Objective:To preliminarily explore the role of silent information regulator 3 (SIRT3) in ferroptosis induced by high glucose in renal tubular epithelial cells, and to provide a new theoretical basis and treatment ideas for renal tubular injury in diabetic kidney disease patients.Methods:The single-cell transcriptomic analysis from "Tabula-muris" database was used to evaluate the expression of SIRT3 gene in different cellular subtypes of kidney tissues. HK-2 cells, a human immortalized proximal tubule epithelial cell line, were cultured in vitro and divided into following groups: (1) control group, mannitol group and high glucose group; (2) control group, negative control group, SIRT3 overexpression group, high glucose group and SIRT3 overexpression + high glucose group; (3) control group, negative control group, SIRT3 knockdown group, high glucose group and SIRT3 knockdown + high glucose group; (4) control group, Erastin intervention group and SIRT3 overexpression + Erastin intervention group. Normal glucose was 5.5 mmol/L, high glucose was 30 mmol/L, mannitol was 24.5 mmol/L, Erastin was 10 μmol/L, and the intervention time was 48 h. Cell counting kit-8 proliferation and cytotoxicity assay was used to determine cell viability. Real-time quantitative PCR and Western blotting were performed to assess the expression of SIRT3, kidney injury molecule-1 (KIM-1), and ferroptosis-related proteins acyl-CoA synthetase long chain family member 4 (ACSL4) and glutathione peroxidase 4 (GPX4) at the mRNA and protein levels. The malondialdehyde, glutathione, and iron levels were measured to evaluate the degree of cellular ferroptosis. DCFH-DA was used to analyze the intracellular reactive oxygen species level, while the JC-1 staining method was employed to evaluate alterations of mitochondrial membrane potential in HK-2 cells. Results:(1) The results of single-cell transcriptomic database analysis demonstrated that SIRT3 gene was expressed at the highest level in the subtypes of proximal tubule epithelial cells of kidney tissues. (2) Compared with the control group, the expression levels of KIM-1 and ACSL4 were higher, and the expression levels of SIRT3 and GPX4 and cell viability were lower in the high glucose group (all P<0.05), while there was no statistically significant difference of the aforementioned indicators between the mannitol group and the control group (all P>0.05). (3) Compared with the high glucose group, HK-2 cell vitality, GPX4 expression and intracellular glutathione were higher, ACSL4 expression, intracellular iron, malondialdehyde and reactive oxygen species were lower, mitochondrial membrane potential partially recovered in SIRT3 overexpression + high glucose group (all P<0.05). Compared with the high glucose group, HK-2 cell vitality and GPX4 expression were lower, ACSL4 expression was higher in SIRT3 knockdown + high glucose group (all P<0.05), and there were no statistically significant differences in intracellular iron, malondialdehyde and glutathione (all P>0.05). (4) Compared with the control group, Erastin intervention group had upregulated ACSL4 expression and downregulated GPX4 expression in HK-2 cells (all P<0.05). Compared with the Erastin intervention group, SIRT3 overexpression + Erastin intervention group had upregulated GPX4 expression and downregulated ACSL4 expression (all P<0.05). Conclusions:High glucose can decrease SIRT3 expression and mitochondrial membrane potential, and increase oxidative stress and ferroptosis in HK-2 cells. Overexpression of SIRT3 may reduce oxidative stress and alleviate mitochondrial dysfunction, thereby mitigating glucose-induced ferroptosis in HK-2 cells.

