Objective : To investigate the effect of miR-141-3p on LPS induced A549 cell injury by targeting high mobility group protein 1 (HMGB1) ． Methods : A549 cells derived from type Ⅱ alveolar epithelial cells were taken as the study object，miR-141-3p mimics，mimics NC，HMGB1 gene overexpression plasmid (pcDNA3. 1-HMGB1) and empty Vector were transfected into A549 cells respectively or co-transfected，then 10 μg / ml LPS was used for 24 h．Cell proliferation activity was detected by cell counting kit-8 ( CCK-8) ．The activity of lactate dehydrogenase ( LDH) in the supernatant of cell culture was detected by colorimetry．The apoptosis level of each group was detec- ted by flow cytometry．The levels of interleukin (IL) -1 β , IL-6 and tumor necrosis factor α (TNF-α) were detected by enzyme-linked immunosorbent assay (Elisa) ．Dual luciferase reporter gene assay verified the targeted regulatory relationship between miR-141-3p and HMGB1 . Results : After treatment with LPS ，the proliferative activity of A549 cells and the expression level of miR-141-3p decreased ( P ＜0. 05 ) ，the apoptosis rate increased ( P < 0. 05) ，the levels of IL-1 β , IL-6，TNF-α and the activity of LDH in supernatant increased (P＜0. 05) ．Overex- pression of miR-141-3p increased the proliferation activity of A549 cells treated with LPS (P ＜0. 05 ) ，and de- creased the apoptosis rate and the levels of IL-1 β , IL-6，TNF-α in cells and LDH activity in supernatant (P < 0. 05) ．However，overexpression of HMGB1 gene could reverse the ameliorative effect of miR-141-3p on LPS-in- duced A549 cell injury．Dual luciferase reporter gene experiment confirmed that HMGB1 was the downstream target gene of miR-141-3p． Conclusion miR-141-3p can inhibit LPS-induced apoptosis，reduce the expression level of inflammatory factors，and improve the damage of A549 cells，which may be related to the targeted regulation of HMGB1 expression.

Anaplastic thyroid carcinoma(ATC)is the most aggressive malignancy with poor prognosis.The pathogenesis of ATC is complex,and there is no effective treatment at present.In recent years,with the deep understanding of the genetic(such as BRAF V600E,TP53,TERT,PIK3CA mutations,etc.)and epigenetic(such as histone methylation,histone deacetylation,microRNA regulatory pathways,etc.)changes driving ATC,molecular targeted therapy has brought new hope to ATC patients.This article reviews the molecular mechanisms of ATC and the latest achievements in targeted therapy and other therapies.

Objective:To investigate the expression of HBB in anaplastic thyroid cancer(ATC)cells and its regulatory effect on proliferation,invasion,migration and apoptosis of ATC cells and its potential mechanism.Methods:The expression of HBB in thyroid cancer and paracancerous tissues was analyzed through TIMER database.The correlation between the expression of HBB and the overall survival time of thyroid cancer patients was analyzed through KM-plotter database.The expression of HBB mRNA in ATC cells was detected by RT-qPCR.The HBB knockout or overexpression plasmid was transfected into ATC cells.The expression of HBB protein was detected by Western blot.The proliferation activity was detected by CCK-8 assay.The migration and invasion ability was detected by Transwell assay.The apoptosis was detected by flow cytometry.The expression of β-catenin was detected by Western blot.Results:The expression of HBB was low in thyroid cancer,and the overall survival time of patients with high expression of HBB was high.The expression of HBB protein was down-regulated in ATC cells.Knockout of HBB increased the ability of proliferation,migration and invasion of ATC cells and the expression of β-catenin protein,and inhibited apoptosis.However,overexpression of HBB decreased the ability of proliferation,migration and invasion of ATC cells and the expression of β-catenin protein,and promoted apoptosis.Conclusions:High HBB expression is associated with higher overall survival in patients with thyroid cancer.It may inhibit the proliferation,migration and invasion of ATC cells and promote apoptosis through Wnt/β-catenin signal pathway.

