Objective To investigate the effect of mutation human proprotein convertase subtilism/kexin type 9(hPCSK9D374Y)in PCSK9 gene on methionine choline deficiency diet(MCD)-induced nonalcoholic steato-hepatitis(NASH)in mice.Methods Sixteen C57BL/6J wild-type mice were selected and randomly divided into the hPCSK9D374Y group and the control GFP group.MCD was fed for 6 weeks,and then the serum level of hepatic triglyceride,alanine aminotransferase(ALT)and aspartate aminotransferase(AST)was examined.Oil Red O and Sirius Red staining microscopy were used to identify hepatic lipid infiltration and fibrosis severity.F4/80-positive cell infiltration was analyzed using immunohistochemistry.Lipid synthesis and inflammatory response-related proteins were detected by Western blot and related mRNA expression was analyzed by RT-qPCR.Results Hepatic hPCSK9 protein and mRNA were significantly up-regulated,LDLR protein expression was down-regulated,and ser-um level of ALT and AST was significantly elevated in the hPCSK9D374Y group of mice(P＜0.05).The degree of he-patic steatosis and fibrosis increased and F4/80-positive cells were significantly increased(P＜0.01).FASN and SCD1 proteins were significantly up-regulated and PPARα was down-regulated in the hPCSK9D374Y group;The ex-pression of TLR4 and p-P65 was elevated,whereas the expression of Iκ Bα was decreased(P＜0.001).RT-qPCR re-sults showed a significant increase of mRNA coding inflammatory factors TNF-α,IL-1β,IL-6,and MCP-1,and a significant up-regulation of fibrosis-associated mRNAs(collagen Ⅰα and collagen Ⅲα)was found(P＜0.001).Conclusions Functionally acquired mutation in the PCSK9 gene(hPCSK9D374Y)exacerbates MCD-induced hepatic steatosis,inflammatory response and fibrosis in mice.

Objective To explore the regulatory mechanism of angiopoietin 1(ANGPT1)on proliferation,invasion and angiogenesis of prostate cancer cells.Methods Mouse prostate cancer cell line(RM-1)was divided into control group,NC-oe group,ANGPT1-oe group,NC-sh group and ANGPT1-sh group.NC-oe,ANGPT1-oe,NC-sh and ANGPT1-sh were transfected into RM-1 cells by Lipofectamine3000 reagent.The transfection efficiency was verified by RT-qPCR and Western blot,cell proliferation was detected by MTT assay and EdU stai-ning and cell invasion was detected by Transwell assay.The cells were divided into the following groups:control group,Tie2-expressing monocytes/macrophages(TEMs)group,NC-oe+TEM group,ANGPT1-oe+TEM group,NC-sh+TEM group and ANGPT1-sh+TEM groups.RM-1 and TEM were co-cultured.RM-1 cells were collected after 72 hours and subjected to MTT proliferation assay,EdU staining assay and Transwell invasion assay.The co-cultured cell supernatant from each group mouse umbilical vein endothelial cells(MUVECs)were co-incubated with cell supernatant collected from each group cells and then to detect the number of tubules formed.The co-cul-tured cell supernatant of each group was collected,and the level of MMP-9,VEGFA and COX-2 were detected by ELISA.Results 1)Compared with control group and NC-oe group,the level of ANGPT1 mRNA and protein in ANGPT1-oe group was increased(P＜0.05).Compared with control group and NC-sh group,the level of ANGPT1 mRNA and protein in ANGPT1-sh group was decreased(P＜0.05).2)Compared with control group,the cell viability,EdU positive rate,counting of invasive cells and of tubules formed in TEM group significantly increased(P＜0.05),the level of MMP-9,VEGFA and COX-2 in the supernatant was increased(P＜0.05).Compared with TEM group and NC-oe+TEM group,the cell viability,EdU positive rate,counting of invasive cells and of tubules formed in ANGPT1-oe+TEM group decreased(P＜0.05).The level of MMP-9,VEGFA and COX-2 in the supernatant was significantly decreased(P＜0.05).Compared with TEM group and NC-sh+TEM group,the cell viability,EdU positive rate,counting of invasive cell and of tubules formed in ANGPT1-sh+TEM group all increased(P＜0.05).The level of MMP-9,VEGFA and COX-2 in the supernatant were increased(P＜0.05).Conclusions The decreased expression of ANGPT1 in prostate cancer significantly enhances the promotion effect of TEMs on the proliferation,invasion and angiogenesis of prostate cancer cells,thus promoting the progression of prostate cancer.