AIM:To investigate the effects of γ-synuclein on autophagy and apoptosis of colon cancer cells in-duced by endoplasmic reticulum stress,as well as the protective effect on the cells.METHODS:Gene expression profile chip analysis was performed to compare the cDNA expression profiles between human colon cancer HCT116 cells with γ-synu-clein knockdown and HCT116 cells with control siRNA,and to identify potential molecules related to autophagy and apop-tosis.In colon cancer cell lines,the functional effects of γ-synuclein on autophagy and apoptosis induced by thapsigargin(TG),an endoplasmic reticulum stress-inducing agent,were systematically explored by conducting immunofluorescence staining,Western blot,CCK-8 assay,flow cytometry,and transmission electron microscopy.Western blot was used to de-tect the expression of γ-synuclein protein,autophagy-related proteins[microtubule-associated protein 1 light chain 3(LC3),beclin-1,autophagy-related protein 5(ATG5)and ATG7],and apoptosis-related proteins[poly(ADP-ribose)polymerase(PARP),pro-caspase-3,and pro-caspase-9].To further analyze the mechanism of γ-synuclein in regulating autophagy and apoptosis,extracellular signal-regulated kinase(ERK)inhibitor PD98059,ERK inhibitor SP600125 and c-Jun N-terminal kinase(JNK)activator anisomycin were applied separately to test HCT116 cells transfected with γ-synu-clein siRNA.Subsequently,autophagy proteins,apoptosis proteins,and ERK and JNK pathway-related proteins were de-tected by Western blot.RESULTS:The TG-induced autophagy of colon cancer cells mainly occurred at the early stage(0～24 h),and apoptosis mainly occurred at the late stage(36～48 h).Endoplasmic reticulum stress up-regulated the ex-pression of γ-synuclein in colon cancer cells,which was associated with enhanced autophagy.γ-Synuclein promoted au-tophagy by activating ERK and JNK pathways at the early stage(0～24 h),and inhibited apoptosis by blocking JNK path-ways at the late stage(24～48 h)to protect HCT116 cells.In our model,γ-synuclein was observed to play a critical role in the transition from endoplasmic reticulum stress-induced autophagy to apoptosis.CONCLUSION:In the context of endo-plasmic reticulum stress,γ-synuclein promotes autophagy and inhibits apoptosis by regulating ERK and JNK signaling pathways,thus protecting colon cancer cells.This provides a potential idea for anti-tumor therapy.

Objective:To discuss their effects of PLA(polylactic acid)/PTMC(polytrimethylene carbonate)and PLA/PTMC-calcium phosphate[Ca3(PO4)2]composite porous scaffolds prepared by 3D printing technology on bone marrow mesenchymal stem cells(BMSCs)of the rabbits,and to clarify their application values in bone defect repairment.Methods:After mixing the materials,PLA/PTMC and PLA/PTMC-Ca3(PO4)2 filaments were prepared by desktop filament extruder.The scaffolds were designed by CATIA V5-6R2019 modeling software and fabricated using CreatBot F430 3D printer.The chemical structure of the PLA/PTMC-Ca3(PO4)2 scaffold was detected by infrared spectroscopy.In vitro degradation experiments were used to detect the degradation weight loss rates and pH values of the two scaffolds.A contact angle measuring instrument was used to detect the hydrophilicities of the two scaffolds.The BMSCs were extracted from three newborn New Zealand white rabbits(2-5-day-old);CCK-8 method was used to detect the proliferation activities of the cells co-cultured with two scaffolds,and Alizarin red staining was used to observe the osteogenic differentiation of the cells co-cultured with two scaffolds.Results:Infrared spectroscopy confirmed the successful preparation of composite scaffolds containing PLA,PTMC,and β-Ca3(PO4)2.During degradation for 6-14 weeks,compared with PLA/PTMC scaffold,the degradation rates of the PLA/PTMC-Ca3(PO4)2 scaffold in lipase solution and phosphate-buffered saline(PBS)were significantly increased(P<0.05 or P<0.01).During degradation for 8-14 weeks,compared with PLA/PTMC scaffold,the pH value of the PLA/PTMC-Ca3(PO4)2scaffold in lipase solution was significantly increased(P<0.01).Compared with PLA/PTMC scaffold,the contact angle of the PLA/PTMC-Ca3(PO4)2 scaffold was significantly decreased(P<0.01).On days 5 and 7 of cell co-culture,compared with PLA/PTMC scaffold,the proliferation activity of the cells co-cultured with PLA/PTMC-Ca3(PO4)2 scaffold was significantly increased(P<0.05 or P<0.01).After 21 d of co-culture,both scaffolds overlapped with BMSCs and locally formed calcified nodules,which were stained orange by Alizarin red.Compared with PLA/PTMC scaffold,the number of mineralized calcium nodules in the cells co-cultured with PLA/PTMC-Ca3(PO4)2 scaffold was increased,with greater density and deeper color.Conclusion:The PLA/PTMC-Ca3(PO4)2 composite porous scaffolds containing PLA,PTMC,and β-Ca3(PO4)2 are successfully prepared by 3D printing technology.These scaffolds exhibit good degradation properties and show advantages in biocompatibility,hydrophilicity,and osteogenic induction;they are excellent materials for the bone defect repairment.

