Objective:To explore the safety key points of total pancreaticoduodenectomy in the era of vascular resection technology and the important factors affecting rapid postoperative recovery.Methods:The clinical data of 52 patients with pancreatic cancer who underwent total pancreaticoduodenectomy in Beijing Chaoyang Hospital Affiliated to Capital Medical University from November 2014 to September 2022 were retrospectively analyzed, including 34 males and 18 females, aged (62±9). The intraoperative situation, incidence of postoperative complication, postoperative blood glucose control and postoperative survival rate were analyzed.Results:All operations of the 52 patients were successfully completed, including 48 patients underwent total pancreaticoduodenectomy combined with portal vein resection and allograft vascular grafts via artery approach. The portal vein occlusion time was (20±5) min. The incidence of postoperative complications was 28.8% (15/52), including 2 cases of abnormal gastric empty, 2 cases of diarrhea, 2 cases of chylous fistula, 4 cases of abdominal infection, 1 case of gastrointestinal fistula, 3 cases of gastrointestinal bleeding, and 1 case of pulmonary infection. Subcutaneous short-acting insulin injection was used to control blood glucose in the early stage after surgery, short-acting insulin combined with long-acting insulin was used for subcutaneous injection before sleep for diet recovery. All patients did not experience uncontrolled hyperglycemia. The median survival time of 52 patients was 13 months, and the longest follow-up time was 38 months. There were 37 patients died of tumor recurrence, 4 patients died of cardiovascular and cerebrovascular accidents, and 1 patient died of pulmonary infection in the 42 died patients.Conclusions:Total pancreaticoduodenectomy via artery approach can improve the R 0 resection rate in pancreatic cancer patients with vascular invasion, the rate of postoperative complication and mortality has no significant increase. The postoperative blood sugar control is satisfactory and the quality of life is guaranteed.

Objective:To explore the impacts of cyclic RNA asparagine hydroxylase(CircASPH)targeting the miR-28-5p/insulin-like growth factor 1 receptor(IGF-1R)axis on the proliferation,migration,and invasion of ovari-an granulosa cells in polycystic ovary syndrome(PCOS).Methods:Human ovarian granulosa cells KGN and COV434 were used as research subjects,the targeting relationship among CircASPH,miR-28-5p,and IGF-1R was confirmed through dual luciferase reporter gene experiments and pull down experiments.KGN and COV434 cells were grouped into si-NC group,si-ASPH group,si-ASPH+anti NC group,and si-ASPH+anti miR-28-5p group.The mRNA expression levels of CircASPH,miR-28-5p,and IGF-1R mRNA were detected by qRT-PCR,cell proliferation,migration,and invasion were detected by MTT assay,Edu staining,and transwell cell assay,respec-tively;and Western blot was applied to detect the expression of proliferating cell nuclear antigen(PCNA),matrix metalloproteinase-2(MMP-2),vimentin,N-cadherin,E-cadherin,and IGF-1R proteins.Results:Bioinformatics a-nalysis and dual luciferase reporter gene experiments showed that CircASPH,IGF-1R and miR-28-5p had targe-ted binding sites.Compared with si-NC group,the expression level of CircASPH,OD490 value,Edu positive cell rate,cell migration and invasion number,MMP-2,vimentin and N-cadherin in si-ASPH group were decreased,while the expression level of miR-28-5p and E-cadherin protein were increased,and the differences were statisti-cally significant(P<0.05).Compared with the si-ASPH+anti-NC group,the expression level of miR-28-5p and E-cadherin in the si-ASPH+anti-miR-28-5p group were decreased,and the OD490 value,Edu positive cell rate,cell migration and invasion number,MMP-2,vimentin and N-cadherin were increased,the differences were statisti-cally significant(P<0.05).Conclusions:In KGN and COV434 cells,inhibiting the expression of CircASPH can inhibit the proliferation,migration,invasion,and epithelial-mesenchymal transition of ovarian granulosa cells by reg-ulating the miR-28-5p/IGF-1R axis,which may become a new target for the treatment of PCOS.

