Receptor-interacting protein kinase 1 (RIPK1) plays an essential role in regulating the necroptosis and apoptosis in cerebral ischemia-reperfusion (I/R) injury. However, the regulation of RIPK1 kinase activity after cerebral I/R injury remains largely unknown. In this study, we found the downregulation of protein arginine methyltransferase 1 (PRMT1) was induced by cerebral I/R injury, which negatively correlated with the activation of RIPK1. Mechanistically, we proved that PRMT1 directly interacted with RIPK1 and catalyzed its asymmetric dimethylarginine, which then blocked RIPK1 homodimerization and suppressed its kinase activity. Moreover, pharmacological inhibition or genetic ablation of PRMT1 aggravated I/R injury by promoting RIPK1-mediated necroptosis and apoptosis, while PRMT1 overexpression protected against I/R injury by suppressing RIPK1 activation. Our findings revealed the molecular regulation of RIPK1 activation and demonstrated PRMT1 would be a potential therapeutic target for the treatment of ischemic stroke.

Objective:To explore the characteristics of echocardiographic parameters among the many parameters of two-dimensional transthoracic echocardiography(2D-TTE) and three-dimensional speckle tracking imaging (3D-STI) that can be used for early identification of familial hypertrophic cardiomyopathy(FHCM) mutation gene carriers, and construct a Nomogram prediction model, in order to provide a diagnostic method for early identification of G+ P- patients for clinical practice.Methods:A total of 15 FHCM families admitted to the General Hospital of Ningxia Medical University from November 2017 to August 2022 were enrolled.Whole exome sequencing and Sanger sequencing technology were used for gene detection, among which 54 were G+ P- and 75 were G-P-. Stratified random sampling was used to divide the subjects into training set ( n=90) and test set ( n=39) according to the ratio of 7∶3. Philips iE33 ultrasonic diagnostic instrument and TomTec offline software were used to obtain relevant ultrasonic parameters. Lasso regression and Logistic regression were used to screen echocardiographic parameters and obtain independent risk factors for early prediction of G+ P-, based on which a Nomogram prediction model was established. Results:①Lasso-Logistic regression showed that global longitudinal strain(GLS) ( OR=1.739, 95% CI=1.305-2.316) and left ventricular outflow trac velocity time integral(LVOT-VTI) ( OR=1.358, 95% CI=1.072-1.722) could be used as independent risk factors for early prediction of G+ P-. ②The Nomogram prediction model was established based on the above indicators. After 1000 internal verifications of Bootstrap self-sampling, the C-indices of the training set and the test set were 0.885 (95% CI=0.816-0.954), 0.878 (95% CI=0.764-0.992), which had good internal consistency. ③The results of the calibration curve showed that the risk of G+ P- predicted by the Nomogram model was basically consistent with the actual risk (training set P=0.990, test set P=0.961); the clinical decision curve shows that under different threshold probabilities, using this prediction model to provide patients with clinical decision-making could bring benefits to patients. Conclusions:Echocardiographic parameters GLS and LVOT-VTI can be used as independent risk factors to predict FHCM mutation gene carriers. The Nomogram prediction model has good discrimination, goodness of fit and clinical benefit in identifying whether the family members of FHCM patients carry the mutation gene, and it can provide a new idea and evaluation method for the early identification of FHCM mutation gene carriers by echocardiography.

Background: Currently available guidelines contain conflicting recommendations on the management of blood pressure (BP) in patients with diabetes mellitus (DM). Therefore, it is necessary to appraise the guidelines and summarize the agreements and differences among recommendations. Methods: Four databases and the websites of guideline organizations were searched for guidelines regarding BP targets and thresholds for pharmacologic therapy in DM patients, and the included guidelines were appraised with the Appraisal of Guidelines for Research and Evaluation (AGREE) II instrument. Results: In 6,498 records identified, 20 guidelines met our inclusion criteria with 64.0% AGREE II scores (interquartile range, 48.5% to 72.0%). The scores of the European and American guidelines were superior to those of the Asian guidelines (both adjusted P<0.001). Most of the guidelines advocated systolic BP targets <130 mm Hg (12 guidelines, 60%) and diastolic BP targets <80 mm Hg (14 guidelines, 70%) in DM patients. Approximately half of the guidelines supported systolic BP thresholds >140 mm Hg (10 guidelines, 50%) and diastolic BP thresholds >90 mm Hg (nine guidelines, 45%). The tiny minority of the guidelines provided the relevant recommendations regarding the lower limit of official BP targets and the ambulatory BP monitoring (ABPM)/home BP monitoring (HBPM) targets and thresholds in DM patients. Conclusion The lower official BP targets (<130/80 mm Hg) in patients with DM are advocated by most of the guidelines, but they contain conflicting recommendations on the official BP thresholds. Moreover, the gaps regarding the lower limit of official BP targets and the ABPM/HBPM targets and thresholds need to be considered by future study.