Objective To investigate the mechanism of reactive oxygen species(ROS)and pyroptosis in amioda-rone-induced lung epithelial cell injury.Methods The viability of cells treated with amiodarone,amiodarone+N-acetylcysteine(ROS inhibitor,NAC)and amiodarone+NF-κB inhibitors was detected by MTT assay.ROS level was detected by DCFA probe and the levels of IL-6,IL-18,IL-1β and TNF-α in the supernatant were detected by ELISA;NLRP3.Caspase-1 p20 and GSDMD-N protein were detected by Western blot.Results Amiodarone con-centration-dependently reduced cell viability(P＜0.05),NAC and NF-κB attenuated the cytotoxicity of amiodarone(P＜0.05).Amiodarone increased levels of reactive oxygen species,inflammatory factors and pyroptosis-related proteins in lung epithelial cells(P＜0.05),while NAC and NF-κB inhibitors reduced the levels of these indicators(P＜0.05).Conclusions NF-κB inhibitor or NAC attenuates amiodarone-induced injury of human lung epithelial cell line in vitro.

Objective OCTA analysis was employed to assess the alterations in retinal microcirculation following vitrectomy in patients with diabetic retinopathy.Methods The clinical data of 60 eyes from 60 diabetic retinopathy patients who accepted vitrectomy from Mar.2022 to May.2024 in Daqing Oilfield General Hospital were analyzed prospectively.The preoperative and postoperative outcomes of best corrected visual acuity(BCVA),intraocular pres-sure(IOP),and optical coherence tomography angiography(OCTA)were compared at baseline,as well as at 1 day,1 week,1 month,and 3 months following surgery.Results There was statistically significant differences in IOP between preoperative and postoperative 1 day and 1 week(P＜0.01).There were significant differences in BCVA and CMT observed prior to surgery,as well as at 1 day,1 week,1 month,and 3 months post-surgery(P＜0.01).FAZ and RPC exhibited significant differences prior to surgery,as well as at 1 week,1 month,and 3 months postoperatively(P＜0.01).The SVD,DVD,and RNFL exhibited significant differences prior to surgery and at 1 month and 3 months postoperatively(P＜0.01).Conclusions Vitrectomy can substantially enhance the visual acuity of patients with DR,and this improvement tends to stabilize approximately one month post-surgery,potential-ly correlating with the stability of central macular thickness(CMT)observed in patients at that time.The procedure can significantly decrease intraocular pressure in patients,and FAZ along with the radial peripapillary capillaries(RPC)in the macular region exhibited earlier improvement postoperatively.

Objective To investigate the impact of DNA damage-regulated autophagy factor 2(DRAM2)on the proliferation and migration of non-small cell lung cancer(NSCLC)cells through tumor protein p53(p53)and autophagy.Methods DRAM2 gene was knocked down using lentiviral technology to establish NSCLC cell lines,and autophagy markers were detected by immunofluorescence and Western blot.CCK8 and Transwell assays were used to detect cell proliferation and migration.The effects of autophagy activation and p53 knockdown on autoph-agy and functions of DRAM2?knockdown cells were examined.Results Knockdown of DRAM2 inhibited NSCLC cells,with upregulation of p62 expression(P<0.05)and decreased level of LC3?Ⅱ(P<0.05).Knockdown of DRAM2 suppressed the proliferation(P<0.001)and migration(P<0.001)of NSCLC cells.Activation of auto?phagy partially reversed the inhibitory effects of DRAM2 knockdown on cell proliferation(P<0.01)and migration(P<0.01).When DRAM2 and p53 were knocked down simultaneously,autophagy,cell proliferation(P<0.05)and migration abilities(P<0.001)were restored.Conclusions Knockdown of DRAM2 inhibits the proliferation and migration of lung cancer cell line A549,providing a potential intervention direction for the devel?opment of therapeutic strategies.