Objective:To explore the impacts of cyclic RNA asparagine hydroxylase(CircASPH)targeting the miR-28-5p/insulin-like growth factor 1 receptor(IGF-1R)axis on the proliferation,migration,and invasion of ovari-an granulosa cells in polycystic ovary syndrome(PCOS).Methods:Human ovarian granulosa cells KGN and COV434 were used as research subjects,the targeting relationship among CircASPH,miR-28-5p,and IGF-1R was confirmed through dual luciferase reporter gene experiments and pull down experiments.KGN and COV434 cells were grouped into si-NC group,si-ASPH group,si-ASPH+anti NC group,and si-ASPH+anti miR-28-5p group.The mRNA expression levels of CircASPH,miR-28-5p,and IGF-1R mRNA were detected by qRT-PCR,cell proliferation,migration,and invasion were detected by MTT assay,Edu staining,and transwell cell assay,respec-tively;and Western blot was applied to detect the expression of proliferating cell nuclear antigen(PCNA),matrix metalloproteinase-2(MMP-2),vimentin,N-cadherin,E-cadherin,and IGF-1R proteins.Results:Bioinformatics a-nalysis and dual luciferase reporter gene experiments showed that CircASPH,IGF-1R and miR-28-5p had targe-ted binding sites.Compared with si-NC group,the expression level of CircASPH,OD490 value,Edu positive cell rate,cell migration and invasion number,MMP-2,vimentin and N-cadherin in si-ASPH group were decreased,while the expression level of miR-28-5p and E-cadherin protein were increased,and the differences were statisti-cally significant(P<0.05).Compared with the si-ASPH+anti-NC group,the expression level of miR-28-5p and E-cadherin in the si-ASPH+anti-miR-28-5p group were decreased,and the OD490 value,Edu positive cell rate,cell migration and invasion number,MMP-2,vimentin and N-cadherin were increased,the differences were statisti-cally significant(P<0.05).Conclusions:In KGN and COV434 cells,inhibiting the expression of CircASPH can inhibit the proliferation,migration,invasion,and epithelial-mesenchymal transition of ovarian granulosa cells by reg-ulating the miR-28-5p/IGF-1R axis,which may become a new target for the treatment of PCOS.

Objective:To explore the impacts of cyclic RNA asparagine hydroxylase(CircASPH)targeting the miR-28-5p/insulin-like growth factor 1 receptor(IGF-1R)axis on the proliferation,migration,and invasion of ovari-an granulosa cells in polycystic ovary syndrome(PCOS).Methods:Human ovarian granulosa cells KGN and COV434 were used as research subjects,the targeting relationship among CircASPH,miR-28-5p,and IGF-1R was confirmed through dual luciferase reporter gene experiments and pull down experiments.KGN and COV434 cells were grouped into si-NC group,si-ASPH group,si-ASPH+anti NC group,and si-ASPH+anti miR-28-5p group.The mRNA expression levels of CircASPH,miR-28-5p,and IGF-1R mRNA were detected by qRT-PCR,cell proliferation,migration,and invasion were detected by MTT assay,Edu staining,and transwell cell assay,respec-tively;and Western blot was applied to detect the expression of proliferating cell nuclear antigen(PCNA),matrix metalloproteinase-2(MMP-2),vimentin,N-cadherin,E-cadherin,and IGF-1R proteins.Results:Bioinformatics a-nalysis and dual luciferase reporter gene experiments showed that CircASPH,IGF-1R and miR-28-5p had targe-ted binding sites.Compared with si-NC group,the expression level of CircASPH,OD490 value,Edu positive cell rate,cell migration and invasion number,MMP-2,vimentin and N-cadherin in si-ASPH group were decreased,while the expression level of miR-28-5p and E-cadherin protein were increased,and the differences were statisti-cally significant(P<0.05).Compared with the si-ASPH+anti-NC group,the expression level of miR-28-5p and E-cadherin in the si-ASPH+anti-miR-28-5p group were decreased,and the OD490 value,Edu positive cell rate,cell migration and invasion number,MMP-2,vimentin and N-cadherin were increased,the differences were statisti-cally significant(P<0.05).Conclusions:In KGN and COV434 cells,inhibiting the expression of CircASPH can inhibit the proliferation,migration,invasion,and epithelial-mesenchymal transition of ovarian granulosa cells by reg-ulating the miR-28-5p/IGF-1R axis,which may become a new target for the treatment of PCOS.