Objective:To discuss the effect of inhibiting microRNA(miR)-193a-5p expression on pulmonary fibrosis in the rats with acute respiratory distress syndrome(ARDS),and to clarify the related mechanism.Methods:A total of 60 male SD rats were divided into sham operation group,model group,miR-193a-5p antagonist group(Antagomir group),and negative control group(Antagomir-NC group),and there were 15 rats in each group.The ARDS animal model was induced by administering 10 mg·kg-1 lipopolysaccharide(LPS)via tracheal instillation,while the rats in sham operation group received an equal volume of saline.After successful modeling,the rats in Antagomir group and Antagomir-NC group were treated with miR-193a-5p Antagomir or Antagomir-NC via tail vein injection.The arterial partial pressure of oxygen(PaO2)and oxygenation index(OI)of the rats in various groups were measured;HE staining and Masson staining were used to observe the pathology and collagen fiber deposition in lung tissue of the rats;kit was used to detect the level of hydroxyproline(Hyp)in lung tissue of the rats in various groups;enzyme-linked immunosorbent assay(ELISA)method was used to detect the levels of inflammatory factors tumor necrosis factor α(TNF-α),interleukin(IL)-1β,and IL-6 in bronchoalveolar lavage fluid(BALF)of the rats in various groups;real-time fluorescence quantitative PCR(RT-qPCR)method was used to detect the expression levels of miR-193a-5p in lung tissue of the rats in various groups;Western blotting method was used to detect the expression levels of β-catenin,Snail family transcriptional repressor 1(Snail1),and α-smooth muscle actin(α-SMA)proteins in lung tissue of the rats in various groups.Results:Compared with sham operation group,the PaO2 and OI of the rats in model group were significantly decreased(P<0.05);compared with model group,the PaO2 and OI of the rats in Antagomir group were significantly increased(P<0.05).The HE staining results showed that the lung tissue structure of the rats in sham operation group was normal,and there were no obvious inflammatory changes;compared with sham operation group,mild abnormalities in lung tissue structure,alveolar atrophy,and collapse were observed in the rats in model group and Antagomir-NC group,with a large number of lymphocytes and a small number of neutrophils infiltrating in the alveolar cavities,and widened alveolar spaces;compared with model group,the rats in Antagomir group showed a significant reduction in lymphocytes and neutrophil infiltration in the alveolar cavities and there were no obvious hyperplasia.The Masson staining results showed no obvious blue collagen fiber deposition in lung tissue of the rats in sham operation group;compared with sham operation group,significant blue collagen fiber deposition was observed in lung tissue of the rats in model group and Antagomir-NC group,with severe damage of the alveolar structure,indicating obvious pulmonary fibrosis;compared with model group,the deposition of blue-stained collagen fibers in lung tissue of the rats in Antagomir group was significantly reduced.Compared with sham operation group,the level of Hyp in lung tissue of the rats in model group was significantly increased(P<0.05);compared with model group,the level of Hyp of the rats in Antagomir group was significantly decreased(P<0.05).The ELISA results showed that compared with sham operation group,the levels of TNF-α,IL-1β,and IL-6 in BALF of the rats in model group were significantly increased(P<0.05);compared with model group,the levels of TNF-α,IL-1β,and IL-6 of the rats in Antagomir group were significantly decreased(P<0.05).The RT-qPCR results showed that compared with sham operation group,the expression level of miR-193a-5p in lung tissue of the rats in model group was significantly increased(P<0.05);compared with model group,the expression level of miR-193a-5p of the rats in Antagomir group was significantly decreased(P<0.05).The Western blotting results showed that compared with sham operation group,the expression levels of β-catenin,Snail1,and α-SMA proteins in lung tissue of the rats in model group were significantly increased(P<0.05);compared with model group,the expression levels of β-catenin,Snail1,and α-SMA proteins in lung tissue of the rats in Antagomir group were significantly decreased(P<0.05).Conclusion:Inhibition of miR-193a-5p expression can improve the lung function and alleviate the pulmonary fibrosis in the ARDS rats by reducing the inflammatory responses and downregulating the expressions of β-catenin,Snail1,and α-SMA proteins.