Objective To investigate the research status and the research hotspots on male nurses at home and abroad in last decade,so as to provide references for further research in male nurses.Methods Literature between 2013 and 2024 on male nurses were retrieved from the Web of Science(WOS)and CNKI.CiteSpace 6.2.6 R7 software was used to analyse the volume of publication,keyword co-occurrence,keyword clustering and emergence.Results A total of 736 articles were included in the study.From 2013 to 2023,the number of published papers of male nurses in CNKI and WOS showed an increasing trend.The research was primarily affiliated with 263 institutions across 62 countries.The country with the highest volume of publication is China,followed by the United States,although there was a need to enhance collaboration with other nations.The institution with the most publications is the Hong Kong Polytechnic University.A comprehensive analysis of keyword co-occurrence,clustering,and emerging trends revealed that the research hotspots in the field of male nursing in CNKI mainly includes the advantages of male nurses,psychological capital,humanistic care,development,needs,professional identification.Whereas WOS focuses on the topics such as gender bias against male nurses,mental health,workplace violence,ambivalent sexism,motivational barriers,individual differences,societal prejudice and knowledge and attitucle.Both domestic and international research on male nurses primarily consist of cross-sectional surveys and qualitative studies,with fewer intervention studies.Currently,the researches regarding male nursing focus on the qualitative study of mental health of male nurses.Conclusion Focusing on the mental health and professional status of male nurses,domestic scholars need to increase the formulation and implementation of interventions in this field in order to address the practical issues faced by male nurses,therefore to promote the comprehensive development of the nursing profession,enhance the overall quality and service quality of the nursing team,and provide patients with higher quality in nursing services.

Objective:To establish a multi-factor prediction model for vascular aging based on psychological behavior and physical exercise using nomogram and structural equation method.Methods:A total of 701 inpatients or physical examination patients who underwent carotid ultrasound examination in the Affiliated Hospital of Qingdao University from October 2021 to March 2023 were selected by cross-sectional method. The subjects were randomly divided into training queue( n=492) and verification queue( n=209) according to the ratio of 7∶3.The psychological and behavioral factors, lifestyle, demography and accompanying diseases data were collected. Multi-factor binary Logistic regression method in SPSS 25.0 software was used to screen independent influencing factors of vascular aging, R 4.2.2 software was used to build a nomogram prediction model of vascular aging, and the ROC curve, calibration chart and decision curve analysis were used to evaluate the nomogram, and AMOS software was used to build a structural equation model of vascular aging. Results:Multivariate Logistic regression analysis showed that type A behavior( B=1.757, OR(95% CI)=5.790(2.750-12.210), P<0.001), physical exercise( B=-3.019, OR(95% CI)=0.050(0.020-0.100), P<0.001), high-fat diet( B=0.679, OR(95% CI)=1.970(1.350-2.880), P<0.001), sleep quality( B=-1.451, OR(95% CI)=0.230(0.120-0.460), P<0.001), tea drinking habits( B=-2.349, OR(95% CI)=0.100(0.050-0.200), P<0.001), age( B=0.061, OR(95% CI)=1.060(1.020-1.110), P=0.005), sex( B=-1.263, OR(95% CI)=0.280(0.140-0.570), P<0.001), hyperlipidemia( B=0.679, OR(95% CI)=1.970(1.350-2.880), P<0.001) and diabetes( B=0.838, OR(95% CI)=2.310(1.140-4.700), P=0.021)were independent influencing factors of vascular aging. The nomogram model showed that type A behavior, physical exercise, high-fat diet, sleep quality, tea drinking habits, age, sex, hyperlipidemia and diabetes were independent influencing factors of vascular aging.Advanced age, type A behavior, hyperlipidemia, diabetes and the score of high-fat diet were risk factors for vascular aging, while better sleep quality, tea drinking habit and the score of physical exercise were protective factors for vascular aging.Men were more prone to vascular aging than women. Structural equation model showed that type A behavior had the strongest direct positive effect on vascular aging (the effect coefficient=0.197, P<0.01), and physical exercise had the strongest direct negative effect on vascular aging (the effect coefficient=-0.452, P<0.01), which could indirectly affect vascular aging through various factors. Conclusions:The nomogram model shows the independent influencing factors of vascular aging and has certain predictive value.Structural equation model shows that many factors such as psychological behavior and lifestyle can directly or indirectly affect the occurrence of vascular aging, among which type A behavior and physical exercise have more extensive effects.