Objective:To explore the impacts of cyclic RNA asparagine hydroxylase(CircASPH)targeting the miR-28-5p/insulin-like growth factor 1 receptor(IGF-1R)axis on the proliferation,migration,and invasion of ovari-an granulosa cells in polycystic ovary syndrome(PCOS).Methods:Human ovarian granulosa cells KGN and COV434 were used as research subjects,the targeting relationship among CircASPH,miR-28-5p,and IGF-1R was confirmed through dual luciferase reporter gene experiments and pull down experiments.KGN and COV434 cells were grouped into si-NC group,si-ASPH group,si-ASPH+anti NC group,and si-ASPH+anti miR-28-5p group.The mRNA expression levels of CircASPH,miR-28-5p,and IGF-1R mRNA were detected by qRT-PCR,cell proliferation,migration,and invasion were detected by MTT assay,Edu staining,and transwell cell assay,respec-tively;and Western blot was applied to detect the expression of proliferating cell nuclear antigen(PCNA),matrix metalloproteinase-2(MMP-2),vimentin,N-cadherin,E-cadherin,and IGF-1R proteins.Results:Bioinformatics a-nalysis and dual luciferase reporter gene experiments showed that CircASPH,IGF-1R and miR-28-5p had targe-ted binding sites.Compared with si-NC group,the expression level of CircASPH,OD490 value,Edu positive cell rate,cell migration and invasion number,MMP-2,vimentin and N-cadherin in si-ASPH group were decreased,while the expression level of miR-28-5p and E-cadherin protein were increased,and the differences were statisti-cally significant(P<0.05).Compared with the si-ASPH+anti-NC group,the expression level of miR-28-5p and E-cadherin in the si-ASPH+anti-miR-28-5p group were decreased,and the OD490 value,Edu positive cell rate,cell migration and invasion number,MMP-2,vimentin and N-cadherin were increased,the differences were statisti-cally significant(P<0.05).Conclusions:In KGN and COV434 cells,inhibiting the expression of CircASPH can inhibit the proliferation,migration,invasion,and epithelial-mesenchymal transition of ovarian granulosa cells by reg-ulating the miR-28-5p/IGF-1R axis,which may become a new target for the treatment of PCOS.

Objective:To explore the impacts of cyclic RNA asparagine hydroxylase(CircASPH)targeting the miR-28-5p/insulin-like growth factor 1 receptor(IGF-1R)axis on the proliferation,migration,and invasion of ovari-an granulosa cells in polycystic ovary syndrome(PCOS).Methods:Human ovarian granulosa cells KGN and COV434 were used as research subjects,the targeting relationship among CircASPH,miR-28-5p,and IGF-1R was confirmed through dual luciferase reporter gene experiments and pull down experiments.KGN and COV434 cells were grouped into si-NC group,si-ASPH group,si-ASPH+anti NC group,and si-ASPH+anti miR-28-5p group.The mRNA expression levels of CircASPH,miR-28-5p,and IGF-1R mRNA were detected by qRT-PCR,cell proliferation,migration,and invasion were detected by MTT assay,Edu staining,and transwell cell assay,respec-tively;and Western blot was applied to detect the expression of proliferating cell nuclear antigen(PCNA),matrix metalloproteinase-2(MMP-2),vimentin,N-cadherin,E-cadherin,and IGF-1R proteins.Results:Bioinformatics a-nalysis and dual luciferase reporter gene experiments showed that CircASPH,IGF-1R and miR-28-5p had targe-ted binding sites.Compared with si-NC group,the expression level of CircASPH,OD490 value,Edu positive cell rate,cell migration and invasion number,MMP-2,vimentin and N-cadherin in si-ASPH group were decreased,while the expression level of miR-28-5p and E-cadherin protein were increased,and the differences were statisti-cally significant(P<0.05).Compared with the si-ASPH+anti-NC group,the expression level of miR-28-5p and E-cadherin in the si-ASPH+anti-miR-28-5p group were decreased,and the OD490 value,Edu positive cell rate,cell migration and invasion number,MMP-2,vimentin and N-cadherin were increased,the differences were statisti-cally significant(P<0.05).Conclusions:In KGN and COV434 cells,inhibiting the expression of CircASPH can inhibit the proliferation,migration,invasion,and epithelial-mesenchymal transition of ovarian granulosa cells by reg-ulating the miR-28-5p/IGF-1R axis,which may become a new target for the treatment of PCOS.