Objective To explore the effects of the long non-coding RNA(lncRNA)NK2 homeobox 1-antisense RNA 1(NK2-1-AS1),which mediates the microRNA(miR)-96-5p/PR domain-containing protein 16(PRDM16)axis,on anaplastic thyroid cancer(ATC)cell proliferation,migration,and invasion in vitro and transplanted tumor growth in vivo.Methods The differentially expressed lncRNA NKX2-1-AS1 in ATC tissues and cells,its target miRNA miR-96-5p,and its downstream target gene PRDM16 were screened using a bioinformatics analysis.The dual-luciferase reporter assay validated the relationship between NKX2-1-AS1 and miR-96-5p as well as the connection between miR-96-5p and PRDM16.Western blotting was performed to detect the effect of miR-96-5p overexpression on PRDM16 in CAL-62 cells overexpressed with NKX2-1-AS1.Plate clone formation,scratch,and Transwell assays were used to detect the effects of PRDM16knockdown on the proliferation,migration,and invasion of CAL-62 cells overexpressing NKX2-1-AS1.CAL-62 cells were injected subcutaneously into nude mice and the effect was observed of PRDM16knockdown on the growth of transplanted tumors of CAL-62 cells overexpressing NKX2-1-AS1.Results The bioinformatics analysis revealed that the NK2-1-AS1/miR-96-5p/PRDM16 axis was involved in regulating ATC development.The dual-luciferase reporter assay demonstrated that NKX2-1-AS1 bound to miR-96-5p and miR-96-5p bound to PRDM16.NKX2-1-AS1 overexpression upregulated PRDM16 protein expression in CAL-62 cells,while miR-96-5p overexpression reversed this phenomenon.NKX2-1-AS1 overexpression inhibited CAL-62 cellular proliferation,migration,and invasion in vitro and transplanted tumor growth in vivo,while knocking down PRDM16 reversed these phenomena.Conclusion NK2-1-AS1 may compete with miR-96-5p as an endogenous RNA to bind to its downstream target gene,PRDM16,and upregulate its expression,thus inhibiting ATC cell proliferation,migration,and invasion in vitro and transplanted tumor growth in vivo.

Objective To explore the effect of transcription factor E2F7 on the proliferation,migration,invasion,and tumor growth of anaplastic thyroid cancer(ATC)cells in vitro and to elucidate the underlying mechanisms.Methods Lentivirus transfection was used for a stable E2F7 knockdown in CAL-62 cells,and real-time PCR was used to detect E2F7 expression in these cells to verify the trans-fection efficiency.CAL-62 cells were divided into sh-NC and sh-E2F7 groups,and cell proliferation was measured using the CCK-8 assay,whereas cell migration and invasion abilities were measured using the Transwell assay.CAL-62 cells were subcutaneously injected into nude mice to observe tumor growth.The EPD website predicted an E2F7 binding site on the CXCL5 promoter,and the dual-lucif-erase reporter gene assay measured the effect of E2F7 knockdown on the luciferase activity of the CXCL5 promoter.The impact of E2F7 knockdown on CXCL5 levels in CAL-62 cells was assessed through real-time PCR and ELISA.Further,CAL-62 cells were divided into sh-E2F7+vector and sh-E2F7+CXCL5 groups to study the effects of CXCL5 overexpression on cell proliferation,migration,invasion,and the CXCR2/ERK signaling pathway following E2F7 knockdown.Results E2F7 knockdown inhibited CAL-62 cell proliferation,migra-tion,and invasion in vitro and tumor growth in vivo.The CXCL5 promoter has an E2F7 binding site,and E2F7 knockdown reduced the luciferase activity of the CXCL5 promoter.CXCL5 overexpression reversed the inhibitory effect of E2F7 knockdown on cell proliferation,migration,invasion,and the CXCR2/ERK signaling pathway in CAL-62 cells.Conclusion E2F7 promotes ATC cell proliferation,migra-tion,invasion,and tumor growth in vitro by activating the CXCL5/CXCR2/ERK signaling pathway mediated by CXCL5 transcription.

