGF mRNA;and the expression of MMP-2,vimentin,HIF-1α,and VEGF in the radiology group and AS group were reduced;further,the apoptosis rate and the expression of E-cadherin and Bax were increased(P＜0.05).Compared with the radiology group and AS group,the survival fraction;the numbers of cells that had migrated and invaded;the expression of HIF-1αand VEGF mRNA;and the expressions of MMP-2,vimentin,HIF-1α,and VEGF in the combined group were reduced;the apoptosis rate and the expression of E-cadherin and Bax were increased(P＜0.05).In comparison with the combined group,the changes in the above indicators in the activator group were reversed(P＜0.05).Conclusion AS may inhibit EMT by inhibiting the HIF-1α/VEGF signaling pathway,thus enhancing the radiotherapy sensitivity of EC cells.

Objective:To investigate effects of picroside Ⅱ(PⅡ)on proliferation,apoptosis and immune escape of lung cancer cells by regulating C-C motif chemokine ligand 2(CCL2)/C-C motif chemokine receptor 2(CCR2)signaling axis.Methods:Human lung cancer cells NCI-H292 were cultured and treated with 0,5,10,20,40 and 80 μmol/L PⅡ,MTT method was applied to detect cell viability.Experiment was separated into control group,low,medium and high concentrations PⅡ groups(PⅡ-L,PⅡ-M,PⅡ-H,10,20 and 40 μmol/L PⅡ),high concentration PⅡ+CCL2 overexpression negative control group(PⅡ-H+pcDNA-NC,40 μmol/L PⅡ+pcDNA-NC)and high concentration PⅡ+CCL2 overexpression group(PⅡ-H+CCL2,40 μmol/L PⅡ+pcDNA-CCL2).EdU method was applied to measure cell proliferation;flow cytometry was applied to measure cell apoptosis;immunoblotting was applied to determine expressions of CCL2,CCR2,B-cell lymphoma-2(Bcl-2)and Bcl-2 associated X protein(Bax).Lung can-cer cells in each group were co-cultured with CD8+T cells,Trypan blue staining was applied to measure CD8+T cell viability;ELISA was applied to determine levels of programmed death receptor-ligand 1(PD-L1),IL-10,IFN-γ and TGF-β.Results:Compared with 0 μmol/L,cell viability treated with 10,20,40 and 80 μmol/L PⅡ were significantly reduced(P<0.05),and 10,20 and 40 μmol/L PⅡ were selected for subsequent experiments.Compared with control group,positive rate of EdU and expressions of Bcl-2,CCL2 and CCR2 in PⅡ-L group,PⅡ-M group and PⅡ-H group were decreased sequentially(P<0.05),while apoptosis rate and expression of Bax were increased sequentially(P<0.05).Compared with PⅡ-H+pcDNA-NC group,positive rate of EdU and expressions of Bcl-2,CCL2 and CCR2 in PⅡ-H+CCL2 group were increased obviously(P<0.05),while apoptosis rate and expression of Bax were de-creased significantly(P<0.05).After co-culturing with CD8+T cells,compared with control group,levels of IL-10,TGF-β and PD-L1 in PⅡ-L group,PⅡ-M group and PⅡ-H group were decreased sequentially(P<0.05),while CD8+T cell viability and level of IFN-γ were increased sequentially(P<0.05).Compared with PⅡ-H+pcDNA-NC group,levels of IL-10,TGF-β and PD-L1 in PⅡ-H+CCL2 group were increased obviously(P<0.05),while CD8+T cell viability and level of IFN-γ were reduced significantly(P<0.05).Conclu-sion:PⅡ may inhibit proliferation and immune escape of lung cancer cells,and promote cell apoptosis by inhibiting CCL2-CCR2 sig-naling axis.