Abstract OBJECTIVE To establish an efficient and simple HPLC-MS/MS method for determination of flucloxacillin in newborn plasma, and to investigate the interaction between ambroxol and flucloxacillin in newborns. METHODS The samples were analyzed by API4000 HPLC-MS/MS. Ultimate XB-C18 column(2.1 mm×100 mm, 5 μm) were carried out. The mobile phase was composed of water-0.1% formic acid(A) and acetonitrile-0.1% formic acid(B). The quantitative analysis of the ion transitions were monitored at m/z 452.6→284.2 for flucloxacillin and m/z 821.4→397.3 for rifampicin(internal standard). RESULTS The linear range of flucloxacillin under this analysis method was 0.20-80 ng·mL-1, and the lower limit of quantification was 0.20 ng·mL-1; The intra-day and inter-day precision of flucloxacillin were both less than 8.23%; The extraction recovery was in the range of 85.3%-89.2%, and the matrix effect was between 89.3%-92.3%; The stability of plasma samples was good under conditions of 12 h at room temperature, 4 h at room temperature after treatment, repeated freeze-thaw for 3 times, and -20 ℃ freezing for 30 d. The results of clinical samples indicated that the combination of ambroxol could significantly increase the blood concentration of flucloxacillin. CONCLUSION The established HPLC-MS/MS method is accurate, sensitive and can be used for the determination of flucloxacillin concentration in neonatal plasma. The results of clinical samples indicate that ambroxol can significantly increase the blood concentration of flucloxacillin. There are drug interactions between ambroxol and flucloxacillin.

One of the distinct hallmarks of cancer cells is aerobic glycolysis (Warburg effect). Lactate dehydrogenase A (LDHA) is thought to play a key role in aerobic glycolysis and has been extensively studied, while lactate dehydrogenase C (LDHC), an isoform of LDHA, has received much less attention. Here we showed that human LDHC was significantly expressed in lung cancer tissues, overexpression of Ldhc in mice could promote tumor growth, and knock-down of LDHC could inhibit the proliferation of lung cancer A549 cells. We solved the first crystal structure of human LDHC4 and found that the active-site loop of LDHC4 adopted a distinct conformation compared to LDHA4 and lactate dehydrogenase B4 (LDHB4). Moreover, we found that (ethylamino) (oxo)acetic acid shows about 10 times selective inhibition against LDHC4 over LDHA4 and LDHB4. Our studies suggest that LDHC4 is a potential target for anticancer drug discovery and (ethylamino) (oxo)acetic acid provides a good start to develop lead compounds for selective drugs targeting LDHC4.

Objective:To analyze the effect of spine-pelvis sagittal parameters and sagittal orientation of facet joint on degeneration of cranial L 3,4 facet joint (facet joint degeneration, FJD) after L 4-S 1 posterior lumbar interbody fusion (PLIF). Methods:Patients with lumbar degenerative diseases who underwent L 4-S 1 PLIF from January 2012 to December 2016 were retrospectively investigated, there were 54 cases, including 28 males and 26 females. Age: 54.59±5.48 years (range, 45-60 years). X-ray, CT, MRI and Weishuapt grade was used to evaluate the degeneration of L 3,4 facet joint at the cranial adjacent segment. The general information and the sagittal parameters of spine pelvis at the last follow-up were compared between the two groups. The former included age, gender, body mass index (BMI), bone mineral density (BMD), follow-up time and preoperative diagnosis. The latter included lower lumbar lordosis angle (LLL), lumbar lordosis angle (LL), pelvis incidence (PI), pelvis tilt (PT), sacrum slope (SS), the height of the intervertebral space (HD), the angle of cranial facet joint, Oswestry disability index (ODI), Japanese Orthopedic Association (JOA) lumbar function score and improvement rate were compared at the same time. Independent sample t-test was used to compare continuous variables between groups; comparison of categorical variable components χ 2 test or Fisher's exact test. Multivariate logistic regression analysis was used to predict the risk factors of adjacent FJD. Results:Postoperative follow-up was 33.44±6.85 months (range, 24-36 months), there were 17 patients in the degenerative group and 37 patients in the non degenerative group. There were no significant differences in age, gender, BMI, BMD, follow-up time or preoperative diagnosis between the two groups. LLL, LL and SS also showed no significant difference. At the last follow-up, PI (56.28°±6.03° vs. 47.87°±8.30°, t=3.74, P=0.001), PT (17.90°±7.06° vs. 14.41°±5.51°, t=1.97, P=0.042) and the joint angle of the cephalic facet (58.48°±2.00° vs. 54.69°±3.01°, t=4.72, P=0.072) in the degenerative group were greater than those in the non-degenerative group. In the subgroup analysis of lumbar lordosis distribution, the difference between the two groups was statistically significant (χ 2=9.90, P=0.006). The HD in the degenerative group 7.50±3.60 mm was significantly lower than that in the non degenerative group 9.30±2.79 mm ( t=2.00, P=0.031). Multivariate logistic regression analysis showed that increase of PI ( OR=1.22, P=0.005) and magnified cephalic facet joint angle ( OR=2.04, P=0.008) were risk factors for adjacent segment facet degeneration. At the last follow-up, the ODI improvement rate in the degenerative group (58.14%±13.41% vs. 70.18%±8.03%, t=4.11, P<0.001) and the JOA score improvement rate (44.72%±9.53% vs. 68.86%±8.55%, t=0.43, P=0.001) were lower than those in the non degenerative group. Conclusion:The increase of PI and sagittal facet (increased joint angle of proximal facet) are risk factors of adjacent segment FJD after lumbar fusion; The abnormal distribution of lower lumbar lordosis and poor PT recovery in adjacent segment FJD patients after lumbar fusion are more obvious, which may be related to the increase of PI; After lumbar fusion, the orientation of adjacent facet joint tended to be sagittal.

