Objective: To explore the effect of YTH domain family protein 1(YTHDF1) on the activation, proliferation and migration of hepatic stellate cells(HSCs) by regulating mitochondrial fission mediated by mitochondrial fission protein 1(Fis1). Methods: The mouse hepatic stellate cell line JS-1 was treated with 5 ng/ml TGF-β1 for 24 h to induce its activation and proliferation, andYTHDF1-siRNA was used to construct aYTHDF1silencing model.The experiment was divided into Control group, TGF-β1 group, TGF-β1+si-NC group and TGF-β1+si-YTHDF1 group.Expression changes ofYTHDF1,Fis1and key indicators of fibrosis, type Ⅰ collagen(CollagenⅠ) and α-smooth muscle actin(α-SMA) were detected through reverse transcription quantitative polymerase chain reaction(RT-qPCR) and Western blot; CCK-8 was used to detect cell proliferation ability; Transwell migration assay and cell scratch assay were used to detect cell migration ability; immunofluorescence staining experiment was used to detect the effect ofYTHDF1onFis1-mediated mitochondrial fission; finally, JC-1 staining was used to experimentally detect the effect ofYTHDF1on mitochondrial membrane potential. Results: Compared with the Control group, RT-qPCR and Western blot experimental results showed that the expression ofYTHDF1andFis1increased in the TGF-β1 group(P<0.05,P<0.01;P<0.000 1), as well as the fibrosis markersCollagenⅠand the expression level of α-SMA increased(P<0.01;P<0.001,P<0.000 1); while adding CCK-8, the experimental results showed that the proliferation ability of HSCs in the TGF-β1 group was enhanced(P<0.000 1); Transwell experimental results showed that the migration ability of HSCs in the TGF-β1 group was enhanced(P<0.01); the cell scratch experiment results showed that the migration ability of HSCs in the TGF-β1 group was enhanced(P<0.000 1); the immunofluorescence experiment results showed that the TGF-β1 group Mito-Tracker Red staining andFis1co-localization signal increased(P<0.05); JC-1 staining experiment results showed that the mitochondrial membrane potential increased in the TGF-β1 group(P<0.01). Compared with the TGF-β1+si-NC group, RT-qPCR and Western blot experimental results showed that the expression ofYTHDF1andFis1in the TGF-β1+si-YTHDF1 group was reduced(P<0.01;P<0.001), and fibrosis markers the levels ofCollagenⅠandα-SMAwere reduced(P<0.01;P<0.001,P<0.01).CCK-8 experimental results showed that the proliferation ability of HSCs in the TGF-β1+si-YTHDF1 group was weakened(P<0.000 1); Transwell experiment results showed that the migration ability of HSCs in the TGF-β1+si-YTHDF1 group was weakened(P<0.001); cell scratch experiment results showed that the migration ability of HSCs in the TGF-β1+si-YTHDF1 group was weakened(P<0.000 1); immunofluorescence experiment results showed that the Mito-Tracker Red staining andFis1co-localization signal decreased in the TGF-β1+si-YTHDF1 group(P<0.01); JC-1 staining experiment results showed that mitochondrial membrane potential decreased in the TGF-β1+si-YTHDF1 group(P<0.05). Conclusion YTHDF1promotes the activation, proliferation and migration capabilities of HSCs by positively regulatingFis1-mediated mitochondrial fission. This suggests thatYTHDF1may be a key gene involved in regulating the activation, proliferation and migration of HSCs.

