Objective·To investigate the role of caspase recruitment domain-containing protein 9(CARD9)in regulating macrophage polarization in a rat model of severe acute pancreatitis(SAP).Methods·SD rats were divided into 4 groups:Control group,SAP group,SAP+CARD9 shRNA group,and SAP+Control shRNA group with six rats in each group.The SAP rats transfected with CARD9 shRNA were established by injecting CARD9 shRNA adenovirus 48 hours before the SAP model was induced.The pancreatic tissues,peripheral blood,and peritoneal macrophages were collected 12 hours after the model was established.The expressions of CARD9 gene and CARD9 protein in peritoneal macrophages and pancreatic tissues were measured by real-time PCR and Western blotting.The expressions of TNF-α,IL-6,IL-10 and Arg-1 mRNA were detected by real-time PCR and the polarization types of peritoneal macrophages were detected by flow cytometry.Results·The expressions of CARD9 gene and CARD9 protein in peritoneal macrophages in CARD9 shRNA rats were significantly lower than those in SAP rats and interference control rats,which confirmed the success of CARD9 interference model.Compared with SAP rats,CARD9 shRNA rats had significantly reduced degree of inflammation and pathological scores;the mRNA levels of TNF-α,and IL-6 in peritoneal macrophages were significantly decreased;meanwhile,the mRNA levels of IL-10 and Arg-1 were increased,and the changes in TNF-α and IL-6 were significantly higher than those of IL-10 and Arg-1.The proportion of M1 macrophages was significantly reduced,and the ratio of M1/M2 was significantly decreased.The expression level of CARD9 mRNA in peritoneal macrophages was positively correlated with the proportion of M1 macrophages and the mRNA levels of TNF-α and IL-6.Conclusion·CARD9 is involved in regulating macrophage polarization in SAP rats,and it mainly regulates M1 polarization.Inhibition of CARD9 expression can reduce M1 macrophage polarization and reduce the inflammatory response in SAP rats.

Cardiovascular disease(CVD)is a leading cause of death worldwide,posing a substantial burden on so-ciety.Transfer RNA-derived fragment(tRF)is a novel class of non-coding small RNA produced primarily through the specific cleavage of precursor or mature transfer RNA(tRNA).Unlike the canonical role of tRNA in amino acid trans-port,tRF plays important roles in regulating gene transcription,translation,and epigenetic modifications,thereby influen-cing pathophysiological processes such as cell proliferation,differentiation,and apoptosis.In recent years,with the wide-spread application of small RNA sequencing technologies,a growing number of studies have demonstrated close associations between tRF and the development and progression of various CVDs,making them a research hotspot in the field.This re-view systematically summarizes the origin,classification,functional mechanisms,and recent advances in tRF research re-lated to CVD,aiming to provide new directions for understanding the pathogenesis of CVD and identifying potential thera-peutic targets.

Cardiovascular disease(CVD)is a leading cause of death worldwide,posing a substantial burden on so-ciety.Transfer RNA-derived fragment(tRF)is a novel class of non-coding small RNA produced primarily through the specific cleavage of precursor or mature transfer RNA(tRNA).Unlike the canonical role of tRNA in amino acid trans-port,tRF plays important roles in regulating gene transcription,translation,and epigenetic modifications,thereby influen-cing pathophysiological processes such as cell proliferation,differentiation,and apoptosis.In recent years,with the wide-spread application of small RNA sequencing technologies,a growing number of studies have demonstrated close associations between tRF and the development and progression of various CVDs,making them a research hotspot in the field.This re-view systematically summarizes the origin,classification,functional mechanisms,and recent advances in tRF research re-lated to CVD,aiming to provide new directions for understanding the pathogenesis of CVD and identifying potential thera-peutic targets.