Objective To explore the potential impact of unhealthy diets on cardiovascular diseases and all-cause mortality.Methods This study included the individuals aged 35-70 years from 45 cities and 70 rural communities across 12 provinces in China,as part of the Prospective Urban Rural Epidemiology(PURE)study.Dietary habits were assessed using a food frequency questionnaire.The dietary health status was scored using the Alternative Healthy Eating Index(AHEI),with participants in the lowest tertile of AHEI being categorized into the unhealthy diet group,while others were classified as the healthy diet group.The primary endpoints included major cardiovas-cular diseases(myocardial infarction,stroke,or heart failure)and all-cause mortality.Cox proportional hazard models were used to estimate hazard ratios(HR)for each group.Results A total of 40 925 participants were in-cluded in the study,with a median follow-up time of 11.9 years(interquartile range 9.6-12.6 years).During this period,2 066 deaths and 3 099 cases of major cardiovascular diseases were reported.The results showed that un-healthy diet increased the risk of major cardiovascular diseases by 10%(HR=1.10,95%CI:1.02-1.20,P＜0.05)and all-cause mortality by 7%(HR=1.07,95%CI:1.00-1.18,P＜0.05).Among male residents,un-healthy diet did not increase the risk of major cardiovascular diseases or all-cause mortality.However,among female residents,those with an unhealthy diet had a higher risk of major cardiovascular diseases(HR=1.12,95%CI:1.00-1.25,P＜0.05)and all-cause mortality(HR=1.26,95%CI:1.08-1.46,P＜0.05)compared to those with a healthy diet.Conclusions Unhealthy diet increases the risk of major cardiovascular diseases and all-cause mortality,particularly among women.There is a need to raise awareness about healthy dietary to prevent death and the occurrence of major cardiovascular diseases.

Objective To reveal the thermally induced denaturation of wild-type human γ D-crystallin(HGD)and congenital cataract-related mutant(HGD P23T),and compare the differences in the structural changes between wild-type and mutants during a heating process.Methods HGD and HGD P23T were expressed and purified.The temperature-dependent intrinsic fluorescence intensity and static light scattering intensity of the protein samples were measured to reveal the temperature-dependent folding and aggregation structural changes of HGD and HGD P23T.Results When the temperature was below 70℃,the barycentric mean of the intrinsic fluorescence of HGD and HGD P23T shifted towards a longer wavelength with increasing temperature and the fluorescence intensity de-creased indicating the unfolded protein conformations.The conformational stability of HGD P23T was weaker than that of HGD.When temperature was higher than 70℃,the static light scattering intensity increased significantly with temperature,indicating protein aggregation upon heating.Relative to the wild-type,HGD P23T showed a stronger aggregation potency.Conclusions Heating disrupts the folding conformation of Γd-crystallin,induces the unfolded protein to aggregate.The disease-associated P23T mutation significantly reduces the conformational stability of Γd-crystallin.