AIM: To investigate the effect of the timing of satisfactory sedation with preoperative oral midazolam on anesthesia induction and recovery in children undergoing adenotonsillectomy. METHODS: A total of 147 children undergoing elective adenotonsillectomy, with ASA physical status orⅡ, aged 2-7 years were selected from November 2022 to June 2023 in the Second Affiliated Hospital of Wenzhou Medical University. The children were orally administered 0.5 mg/kg midazolam in preoperative waiting area and were divided into 10-20 min (rapid onset, M1 group) and 21-30 min (slow onset, M2 group) based on the satisfactory sedation time, or equal volume of sugar pear drink orally (blank control group, C group). Children in all three groups received a general anesthesia method of propofol+fentanyl combined with sevoflurane induction and sevoflurane maintenance. The primary outcome measures were the induction compliance checklist (ICC) score and the pediatric anesthesia emergence delirium (PAED) score in the post-anesthesia care unit (PACU) to assess the occurrence of emergence agitation (EA), and the secondary outcome measures included the parental separation anxiety scale (PSAS), sedation Ramsay score, surgery duration, recovery time, PACU stay time, discharge time, the incidence of perioperative respiratory adverse events (PRAE) and other adverse events in the ward. RESULTS: 147 children were included in the result analysis, with 49 cases in each group. The proportion of perfect induction (ICC=0) were significantly higher in two M groups than that in group C (95.9% vs. 91.8% vs. 61.2%, P=0.001). The maximum and average PAED score in PACU in group M1 showed a significantly higher (6.4±5.0 vs. 4.4 ± 4.1, P=0.029; 5.2 ± 4.5 vs. 3.4 ± 3.6, P=0.030), and the incidence of EA was significantly higher than those in group C (10.2% vs. 30.6%, P=0.022), and increased compared to the group M2 (OR= 0.581, 95%CI 0.231-1.463, P=0.354). There was no statistically significant difference in the maximum and average PAED scores, incidence of EA between group M2 and group C (P>0.05). The Ramsay score and PSAS score in two M groups were higher, PACU stay time and recovery time was longer than those in group C (P<0.05). The pain scores in PACU in group M1 was higher than that of group C (P<0.05). There was no statistically significant difference in the surgical time, discharge time, the incidence of PRAE and other adverse events in the ward among three groups (P>0.05). CONCLUSION: Preoperative oral midazolam can improve the ICC and PSAS scores of children during induction, but it also leads to prolonged recovery time and PACU retention time. The rapid onset of midazolam did not result in better induction and recovery quality, but instead increased the incidence of EA and postoperative pain score.

Narciclasine(NCS),a hymenocallis littoralis alkaloid extracted from the bulbs of the genus Narcissus in the Lycoriaceae family,has been proven to have significant anti-tumor activity against a variety of tumor cells.The antitumor mechanisms of NCS are diverse and NCS exhibits antitumor effects through different pathways,which adapts to the current trend of developing multi-target anti-tumor drugs.This review introduces the research progress of the anti-tumor activity and mechanism of NCS in recent years based on the inhibitory effect of NCS on gastric cancer cells,oral cancer cells,polymorphous glioblastoma cells,colon cancer cells,breast cancer cells,melanoma cells and primary exudative lymphoma cells,aiming to provide ideas and references for the research and development,and design of NCS type anti-tumor drugs in the future.

BACKGROUND:M2 macrophages have the function of reducing inflammatory factors and promoting tissue healing.Therefore,how to regulate M2 polarization of macrophages has been a hot research topic in recent years,and some miRNAs have been found to have this function. OBJECTIVE:To investigate the effects of miR-378a on the polarization of the Raw264.7 macrophage cell line. METHODS:The M1 polarization of macrophages was induced by lipopolysaccharide and interferon-γ.Interleukin-4 induced M2 polarization and the expression of endogenous miR-378a in each cell type was detected using qRT-PCR to verify whether miR-378a was involved in the polarization of macrophages.By transfection with lentivirus as the vector of overexpression of miR-378a,the stable expression of miR-378a cell lines was screened.Macrophage M1 polarization was induced synergically by lipopolysaccharide and interferon-γ.Macrophage M2 polarization was induced by interleukin-4.The levels of M1/M2 polarization-related cytokines in the supernatant of the macrophage culture medium were determined by enzyme-linked immunosorbent assay.qRT-PCR was used to detect the polarization characteristics of M1/M2-type macrophages and the mRNA expression levels of related cytokines. RESULTS AND CONCLUSION:(1)The expression level of endogenous miR-378a in Raw264.7 cells of each group increased after macrophage polarization.(2)Compared with the non-transfected group,the expressions of proinflammatory cytokine-induced nitric oxide synthase,tumor necrosis factor-α,interleukin-6 and interleukin-1β in macrophage M1 induced polarization were significantly decreased in the miR-378a transfection group(P<0.05);the levels of inducible nitric oxide synthase,tumor necrosis factor-α and interleukin-6 in cell supernatant were also significantly decreased(P<0.05).(3)Compared with the non-transfected group,the expressions of CD206,interleukin-10 and arginase-I in macrophage M2 induced polarization were significantly increased(P<0.05);the levels of CD206 and interleukin-10 in cell supernatant were also significantly increased(P<0.05)in the miR-378a transfection group.(4)It is indicated that overexpression of miR-378a promotes the M2 polarization of macrophages and inhibits the M1 polarization of macrophages.