OBJECTIVE：To study the effects o f CYP2C9*3 gene polymorphism on therapeutic efficacy of benzbromarone in lowering uric acid and its hepatotoxicity. METHODS ：A retrospective study was conducted to analyze the relevant clinical indicators and genotypes of 196 gout patients who received benzbromarone and CYP2C9*3 gene polymorphism test in Wuhan third hospital from Jan. 2018 to Sept. 2019. RESULTS ：Among 196 patients，179，15 and 2 patients with CYP2C9*3 genotypes * 1/*1， *1/*3 and * 3/*3 genotypes were found ，respectively，and the distribution of each genotype was in line with Hardy-Weinberg balance（P＞0.05）. Before treatment ，there were no significant differences in the levels of UA ，Scr，ALT，AST and CRP between *1/*1 genotype and * 1/*3+*3/*3 genotype（P＞0.05）. After 4 weeks of treatment ，the UA ，Scr，CRP levels of patients with * 1/*1 genotype as well as the UA and CRP levels of patients with * 1/*3+*3/*3 genotype were significantly reduced ，the UA level of patients with * 1/*1 genotype was significantly lower than that of patients with * 1/*3+*3/*3 genotype（P＜0.05 or P＜0.01）. The ALT and AST levels had no obvious changes in patients with different genotype before and after treatment ，and they were in the normal range. No serious abnormal liver function was observed during the treatment. CONCLUSIONS ：Therapeutic efficacy of benzbromarone in lowering uric acid in gout patients with CYP2C9*3 genotypes * 1/*1 genotype is better than that of * 1/*3 and * 3/*3 genotypes. However ，the gene polymorphism may be not associated with its hepatotoxicity.

Mycoplasma pneumoniae pneumonia (MPP) complicated with cerebral venous sinus thrombosis (CVST) is rare.We retrospectively analyzed the clinical data of two patients with refractory mycoplasma pneumoniae pneumonia (RMPP) complicated with CVST who were hospitalized in Xi′an children′s Hospital from December 2018 to April 2019, inquired the relevant literature, analyzed the clinical diagnosis and treatment characteristics, and discussed the diagnosis and treatment measures of RMPP complicated with CVST.Two cases were 6-year-old girls with fever and cough as the main symptoms.After physical examination, the respiratory sounds of the affected lung decreased, and the sounds of phlegm and dampness could be heard in both lungs.Mycoplasma pneumoniae (MP) antibody and RNA were positive.Chest CT showed lobar pneumonia with a large number of pleural effusion.The effect of macrolide antibiotics anti infection treatment was not good.Headache symptoms occurred during the course of the disease, and serum D-dimer increased significantly.Brain MRI showed CVST, including 1 case with lower extremity pain, and B-ultrasound showed right lower extremity arterial embolism.After anti infection, thrombectomy, anticoagulation and symptomatic treatment, 2 cases were discharged.When children with MPP, especially those with RMPP, have extracranial thrombosis and/or neurological symptoms, accompanied by elevated serum D-dimer, the possibility of CVST should be considered, and brain MRI examination should be performed in time to confirm and actively treat, which can reduce or avoid the occurrence of sequelae.Thrombosis may be related to excessive inflammatory reaction and vascular endothelial injury caused by MP infection.