Objective·To investigate the role of caspase recruitment domain-containing protein 9(CARD9)in regulating macrophage polarization in a rat model of severe acute pancreatitis(SAP).Methods·SD rats were divided into 4 groups:Control group,SAP group,SAP+CARD9 shRNA group,and SAP+Control shRNA group with six rats in each group.The SAP rats transfected with CARD9 shRNA were established by injecting CARD9 shRNA adenovirus 48 hours before the SAP model was induced.The pancreatic tissues,peripheral blood,and peritoneal macrophages were collected 12 hours after the model was established.The expressions of CARD9 gene and CARD9 protein in peritoneal macrophages and pancreatic tissues were measured by real-time PCR and Western blotting.The expressions of TNF-α,IL-6,IL-10 and Arg-1 mRNA were detected by real-time PCR and the polarization types of peritoneal macrophages were detected by flow cytometry.Results·The expressions of CARD9 gene and CARD9 protein in peritoneal macrophages in CARD9 shRNA rats were significantly lower than those in SAP rats and interference control rats,which confirmed the success of CARD9 interference model.Compared with SAP rats,CARD9 shRNA rats had significantly reduced degree of inflammation and pathological scores;the mRNA levels of TNF-α,and IL-6 in peritoneal macrophages were significantly decreased;meanwhile,the mRNA levels of IL-10 and Arg-1 were increased,and the changes in TNF-α and IL-6 were significantly higher than those of IL-10 and Arg-1.The proportion of M1 macrophages was significantly reduced,and the ratio of M1/M2 was significantly decreased.The expression level of CARD9 mRNA in peritoneal macrophages was positively correlated with the proportion of M1 macrophages and the mRNA levels of TNF-α and IL-6.Conclusion·CARD9 is involved in regulating macrophage polarization in SAP rats,and it mainly regulates M1 polarization.Inhibition of CARD9 expression can reduce M1 macrophage polarization and reduce the inflammatory response in SAP rats.

Objective To investigate the effect of vitamin D supplementation in early pregnancy on the incidence and glucose metab-olism of high-risk women with gestational diabetes mellitus.Methods Pregnant women who had regular prenatal examination in Hefei First People's Hospital from December 2019 to May 2022 with serum 25(OH)D level＜30ng/ml,expected date of delivery ≥ 35 years or pre-pregnancy body mass index(BMI)≥24kg/m2were selected.They were randomly divided into intervention group(86 cases)and control group(94 cases).The intervention group,took 1200IU/d vitamin D3 capsules orally from the first trimester of pregnancy to the second trimester of pregnancy[the date of oral glucose tolerance test(OGTT)examination for pregnant women];the control group was not given vitamin D3 capsules,and pregnant women with vitamin D deficiency or insufficient were recommended to take vitamin D3 cap-sules or calcium tablets containing vitamin D.Results The levels of vitamin D in the first and second trimester of pregnancy were 12.986± 5.654ng/ml and 31.277±9.856ng/ml in the intervention group,and the difference was statistically significant(P＜0.001).The inci-dence of gestational diabetes mellitus in control group and intervention group was 24.5％and 20.9％,and the difference was not statisti-cally significant(P＞0.05).There were no significant difference in fasting blood glucose,fasting insulin,1h blood glucose,2h blood glucose and homeostasis model assessment insulin resistance index(HOMR-IR)between the intervention group and the control group in the second trimester of pregnancy(P＞0.05).Conclusion Vitamin D supplementation in early pregnancy can only significantly improve the level of vitamin D in pregnant women,and almost make it reach sufficient state,but has no obvious effect on the risk of gestational dia-betes mellitus and glucose metabolism.

Objective To explore the role of cancer stem cells in EMT-induced acquired resistance to EGFR-TKIs in NSCLC. Methods The EGFR del E746-A750 mutated human lung adenocarcinoma PC-9 cell line and gefitinib acquired resistance cell line PC-9/AB were used in this study. EMT was assessed by western blotting assay. The sensitivity to gefitinib was tested with CCK8. Flow cytometry for antibody analysis was used to quantify CSCs within the cell lines. Results Compared with PC-9, PC-9/AB underwent EMT and showed no-table resistance to gefitinib (P < 0.01). Compared with PC-9, the proportions of CSCs were much higher in PC-9/AB. Conclusion EMT plays an important role in the acquired resistance to EGFR-TKIs in NSCLC , possibly through the up-regulation of CSCs.