Objective To investigate the effects of asiaticoside(AS)on epithelial-mesenchymal transition(EMT)and radiotherapy sensitivity of esophageal cancer(EC)cells by its mechanism of regulating the hypoxia inducible factor-1α(HIF-1α)/vascular endothelial growth factor(VEGF)signaling pathway.Methods EC9706 cells were subjected to different concentrations of AS or different doses of radiation.Methyl thiazolyl tetrazolium method was used to detect cell proliferation and calculate the half-maximal inhibitory con-centration.EC9706 cells were divided into a control group,radiology group(X-ray irradiation),AS group,combined group(AS+X-ray irradiation),and activator group(AS+X-ray irradiation+HIF-1α/VEGF pathway activator dimethyloxallyl glycine).Plate cloning experi-ments were conducted to detect sensitivity,and Transwell assays were used to detect cell migration and invasion.Flow cytometry helped detect apoptosis,and real-time quantitative polymerase chain reaction detected the expression of HIF-1α and VEGF mRNA.Western blotting method was used to detect the expression of matrix metalloproteinase-2(MMP-2),vimentin,E-cadherin,Bcl-2 associated X protein(Bax),HIF-1α,and VEGF proteins.Results With the increase of AS concentration and radiation dose,the cell viability of EC9706 cells gradually decreased;compared with the control group,the survival fraction;the numbers of cells that had migrated and invaded;the expression of HIF-1α and VEGF mRNA;and the expression of MMP-2,vimentin,HIF-1α,and VEGF in the radiology group and AS group were reduced;further,the apoptosis rate and the expression of E-cadherin and Bax were increased(P＜0.05).Compared with the radiology group and AS group,the survival fraction;the numbers of cells that had migrated and invaded;the expression of HIF-1αand VEGF mRNA;and the expressions of MMP-2,vimentin,HIF-1α,and VEGF in the combined group were reduced;the apoptosis rate and the expression of E-cadherin and Bax were increased(P＜0.05).In comparison with the combined group,the changes in the above indicators in the activator group were reversed(P＜0.05).Conclusion AS may inhibit EMT by inhibiting the HIF-1α/VEGF signaling pathway,thus enhancing the radiotherapy sensitivity of EC cells.

Proteolysis-targeting chimeras (PROTACs) have shown considerable therapeutic potential across diverse fields such as cancer, inflammation, and neurodegenerative diseases, with numerous candidates already progressing into clinical trials. More recently, their application in antiviral therapy has been rapidly gaining momentum. This review systematically outlines the mechanistic foundations and design principles of PROTACs, highlights recent advances targeting coronaviruses (including SARS-CoV-2), hepatitis C virus, human immunodeficiency virus, and influenza viruses, and critically assesses key challenges—particularly the limited diversity of E3 ligase ligands, suboptimal oral bioavailability, and the lack of integrated platforms for druggability evaluation. Looking ahead, innovations in ligand discovery, pathway modulation, delivery technologies, and conditionally activated PROTAC designs are anticipated to overcome these barriers, ushering in a new era of precise and effective antiviral therapeutics.

Objective:To evaluate the left ventricular（LV）function in heart transplant（HTx）patients with post-transplant diabetes（PTDM），and to examine the relevance of PTDM and LV function to the patient's prognosis.Methods:Two hundred and thirteen adult HTx patients who underwent echocardiography at Union Hospital，Tongji Medical College，Huazhong University of Science and Technology between January 2018 and January 2022 were prospectively included. The patients were divided into PTDM group（ n=86）and Non-PTDM group（ n=127）. LV function parameters were acquired using conventional and two-dimensional speckle-tracking echocardiography（2D-STE），and were compared between the two groups. The primary endpoints included all-cause mortality or transplant-related readmission. Results:Compared with Non-PTDM group，the LV mass of PTDM group was higher，the LV ejection fraction，LV global longitudinal strain（GLS），peak systolic global longitudinal strain rate，and early diastolic global longitudinal strain rate（dGLSr）were lower（all P＜0.05）. After a median follow-up period of 37.6（29.3）months，27 patients experienced clinical events. A multivariate analysis revealed that PTDM（ HR=2.198，95% CI=1.018-4.743， P=0.045）and low GLS（ HR=6.456，95% CI=2.889-14.426， P＜0.001）were independent predictors of adverse clinical events after adjustment for dGLSr，body mass index and age. After subdividing the two groups into 4 subgroups by the cutoff value of GLS（16.5%），the prognosis was worst for HTx patients with PTDM and low GLS. Conclusions:HTx patients with PTDM have worse LV systolic and diastolic function than those without PTDM. Management of HTx patients with PTDM may be improved using GLS guidance.