Background: Currently available guidelines contain conflicting recommendations on the management of blood pressure (BP) in patients with diabetes mellitus (DM). Therefore, it is necessary to appraise the guidelines and summarize the agreements and differences among recommendations. Methods: Four databases and the websites of guideline organizations were searched for guidelines regarding BP targets and thresholds for pharmacologic therapy in DM patients, and the included guidelines were appraised with the Appraisal of Guidelines for Research and Evaluation (AGREE) II instrument. Results: In 6,498 records identified, 20 guidelines met our inclusion criteria with 64.0% AGREE II scores (interquartile range, 48.5% to 72.0%). The scores of the European and American guidelines were superior to those of the Asian guidelines (both adjusted P<0.001). Most of the guidelines advocated systolic BP targets <130 mm Hg (12 guidelines, 60%) and diastolic BP targets <80 mm Hg (14 guidelines, 70%) in DM patients. Approximately half of the guidelines supported systolic BP thresholds >140 mm Hg (10 guidelines, 50%) and diastolic BP thresholds >90 mm Hg (nine guidelines, 45%). The tiny minority of the guidelines provided the relevant recommendations regarding the lower limit of official BP targets and the ambulatory BP monitoring (ABPM)/home BP monitoring (HBPM) targets and thresholds in DM patients. Conclusion The lower official BP targets (<130/80 mm Hg) in patients with DM are advocated by most of the guidelines, but they contain conflicting recommendations on the official BP thresholds. Moreover, the gaps regarding the lower limit of official BP targets and the ABPM/HBPM targets and thresholds need to be considered by future study.

Background:Abnormal expression of long non-coding RNAs(lncRNAs)has been found in almost all tumors in humans,providing numerous potential diagnostic and prognostic biomarkers,and therapeutic targets. Materials and methods:The Cancer Genome Atlas(TCGA)database was used to screen potential LncRNAs,and 30 paired hepatocellular carcinoma(HCC)tissues were used to investigate RP11-307C12.11 expression levels by qRT-PCR and another 105 HCC tissues by in situ hybridizsation(ISH).RP11-307C12.11 overexpression and knockdown experiments were performed to investigate the effects of RP11-307C12.11 on HCC growth through in vitro and in vivo assays(MTT assay,colony formation assay,EdU assay,and xenograft model).The molecular mechanism underlying these effects was confirmed by MS2-RIP-assay,RIP assay,luciferase assay,and rescue experiments. Results:RP11-307C12.11 expression level was significantly higher in tumor tissues than in the adjacent normal tissues.Elevated RP11-307C12.11 expression level was associated with poor prognosis of HCC patients,and it may be represented as an independent prognostic biomarker in patients with HCC.Functionally,RP11-307C12.11 overexpression promoted HCC growth both in vitro and in vivo;however,its knockdown reversed these effects.Mechanistically,we found that RP11-307C12.11 expressed pre-dominantly in the cytoplasm and sponged microRNA(miR)-138 to regulate its common target CCND1 and PDK1. Conclusions:Thus,we found that RP11-307C12.11 acts as an oncogene in HCC by binding to miR-138,which might provide a novel target for HCC therapy.