OBJECTIVE To investigate the modulating effect of neotropics on form deprivation myopia(FDM)in guinea pigs.METHODS Tricolour guinea pigs were randomly divided into normal control group,FDM model group,FDM+saline group,FDM+atropine group,and FDM+neotropine group,with eight animals in each group.Except for the normal control group,the right eyes of the guinea pigs were covered for 14 d to establish a guinea pig FDM model.The drug administration groups were injected with 10 μL of saline,1%atropine,or 1%neotropine into the vitreous cavity once every other day.The changes in the refractive error and axial length of both eyes were recorded for 1 d before the intervention and for 14 d after the intervention.Then,the eyeballs of guinea pigs were taken from the right eyes.HE staining was used to evaluate the histopathological structure of the sclera while sirius red staining was used to detect the collagen protein content in the sclera.RT-qPCR was used to detect the mRNA expressions of transforming growth factor-β1(TGF-β1),matrix metalloproteinase(MMP-2)and tissue inhibitor of metalloproteinase(TIMP-2)in guinea pigs'sclera.The protein expression levels of collagen type Ⅰ(Col-Ⅰ)and TGF-β1 in guinea pig sclera were detected by Western blotting while those of MMP-2,TIMP-2 and Ki-67 were detected by immunohistochemical staining.RESULTS Compared with the nor-mal control group,eyes of guinea pig in the FDM model group showed a significantly lower refractive error(P<0.01),significant elongation of the ocular axis(P<0.01),scattered distribution of scleral fibre bundles,sparse collagen cells,reduced scleral thickness(P<0.01),and a significantly lower collagen protein content(P<0.01).The mRNA and protein expressions of TIMP-2 and TGF-β1 were lower(P<0.05,P<0.01),and MMP-2 was higher(P<0.01)in scleral tissue.The protein expression level of Col-Ⅰwas lower(P<0.05)while that of Ki-67 was elevated(P<0.01)in scleral tissue.Compared with the FDM model group,there were no significant changes in any of the indexes in the FDM+saline group.The refractive error of the right eyes of guinea pigs in the FDM+neotropium group and the FDM+atropium group were significantly higher(P<0.05),the length of the ocular axis was significantly shorter(P<0.05),the collagen fibres were arranged more tightly,the fibre bundles were distributed more orderly,the distribution of the collagen cells was more uniform,and the thickness of the sclera was significantly increased(P<0.01).Collagen protein contents were significantly higher(P<0.05,P<0.01),the mRNA and protein expressions of TIMP-2 and TGF-β1 were significantly higher(P<0.01),MMP-2 mRNA and protein expressions were significantly lower(P<0.05,P<0.01).The protein expression level of Col-Ⅰwas higher(P<0.05,P<0.01),and that of Ki-67 was lower(P<0.05,P<0.01)in scleral tissue.CONCLU-SION The muscarinic antagonist neotropine inhibits the development of myopia in guinea pigs in the FDM model by reversing both the down-regulation of TGF-β1 and the up-regulation of MMP-2 in scleral tissues and inhibiting the remodeling of the scleral extracellular matrix.

Objective To investigate the effect and mechanism of miR-141-3p on pulmonary fibrosis in rats with acute respiratory distress syndrome(ARDS).Methods Rats were divided into the control group,the model group,the agomir negative control group and the miR-141-3p agomir group according to random number table,with 10 rats in each group.In addition to the control group,the ARDS rat model was established by lipopolysaccharide(LPS)infusion.Rat alveolar typeⅡepithelial cells RLE-6TN cells were divided into the NC group,the LPS group,the miR-NC group,the miR-141-3p mimics group,the miR-141-3p mimics+pcDNA group and the miR-141-3p mimics+NRF2 and Kelch-like ring associated protein 1(Keap1)group.LPS cell model was established in all groups except the NC group.The mRNA expression levels of miR-141-3p and Keap1 in lung tissue and cells were detected by qPCR.Western blot assay was used to analyze lung tissue and cell epithelial cadherin(E-cadherin),neural cadherin(N-cadherin),microtubule associated protein light chain 3B(LC3B),autophagy associated gene Beclin-1,α-smooth muscle actin(α-SMA),type I collagen(Col-Ⅰ),Keap1 and nuclear factors E2 related factor 2(NRF2)and heme oxygenase 1(HO-1).HE staining and Masson staining were used to observe pathological changes of lung tissue and to estimate the area of lung tissue injury and pulmonary fibrosis.Hydroxyproline(Hyp)in lung tissue was detected by the kit.Levels of inflammatory factor interleukin-1β,tumor necrosis factor(TNF-α)and oxidative stress index malondialdehyde(MDA)and superoxide dismutase(SOD)were detected by ELISA.Dual luciferase reporting experiment was used to verify the targeting relationship between miR-141-3p and Keap1.Results The expression of miR-141-3p was down-regulated and the expression of Keap1 was up-regulated in lung tissue and cells(P＜0.05).Overexpression of miR-141-3p can reduce the degree of pathological damage and fibrosis of lung tissue in rats,Hyp content,and up-regulate expression levels of SOD,E-cadherin,LC3B,Beclin-1,NRF2 and HO-1 in lung tissue and cells,and down-regulate the expression levels of IL-1β,TNF-α,MDA,N-cadherin,α-SMA,Col-I and Keap1(P＜0.05).Overexpression of Keap1 was able to reverse the improvement effect of overexpression of miR-141-3p on alveolar epithelial cell damage in ARDS rats(P＜0.05).Double Luciferase reporter gene experiment confirmed that miR-141-3p and Keap1 may have a targeted regulatory relationship.Conclusion Overexpression of miR-141-3p may activate the Keap1-NRF2/ARE signaling pathway,activate autophagy,inhibit inflammatory response,oxidative stress,and EMT progression,and improve pulmonary fibrosis in ARDS rats.