Objective:To evaluate the left ventricular（LV）function in heart transplant（HTx）patients with post-transplant diabetes（PTDM），and to examine the relevance of PTDM and LV function to the patient's prognosis.Methods:Two hundred and thirteen adult HTx patients who underwent echocardiography at Union Hospital，Tongji Medical College，Huazhong University of Science and Technology between January 2018 and January 2022 were prospectively included. The patients were divided into PTDM group（ n=86）and Non-PTDM group（ n=127）. LV function parameters were acquired using conventional and two-dimensional speckle-tracking echocardiography（2D-STE），and were compared between the two groups. The primary endpoints included all-cause mortality or transplant-related readmission. Results:Compared with Non-PTDM group，the LV mass of PTDM group was higher，the LV ejection fraction，LV global longitudinal strain（GLS），peak systolic global longitudinal strain rate，and early diastolic global longitudinal strain rate（dGLSr）were lower（all P＜0.05）. After a median follow-up period of 37.6（29.3）months，27 patients experienced clinical events. A multivariate analysis revealed that PTDM（ HR=2.198，95% CI=1.018-4.743， P=0.045）and low GLS（ HR=6.456，95% CI=2.889-14.426， P＜0.001）were independent predictors of adverse clinical events after adjustment for dGLSr，body mass index and age. After subdividing the two groups into 4 subgroups by the cutoff value of GLS（16.5%），the prognosis was worst for HTx patients with PTDM and low GLS. Conclusions:HTx patients with PTDM have worse LV systolic and diastolic function than those without PTDM. Management of HTx patients with PTDM may be improved using GLS guidance.

OBJECTIVE To investigate the intervention effect and potential mechanism of breviscapine on hepatic fibrosis （HF） in rats based on the transforming growth factor-β（1 TGF-β1）/Smad2/extracellular signal-regulated protein kinase 1（ERK1） and Kelch-like epichlorohydrin-associated protein 1（Keap1）/nuclear factor-erythroid 2-related factor 2（Nrf2）/heme oxygenase-1（HO-1） pathways. METHODS Totally 60 rats were randomly divided into normal control group， model group， breviscapine low-dose， medium-dose and high-dose groups （5.4， 10.8， 21.6 mg/kg）， and colchicine group （positive control， 0.45 mg/kg）， with 10 rats in each group， half male and half female. Except for the normal control group， HF model of the other groups was induced by carbon tetrachloride. Subsequently， each drug group was given corresponding medicine by gavage once a day for 28 days. The liver appearance of rats in each group was observed and their liver coefficients were calculated. The levels of alanineaminotransferase （ALT） and aspartate aminotransferase （AST）in serum， those of ALT， AST， superoxide dismutase （SOD），malondialdehyde （MDA） and glutathione peroxidase （GSH- Px） in liver tissue were detected. The liver tissue inflammatory and fibrotic changes were observed. The protein and mRNA expressions of TGF-β1， Smad2， ERK1， Nrf2， Keap1 and HO-in liver tissue were detected. RESULTS Compared with the normal control group， the model group showed large areas of white nodular lesions in the liver， obvious inflammatory cell infiltration and collagen fiber deposition. The body weight， the levels of SOD and GSH-Px in liver tissue， the protein and mRNA expressions of Nrf2 and HO-1 were significantly lowered in the model group （P＜0.05）； the liver coefficient， the percentage of Masson staining positive area， ALT and AST levels of serum and liver tissue， MDA level of liver tissue， the protein and mRNA expressions of TGF-β1， Smad2， ERK1 and Keap1 were significantly increased （P＜0.05）. Compared with the model group， the liver lesions of rats in each drug group were improved， and the above quantitative indexes were generally reversed （P＜0.05）. CONCLUSIONS Breviscapine has a good intervention effect on HF rats， which may be related to inhibiting TGF-β1/Smad2/ERK1 pathway for anti-fibrosis and regulating Keap1/Nrf2/HO-1 pathway to inhibit oxidative stress.