Objective To compare the clinical outcomes and complications of alloplasfic cranioplasty performed with custom-designed polyetheretherketone (PEEK) and titanium mesh after decompressive craniectomy.Methods Eighty-six patients admitted to our hospital from June 2014 to December 2017 were chosen;and 28 patients underwent cranioplasty with PEEK and 58 with titanium mesh by the same surgical team.The general clinical data and postoperative complications were compared between the two groups.Multivariable Logistic regression analysis was performed to analyze the influencing factors of postoperative complications.The surgical time,molding quality and cost were compared between the two groups.Results Patients in PEEK group trended to be younger and had higher GOS scores as compared with patients in the titanium group,with significant differences (P＜0.05).Overall complication rates of 10.7％ and 32.8％ for PEEK and titanium cranioplasty were identified respectively;as compared with that in titanium group,the incidence of overall complication in PEEK group was significant lower (P＜0.05).Logistic regression analysis identified material was the independent influencing factor for cranioplasty complications (OR=4.486,P=0.047,95％CI:1.021-19.703).Overall satisfaction rate with cranioplasty and aesthetic result in PEEK group was significantly higher than that in titanium group (96.4％ vs.79.3％,P＜0.05);however,the treatment cost for cranioplasty with PEEK was considerably higher than skull bone reconstruction based on titanium mesh.Conclusion Despite of high treatment cost,custom-designed PEEK implants seem to be good choice for patients with large cranial defects after decompressive craniectomy,enjoying few complications and high satisfaction of cranioplasty and aesthetic result.

Objective To explore the expression,role and mechanism of long non-coding RNA (lncRNA) BANCR in astrocytoma.Methods (1) Twenty-four astrocytoma tissues and 6 normal brain tissues were collected from patients accepted surgical resection and conformed by pathology in our hospital from January 2016 to September 2017;the mRNA expressions of BANCR and signal transduction and transcriptional activator 3 (STA T3) were detected by real-time quantitative(qRT)-PCR;the glioma dataset GSE4290 including astrocytomas were downloaded and BANCR expressions in GSE4290 were analyzed by the web software R2.(2) Astrocytoma cell line LN18,cultured in vitro were divided into short hairpin RNA (shBANCR) group,full-length BANCR (BANCR) group,shControl group and empty vector group;cells in these groups were transfected with recombinant lentiviruses packing genomes encoding short hairpin RNA (shBANCR),full-length BANCR (BANCR),or their corresponding controls (shControl and empty vector);BANCR and STA T3 mRNA expressions in the 4 groups were detected by qRT-PCR;the cell proliferation,migration and invasion,and apoptosis were detected by CCK-8 assay,Transwell assay and flow cytometry,respectively;Western blotting was employed to detect the protein expressions of STAT3,matrix metalloproteinase (MMP)2,MMP9 and mitogen-activated protein kinase (MAPK),and Akt pathway protein expression.(3) Astrocytoma cell line LN18 were divided into si398 group,si1265 group,negative control group Ⅰ,and blank control group Ⅰ;cells in the blank control group Ⅰ were transfected with lipofectamine 2000,and cells in the other three groups were transfected with small interfering RNA si398,si1265 and negative control nonsense sequences targeting STAT3;48 h after transfection,BANCR and STAT3 mRNA expressions were detected by qRT-PCR;Western blotting was employed to detect the STAT3 protein expression.Results (1) In collected samples and glioma dataset GSE4290:the BANCR mRNA expression in astrocytoma tissues was significantly decreased as compared with that in the normal brain tissues (P＜0.05);a positive correlation was noted between BANCR and STA T3 mRNA expressions in astrocytomas (P＜0.05).(2) As compared with the negative control group,the shBANCR group had significantly decreased BANCR mRNA expression,and the BANCR mRNA expression in the BANCR group was significantly increased as compared with that in the empty vector group (P＜0.05);the number of migration and invasion cells in the shBANCR group was significantly larger as compared with that in the negative control group,and that in the BANCR group was significantly increased as compared with that in the empty vector group (P＜0.05);the protein levels of MMP2,MMP9,phosphorylated (p)-extracellular regulated protein kinase and p-mitogen-activated protein kinase in the shBANCR group were significantly increased as compared with those in the negative control group,and those in the BANCR group was significantly increased as compared with those in the empty vector group (P＜0.05).(3) As compared with negative control group Ⅰ and blank control group Ⅰ,si398 group and si1265 group had significantly decreased STA T3 and BANCR mRNA expressions and STAT3 protein expression (P＜0.05).Conclusion The BANCR expression is decreased in astrocytoma;STAT3-induced BANCR can inhibit cell migration and invasion by modulating MMP2,MMP9 and MAPK signaling pathway in astrocytoma.