Objective:To investigate the value of evaluating pancreatic exocrine insufficiency with fecal pancreatic elastase-1 in the clinical staging of chronic pancreatitis and prognosis evaluation.Methods:A total of 100 patients with pancreatic exocrine insufficiency (patient group) who received treatment in Wenzhou Central Hospital from January 2021 to June 2022 and 100 subjects without pancreatic exocrine insufficiency (control group) were included in this study. Fecal pancreatic elastase-1 content was measured by an enzyme linked immunosorbent assay. The receiver operating characteristic (ROC) curve was plotted to evaluate the value of fecal pancreatic elastase-1 content in the diagnosis of pancreatic exocrine insufficiency. Fecal pancreatic elastase-1 content was compared among patients with different clinical stages of chronic pancreatitis. The factors that affect the prognosis of patients with chronic pancreatitis were analyzed using logistic regression analysis.Results:Pancreatic elastase-1 content in the patient group was (63.28 ± 13.24) μg/g, which was significantly lower than (768.29 ± 102.59) μg/g in the control group ( t = 68.16, P < 0.05). The sensitivity, specificity, Youden index, and 95% CI of using pancreatic elastase-1 content to diagnose pancreatic exocrine insufficiency were 74.7%, 63.5%, 0.724, and 0.740-0.870, respectively. Among the 200 included subjects, 103 had chronic pancreatitis. With the increase in M-ANNHEIM clinical stage, fecal pancreatic elastase-1 content in patients with chronic pancreatitis gradually decreased ( F = 182.66, P < 0.05). Pancreatic elastase-1 content < 200 μg/g was used as a standard to evaluate pancreatic exocrine function. Results showed that 35 patients had stage I chronic pancreatitis, 40 patients had stage II chronic pancreatitis, and 28 patients had stage III chronic pancreatitis. There was no significant difference in the number of patients with different stages of chronic pancreatitis between the two clinical stage classification methods ( χ2 = 12.46, P = 0.002). Six-month follow-up results showed that among 103 patients with chronic pancreatitis, 31 had a poor prognosis (30.1%). Univariate analysis revealed that there were significant differences in age at onset, body mass index, triglyceride level, alcohol consumption, and pancreatic elastase-1 content among patients with different prognoses ( χ2 = 24.07, 4.27, 5.43, 8.38, 4.93, P < 0.05). Multivariate logistic regression analysis showed that age at onset, body mass index, triglyceride level, alcohol consumption, and pancreatic elastase-1 content were the independent influential factors of prognosis in patients with chronic pancreatitis [ OR (95% CI) = 4.207 (2.741-11.609), 1.870 (1.241-2.972), 1.984 (1.437-3.113), 2.769 (1.827-5.125), 1.951 (1.469-3.387), all P < 0.05]. Conclusion:Pancreatic elastase-1 content is of great value in assessing pancreatic exocrine insufficiency, and is closely related to the clinical staging and prognosis of patients with chronic pancreatitis. Therefore, fecal pancreatic elastase-1 content is expected to be a reliable reference for assessing the progress of chronic pancreatitis and predicting its prognosis.

Objective:To explore the research status and trends of sarcopenic obesity at home and abroad, so as to provide a reference for the nursing staff to carry out related clinical work or research.Methods:The articles on sarcopenic obesity included from January 1, 2000 to December 31, 2021 were retrieved in the Web of Science core database. Bibliometric analysis was performed using CiteSpace software.Results:From 2000 to 2021, there were 1 252 articles related to sarcopenic obesity, and the number of articles published showed an upward trend year by year. Totals of 83 countries or regions participated in the research, and the country with the largest number of publications was the United States (286) . The most published journal was Clinical Nutrition (44) . The author with the most papers was Wolfgang Kemmler (14) , and the institution with the most papers was Korea University (57) . Research hotspots mainly focused on immune resistance, disease risk and nutritional intervention.Conclusions:Bibliometric analysis reveals research patterns and trends in sarcopenic obesity. Foreign countries started early in this research field, and my country is still in its infancy. There is also an urgent need to cultivate research leaders in the field of sarcopenic obesity, form an inter-institutional cooperation mechanism, and carry out multidisciplinary teamwork to promote the health of the elderly.