Objective To investigate the effect and mechanism of melatonin on the anoikis of melanoma cells.Methods The drug concentration of melatonin inhibiting melanoma cell line B16-F10 was optimized based on the effect on CCK-8 assay.An anti-anoikis of melanoma cell model was developed and divided it into four groups:The blank control group,the TrkB activator group,the melatonin group and the melatonin+TrkB activator group.Calce-in AM/EthD-1 fluorescence double staining was used to detect the anoikis of melanoma cells.Reactive oxygen spe-cies were detected using the fluorescent probe DCFH-DA.Western blot was used to detect the expression of Nrf2 protein and TrkB protein in each group.Results Melatonin significantly inhibited the proliferation of melanoma cells in a time-and dose-dependent manner with IC50 of 1×10-7 μmol/L.Its inhibitory effect was found to be related to in-duction of anoikis of melanoma cells.Melatonin could upregulate the generation of cellular reactive oxygen species(P<0.05),while addition of TrkB activator antagonized this effect.Melatonin could reduce the expression of Nrf2 protein and TrkB protein in melanoma cells(P<0.05),and the addition of TrkB activator could inhibite the effect of melatonin on the expression of Nrf2 protein and TrkB protein(P<0.05).Conclusions Melatonin can inhibit the pro-liferation of melanoma cell line B16-F10 through the mechanism of inducing anoikis.

Objective To explore the impact of change in oxygen concentration on the expression of angiopoietin-like protein 8(ANGPTL8)by endothelial cells(HPAECs)of human pulmonary artery,and the role and mecha-nism of ANGPTL8 in pulmonary hypertension(PH).Methods HPAECs were treated under hypoxic and hyperoxic conditions,and the expression level of ANGPTL8 was detected using Western blot and PCR.The changes in endo-thelial-mesenchymal transition(EndMT)and ERK signaling pathway activity were analyzed.Simultaneously,new-born rats were exposed to hyperoxia to develop a bronchopulmonary dysplasia(BPD)model.The expression of ANGPTL8 protein and changes in the ERK signaling pathway in lung tissue were observed.Results Under hypoxic condition,the protein expression of ANGPTL8 in HPAECs was significantly increased accompanied by inhibition of the ERK signaling pathway.ANGPTL8 promoted the EndMT process induced by hypoxia(P<0.05)and silencing the expression of ANGPTL8 resulted in a partial reversal of EndMT.The protein expression of ANGPTL8 was decreased in hyperoxia-exposed HPAECs and rat lung tissues accompanied by the activation of the ERK signaling pathway(P<0.05).Conclusions ANGPTL8 is highly sensitive to the change of oxygen concentration in HPAECs and closely correlated to its expression level and to the activity of ERK signaling pathway.This result suggests that ANGPTL8 may have potential regulatory effects on the development of PH.

Objective To investigate the effects of CD151 down-regulation combined with bevacizumab on colorectal cancer growth and microvessel density.Methods Human colorectal cancer cell line HT-29 and CD151--HT-29 cells strain(CD151 down-regulated HT-29 cells)were treated with bevacizumab.The cells were divided into four groups:control(HT-29)group,bevacizumab-treatment group,CD151--HT-29 group,and CD151--HT-29+bevacizumab-treatment group.Cell proliferation was observed in each group using the MTS assay.A subcutaneous xenograft model in nude mice was established,and the HT-29 control group and CD151--HT-29 group were treated with either 0.9％NaCl solution or bevacizumab.The growth of subcutaneous tumors in the four groups was ob-served,and the volume and weight of the tumors were recorded.Tumor tissues were collected for immunohistochem-ical staining of endothelial cells to assess microvessel density(MVD).Results Compared with the control group,cell proliferation was significantly reduced in the bevacizumab-treated group and CD151--HT-29 group(P<0.001).Cell proliferation in the CD151--HT-29+bevacizumab-treated group was slower than that in the single treatment groups(P<0.001).In the subcutaneous tumor model,the volume,weight,and MVD of tumors in the bevacizumab-treated and CD151--HT-29 groups were significantly reduced compared to the control group(P<0.01).In the CD151--HT-29+bevacizumab group,the tumor volume,weight,and CD34 expression were significantly lower than in the single treatment groups(P<0.01).Conclusions CD151 protein may play a role in the regulation of angiogenesis in colorectal cancer tissues and may have a synergistic effect with bevacizumab in in-hibiting microvessel formation in tumor tissues.