Objective To explore the effect of fibroblast growth factor receptor(FGFR)inhibitor BGJ398 on the vasculogenic mimicry(VM)formation of glioma cells. Methods The phosphor-FGFR(pFGFR)was de-tected by Western blot,the expressions of MMP2 and MMP14 were detected by Western blot and immunocytochem-istry;the VM formation of U87MG and U251MG was tested by tube formation assay;subcutaneously implanted tu-mor model in nude mouse was established and tumor sections were CD34/PAS double-stained to detect the forma-tion of VM in vivo. Results Western blot showed that pFGFR in the experimental groups decreased significantly (P < 0.05);western blot and immunohistochemical staining showed that the expression of MMP2 and MMP14 in the experimental groups decreased significantly compared to the control group. In the tube formation assay ,the tube formation of U87MG and U251MG cells were restrained. In the subcutaneously implanted tumor model ,the VM number of the experimental group(13.85 ± 3.96)was significantly lower than that in the control group(26.40 ± 5.06,P < 0.05). Conclusions In vivo and in vitro experiments confirmed that BGJ398 can inhibit the activa-tion of FGFR,and inhibit the VM formation of glioma cells. These indicate FGFR signaling pathway is involved in the formation of VM.

Objective To study the transcriptional regulation of vp1667 by H-NS in Vibrio parahaemolyticus.Methods Total RNAs were extracted from Δhns and WT strains.Quantitative RT-PCR was carried out to calculate the transcriptional variation of vp1667 between Δhns and WT.Primer extension assay was also employed to detect the transcription start site and the promoter activity (i.e.the amount of primer extension products) of vp1667 in Δhns and that in WT.The promoter DNA region of vp1667 was amplified, purified, and cloned into the corresponding restriction endonuclease sites of pHRP309 that harbors a gentamicin resistance gene and a promoterless lacZ reporter gene.The recombinant pHRP309 plasmid was transformed into Δhns and WT, respectively, while β-galactosidase activity in cellular extracts was measured using a β-galactosidase enzyme assay system.The over-expressed His-H-NS was purified under native conditions with nickel loaded HiTrap Chelating Sepharose columns.The electrophoretic mobility shift assay (EMSA) and DNaseⅠ footprinting were then applied to analyze the DNA-binding activity of His-H-NS to vp1667 promoter region in vitro.Results and Conclusion The primer extension assay detected one transcription start site for vp1667, which was located at 28 bp upstream of vp1667, and its transcribed activity was under the negative control of the H-NS.The EMSA and DNaseⅠ footprinting assay results showed that His-H-NS was unable to bind to the promoter-proximal DNA region of vp1667, suggesting that H-NS indirectly inhibits the transcription of vp1667.

Objective To investigate the transcriptional autoregulation of PhoP/PhoQ under different growth conditions in Yersinia pestis.Methods The entire promoter region of YPO1635 was amplified and cloned into the pRW50 vector containing a promoterless lacZ reporter gene.The recombinant LacZ reporter plasmid was transformed into the wild-type strain (WT) and the phoP mutant strain (ΔphoP),respectively,to measure the promoter activity (the β-galactosidase activity) of the target gene in WT and ΔphoP by using the β-galactosidase enzyme assay system.Total RNAs were extracted from WT and ΔphoP strains,and primer extension assay was employed to detect the promoter activity by examining the amount of primer extension products of YPO1635 in WT and ΔphoP.Results The LacZ fusion results showed that the transcription of YPO1635 was positively regulated by PhoP under L-TMH and brain-heart infusion(BHI) conditions,but it was not regulated in H-TMH medium.The primer extension assay detected two transcriptional start sites located at 90 and 118 bp upstream of the translation initiation site of phoP,named P1 and P2,respectively.Under low Mg2+ TMH conditions,the promoter activity of P1 rather than P2 was positively regulated by PhoP.Under high Mg2+ TMH conditions,the promoter activities of both P1 and P2 showed no obvious difference in the WT and ΔphoP strains.Under rich BHI conditions,both promoters were under negative control of PhoP.Conclusion Different autoregualtion patterns of PhoP/PhoQ under different growth conditions would help Y.pestis to quickly adapt to the changing living environment.