Objective:To explore the impacts of cyclic RNA asparagine hydroxylase(CircASPH)targeting the miR-28-5p/insulin-like growth factor 1 receptor(IGF-1R)axis on the proliferation,migration,and invasion of ovari-an granulosa cells in polycystic ovary syndrome(PCOS).Methods:Human ovarian granulosa cells KGN and COV434 were used as research subjects,the targeting relationship among CircASPH,miR-28-5p,and IGF-1R was confirmed through dual luciferase reporter gene experiments and pull down experiments.KGN and COV434 cells were grouped into si-NC group,si-ASPH group,si-ASPH+anti NC group,and si-ASPH+anti miR-28-5p group.The mRNA expression levels of CircASPH,miR-28-5p,and IGF-1R mRNA were detected by qRT-PCR,cell proliferation,migration,and invasion were detected by MTT assay,Edu staining,and transwell cell assay,respec-tively;and Western blot was applied to detect the expression of proliferating cell nuclear antigen(PCNA),matrix metalloproteinase-2(MMP-2),vimentin,N-cadherin,E-cadherin,and IGF-1R proteins.Results:Bioinformatics a-nalysis and dual luciferase reporter gene experiments showed that CircASPH,IGF-1R and miR-28-5p had targe-ted binding sites.Compared with si-NC group,the expression level of CircASPH,OD490 value,Edu positive cell rate,cell migration and invasion number,MMP-2,vimentin and N-cadherin in si-ASPH group were decreased,while the expression level of miR-28-5p and E-cadherin protein were increased,and the differences were statisti-cally significant(P<0.05).Compared with the si-ASPH+anti-NC group,the expression level of miR-28-5p and E-cadherin in the si-ASPH+anti-miR-28-5p group were decreased,and the OD490 value,Edu positive cell rate,cell migration and invasion number,MMP-2,vimentin and N-cadherin were increased,the differences were statisti-cally significant(P<0.05).Conclusions:In KGN and COV434 cells,inhibiting the expression of CircASPH can inhibit the proliferation,migration,invasion,and epithelial-mesenchymal transition of ovarian granulosa cells by reg-ulating the miR-28-5p/IGF-1R axis,which may become a new target for the treatment of PCOS.

Objective:To explore the impacts of cyclic RNA asparagine hydroxylase(CircASPH)targeting the miR-28-5p/insulin-like growth factor 1 receptor(IGF-1R)axis on the proliferation,migration,and invasion of ovari-an granulosa cells in polycystic ovary syndrome(PCOS).Methods:Human ovarian granulosa cells KGN and COV434 were used as research subjects,the targeting relationship among CircASPH,miR-28-5p,and IGF-1R was confirmed through dual luciferase reporter gene experiments and pull down experiments.KGN and COV434 cells were grouped into si-NC group,si-ASPH group,si-ASPH+anti NC group,and si-ASPH+anti miR-28-5p group.The mRNA expression levels of CircASPH,miR-28-5p,and IGF-1R mRNA were detected by qRT-PCR,cell proliferation,migration,and invasion were detected by MTT assay,Edu staining,and transwell cell assay,respec-tively;and Western blot was applied to detect the expression of proliferating cell nuclear antigen(PCNA),matrix metalloproteinase-2(MMP-2),vimentin,N-cadherin,E-cadherin,and IGF-1R proteins.Results:Bioinformatics a-nalysis and dual luciferase reporter gene experiments showed that CircASPH,IGF-1R and miR-28-5p had targe-ted binding sites.Compared with si-NC group,the expression level of CircASPH,OD490 value,Edu positive cell rate,cell migration and invasion number,MMP-2,vimentin and N-cadherin in si-ASPH group were decreased,while the expression level of miR-28-5p and E-cadherin protein were increased,and the differences were statisti-cally significant(P<0.05).Compared with the si-ASPH+anti-NC group,the expression level of miR-28-5p and E-cadherin in the si-ASPH+anti-miR-28-5p group were decreased,and the OD490 value,Edu positive cell rate,cell migration and invasion number,MMP-2,vimentin and N-cadherin were increased,the differences were statisti-cally significant(P<0.05).Conclusions:In KGN and COV434 cells,inhibiting the expression of CircASPH can inhibit the proliferation,migration,invasion,and epithelial-mesenchymal transition of ovarian granulosa cells by reg-ulating the miR-28-5p/IGF-1R axis,which may become a new target for the treatment of PCOS.