Objective:To identify the risk factors for 1-year death after surgery in elderly patients with hip fractures and evaluate the accuracy of the prediction model based on LASSO-logistic regression analysis.Methods:A case-control study was conducted on elderly patients (age ≥65 yr) who underwent surgical treatment for hip fractures in the Second Affiliated Hospital of Wenzhou Medical University from January to December 2019. Patients were divided into death group and survival group according to their survival status at 1-year after surgery. General data and preoperative laboratory indicators were obtained. The variables were selected by utilizing LASSO regression and incorporated into multivariate logistic regression analysis to identify the risk factors for 1-year death after surgery in elderly patients with hip fractures. Then a prediction model was established based on the results and evaluated.Results:There were 63 patients in death group and 564 in survival group. The results of LASSO regression and multivariate logistic regression analysis showed that age, preoperative cognitive dysfunction, Chalson comorbidity index ≥3 points and preoperative serum prealbumin level were the independent risk factors for 1-year death after surgery in elderly patients with hip fractures ( P<0.05). The area under the receiver operating characteristic curve of the prediction model was 0.788 (95% confidence interval [0.731-0.846]), with the sensitivity and specificity of 76.2% and 68.6% respectively. The average absolute error of the calibration curve was 0.007. The results of Hosmer-Lemeshow goodness-of-fit test showed that there was no significant difference between the predicted value and actual observed value ( χ2=5.065, P=0.751). Decision curve analysis showed that patients had a high net benefit rate when the threshold probability range was 0-0.7. Conclusions:Age, preoperative cognitive dysfunction, Chalson comorbidity index ≥3 points and preoperative serum prealbumin level are the independent risk factors for 1-year death after surgery in elderly patients with hip fractures, and the prediction model developed based on LASSO-logistic regression has high accuracy.

Objective This study aimed to investigate the effects of sitagliptin on the proliferation,apoptosis,in-flammation,and osteogenic differentiation of human periodontal ligament stem cells(hPDLSCs)in lipopolysaccharide(LPS)-induced inflammatory microenvironment and its molecular mechanism.Methods hPDLSCs were cultured in vitro and treated with different concentrations of sitagliptin to detect cell viability and subsequently determine the exper-imental concentration of sitagliptin.An hPDLSCs inflammation model was established after 24 h of stimulation with 1 μg/mL LPS and divided into blank,control,low-concentration sitagliptin(0.5 μmol/L),medium-concentration sita-gliptin(1 μmol/L),and high-concentration sitagliptin(2 μmol/L),high-concentrationsitagliptin+stromal cell derived factor-1(SDF-1)/CXC chemokine receptor 4(CXCR4)pathway inhibitor(AMD3100)(2 μmol/L+10 μg/mL)groups.A cell-counting kit-8 was used to detect the proliferation activity of hPDLSCs after 24,48,and 72 h culture.The apoptosis of hPDLSCs cultured for 72 h was detected by flow cytometry.After inducing osteogenic differentiation for 21 days,alizarin red staining was used to detect the osteogenic differentiation ability of hPDLSCs.The alkaline phosphatase(ALP)activity in hPDLSCs was determined using a kit.The levels of inflammatory factors[tumor necrosis factor(TNF)-α,interleukin(IL)-1β,and IL-6]in the supernatant of hPDLSCs culture were detected by enzyme-linked immu-nosorbent assay.The mRNA expressions of osteogenic differentiation genes[Runt-associated transcription factor 2(RUNX2),osteocalcin(OCN),osteopontin(OPN)],SDF-1 and CXCR4 in hPDLSCs were detected by real-time fluores-cence quantitative polymerase chain reaction(RT-qPCR).Western blot analysis was used to determine SDF-1 and CX-CR4 protein expression in hPDLSCs.Results Compared with the blank group,the proliferative activity,number of mineralized nodules,staining intensity,ALP activity,and RUNX2,OCN,OPN mRNA,SDF-1,and CXCR4 mRNA and protein expression levels of hPDLSCs in the control group significantly decreased.The apoptosis rate and levels of TNF-α,IL-1β,and IL-6 significantly increased(P<0.05).Compared with the control group,the proliferative activity,number of mineralized nodule,staining intensity,ALP activity,and RUNX2,OCN,OPN mRNA,SDF-1,and CXCR4 mRNA and protein expression levels of hPDLSCs in low-,medium-,and high-concentration sitagliptin groups in-creased.The apoptosis rate and levels of TNF-α,IL-1β,and IL-6 decreased(P<0.05).AMD3100 partially reversed the effect of high-concentration sitagliptin on LPS-induced hPDLSCs(P<0.05).Conclusion Sitagliptin may promote the proliferation and osteogenic differentiation of hPDLSCs in LPS-induced inflammatory microenvironment by activating the SDF-1/CXCR4 signaling pathway.Furthermore,it inhibited the apoptosis and inflammatory response of hPDLSCs.