【Objective】 To understand factors associated with children′s attendance at nursery care institutions (NCIs) and the basic characteristics of the NCIs children were enrolled in, so as to provide scientific evidence for policymakers. 【Methods】 During November 2020 and January 2021, parents who visited the Department of Child Health Care in six selected maternal and children′s hospitals, as well as nearby NCIs in Zhejiang were invited to finish an online questionnaire. Information such as children′s sociodemographic characteristics, parents′ knowledge, attitude and practice regarding nursery and feeding, etc. were collected. A total of 1 756 questionnaires were collected. 【Results】 Compared to children who were not in NCIs, children enrolled in NCIs were older (94.4% of children ≥24 months vs. 30.1%, χ²=835.27), more likely to be from the local area (87.2% vs. 81.4%,χ²=12.25), more likely to have parents with a college degree (mother: 83.6% vs. 74.2%, χ²=35.29; father: 79.9% vs. 70.0%, χ²=27.01), had a higher prevalence of family annual income >200 000 CNY (49.5% vs. 28.2%, χ²=110.49), and were less likely to have their grandparents available to take care of them (16.7% vs. 26.8%, χ²=31.4) The difference all have great significant.(P<0.05). In a multivariate Logistic regression model, the older the child, the more likely they were to attend an NCI (for children aged 6 - 23 months, OR=6.70; for children aged 24 - 35 months, OR=134.03; and for children aged 36 - 42 months, OR=699.33; P<0.05). Family annual income was positively associated with children′s attendance at NCIs (for those earning 100 000 - 200 000 CNY/year, OR=1.63; for those earning 200 000 - 500 000 CNY/year, OR=2.96; and for those earning >500 000 CNY/year, OR=4.62, P<0.05). Conversely, the higher the level of grandparent involvement in daily care, the lower the attendance at NCIs (for children cared for by both parents and grandparents, OR=0.57; for those primarily cared for by grandparents, OR=0.26, P<0.05). For children who used to stay at NCIs, 82.8% stayed at institutions that only recruited children aged 0 - 3 years, 97.4% spent their whole day in NCIs, and 71.4% spent less than 3 000 CNY per month for NCI services. Additionally, over 95% of parents were satisfied with the food and care services in NCIs, as well as their children′s physical development in NCIs. However, 32.1% of NCIs were reported by parents as having no room for breastfeeding. 【Conclusions】 Children′s age, grandparent involvement in routine care, and family annual income are the main factors associated with children′s attendance at NCIs. There is a greater need for more affordable and community-based NCIs, particularly for children under 2 years old. Additionally, more attention should be paid to the quality surveillance, assessment and management of NCIs.

Objective To elucidate the prediction ability of monocyte monolayer assay(MMA)used in hemolytic dis-ease of fetus and newborn(HDFN)caused by IgG anti-M.Methods Plasma from eight pregnant women containing IgG an-ti-M were collected,and were divided into two groups(4 cases with HDFN,with severe clinical symptoms such as fetal hy-drops,and 4 cases without HDFN)according to the clinical outcomes.M antigen positive cells were sensitized with dithioth-reitol(DTT)treated plasma from eight pregnant women respectively.MMA was performed by coincubation with monocytes and sensitized M cells,along with negative and positive control set up.T-test was conducted to compare the difference in phagocytic efficiency between two groups.Results The phagocytic efficiency in group with HDFN were 15.37%,13.05%,9.17%and 24.50%respectively,with the mean value of 15.52%,while the group without HDFN were 8.74%,11.07%,5.12%and 6.23%respectively,with the mean value of 7.79%.There was no significant difference in phagocytic efficiency between two groups(P>0.05).The mean values of both groups were not significantly different from the negative control(P>0.05),but both were significantly lower than positive control(P<0.05).Conclusion The low phagocytic efficiency couldn't convince that the MMA is an effective predictor for the HDFN caused by IgG anti-M,indicating that another mech-anism might be responsible for it rather than monocyte phagocytosis.The assessment of the peak systolic velocity in middle cerebral artery of the fetal should be considered in the management for pregnant women who produce IgG anti-M to estimate the situation of fetal anemia.