Objective:To explore the impacts of cyclic RNA asparagine hydroxylase(CircASPH)targeting the miR-28-5p/insulin-like growth factor 1 receptor(IGF-1R)axis on the proliferation,migration,and invasion of ovari-an granulosa cells in polycystic ovary syndrome(PCOS).Methods:Human ovarian granulosa cells KGN and COV434 were used as research subjects,the targeting relationship among CircASPH,miR-28-5p,and IGF-1R was confirmed through dual luciferase reporter gene experiments and pull down experiments.KGN and COV434 cells were grouped into si-NC group,si-ASPH group,si-ASPH+anti NC group,and si-ASPH+anti miR-28-5p group.The mRNA expression levels of CircASPH,miR-28-5p,and IGF-1R mRNA were detected by qRT-PCR,cell proliferation,migration,and invasion were detected by MTT assay,Edu staining,and transwell cell assay,respec-tively;and Western blot was applied to detect the expression of proliferating cell nuclear antigen(PCNA),matrix metalloproteinase-2(MMP-2),vimentin,N-cadherin,E-cadherin,and IGF-1R proteins.Results:Bioinformatics a-nalysis and dual luciferase reporter gene experiments showed that CircASPH,IGF-1R and miR-28-5p had targe-ted binding sites.Compared with si-NC group,the expression level of CircASPH,OD490 value,Edu positive cell rate,cell migration and invasion number,MMP-2,vimentin and N-cadherin in si-ASPH group were decreased,while the expression level of miR-28-5p and E-cadherin protein were increased,and the differences were statisti-cally significant(P<0.05).Compared with the si-ASPH+anti-NC group,the expression level of miR-28-5p and E-cadherin in the si-ASPH+anti-miR-28-5p group were decreased,and the OD490 value,Edu positive cell rate,cell migration and invasion number,MMP-2,vimentin and N-cadherin were increased,the differences were statisti-cally significant(P<0.05).Conclusions:In KGN and COV434 cells,inhibiting the expression of CircASPH can inhibit the proliferation,migration,invasion,and epithelial-mesenchymal transition of ovarian granulosa cells by reg-ulating the miR-28-5p/IGF-1R axis,which may become a new target for the treatment of PCOS.

Objective·To establish a cell-based screening system for identification of compounds with activity in regulating retinoic acid receptor (RARα) stability. Methods·The modified pMSCV plasmid constructs, named as RARα-EGFP-IRES-DsRed, consists of enhanced green fluorescent protein (EGFP) fusing to RARα and red fluorescent protein (DsRed) as internal references incorporating the internal ribosome entry site (IRES) as interval sequence. The RARα-EGFP-IRES-DsRed plasmid was stably transfected into NB4 cells which were named as NB4-pMGIR-RARα. Fluorescence signals of EGFP and DsRed indirectly reflecting the expression of RARα, were detected by flow cytometry in cells that were treated with all-trans retinoic acid, sodium valproate, cytarabine, lenalidomide, etoposide, montelukast and gambogic acid, respectively. Effects of these compounds on the expression of RARα protein were further examined by Western blotting. Results·A double fluorescence reporter system for screening compounds that can increase the stability of RARα protein was successfully established, and sodium valproate was identified as a potent compound to promote the stability of RARα. Conclusion·The double fluorescence reporter system can be used to screen compounds regulating the stability of RARα protein, which can be further used to identify compounds regulating the stability of other proteins.