Objective To investigate the value of non-contrast CT(NCCT)-based radiomics for identifying acute unilateral middle cerebral artery occlusion(MCAO)with negative hyperdense artery sign(HAS).Methods All 80 patients with acute unilateral MCAO confirmed by angiography(MR angiography[MRA]or CT angiography[CTA]or DSA)and presenting with negative NCCT presentation for HAS were enrolled from January 2015 to June 2023 in the Emergency Department of Stroke Center of Affiliated Hospital of Yangzhou university.On the NCCT images,the occluded segment of the middle cerebral artery on the affected side of each case and the corresponding segment of the vessel on the normal side were used as the regions of interest,and a total of 108 radiomic features were extracted.The least absolute shrinkage and selection operator(LASSO)was used to screen the key features,construct and calculate the radiomics score,and four imaging histology models,support vector machine(SVM),light gradient boosting machine(LightGBM),GradientBoosting and adaptive boosting(AdaBoost),were built respectively to predict MCAO.Predictive performance was evaluated by the area under the receiver operating characteristic curves,and comparisons between the modeled receiver operating characteristic curves were made using the Delong test.Finally,the value of the application of radiological modeling was assessed by clinical decision curve analysis(DCA).Results The NCCT images based on 160 vessels were finally screened for 6 key features,including skewness,energy,gray level size zone matrix(GLSZM)-gray uneven,GLSZM-low gray area emphasis,GLSZM-size area non-uniform standardization,GLSZM-area entropy.The area under the curve(AUC)of the SVM-test was 0.688(95％CI 0.497-0.878)with an accuracy of 0.688;the AUC of the LightGBM-test was 0.787(95％CI 0.620-0.955)with an accuracy of 0.781;the AUC of the GradientBoosting-test was 0.654(95％CI 0.457-0.852)with an accuracy of 0.688;the AUC of the AdaBoost-test was 0.707(95％CI 0.515-0.899)with an accuracy of 0.750.The Delong test showed a statistically significant difference between LightGBM-test and GradientBoosting-test(P=0.040),and no statistically significant difference in performance between the remaining models(all P＞0.05).DCA showed that the LightGBM-test performed better.Conclusion NCCT-based radiomics has good diagnostic efficacy for identifying acute unilateral MCAO with negative HAS,and this conclusion needs to be further verified by multi-center and large sample studies.

According to the International League Against Epilepsy (ILAE) standards, the Newborn Working Group of the ILAE put forward 6 necessary questions about the management of neonatal anti-seizure medication and gave evidence-based recommendations in 2023. The basic framework is systematic review+expert consensus. The clinical recommendations of ILAE guidelines 2023 and the similarities and differences between ILAE guidelines 2023 and ILAE guidelines 2011 were analyzed and interpreted in this paper, in order to provide reference for colleagues involved in neonatal convulsion management in China.

Objective:To observe the effect of acupuncture combined with vitamin B12 point injection on pain severity in patients with discogenic low back pain and to analyze its potential mechanisms. Methods:A total of 96 patients with discogenic low back pain were randomly divided into two groups.The control group received acupuncture treatment,while the combined group received vitamin B12 point injection in addition to the identical acupuncture treatment in the control group.Both groups were treated for 2 weeks.The visual analog scale(VAS)and Oswestry disability index(ODI)scores were compared before treatment,after 1 week of treatment,and after 2 weeks of treatment.The levels of such serum inflammatory factors as tumor necrosis factor(TNF)-α and interleukin(IL)-6,serum beta-endorphin(β-EP),and prostaglandin(PG)E2 were compared before and after treatment.Adverse reactions and clinical efficacy were compared between the two groups after treatment. Results:The total effective rate in the combined group was higher than that in the control group(P<0.05).The VAS and ODI scores in the combined group after 1 week and 2 weeks of treatment were lower than those in the control group at the same time point(P<0.05).The levels of TNF-α,IL-6,and PGE2 in the combined group were lower than those in the control group after treatment(P<0.05),while the level of β-EP was higher than that in the control group(P<0.05).There was no statistical difference in the overall incidence of adverse reactions between the two groups(P>0.05). Conclusion:Acupuncture combined with vitamin B12 point injection can alleviate pain and promote functional recovery in patients with discogenic low back pain;reducing the levels of TNF-α,IL-6,and PGE2 and increasing the level of β-EP may be part of the mechanism of the therapy.

Humans ; Carcinoma, Non-Small-Cell Lung/therapy* ; Lung Neoplasms/pathology* ; Anaplastic Lymphoma Kinase ; Neoadjuvant Therapy ; Protein Kinase Inhibitors