Objective To explore the correlation between serum glycosylated hemoglobin(HbA1c),lym-phocyte activation gene-3(LAG-3)and thyroid nodules in patients with type 2 diabetes mellitus(T2DM).Methods A total of 120 T2DM patients with thyroid nodules admitted to the Third Hospital of Hebei Medi-cal University from July 2021 to July 2022 were included as the study group,and 100 simple T2DM patients(without thyroid nodules)were included as the control group during the same period.According to the patho-logical examination results of thyroid nodules,the study group was grouped into a benign nodule group(85 ca-ses)and a malignant nodule group(35 cases).Enzyme linked immunosorbent assay was applied to detect the serum LAG-3 level of all study subjects.Fully automated glycated hemoglobin analyzer was applied to detect HbA1c level in all study subjects.Spearman method was applied to analyze the correlation between serum HbA1c,LAG-3,and thyroid imaging report and data system(TI-RADS)scores in patients with T2DM and thyroid nodules.Multivariate Logistic regression was applied to analyze the influencing factors of T2DM with thyroid nodules.Receiver operating characteristic(ROC)curve was applied to analyze the diagnostic value of HbA1c and LAG-3 levels in T2DM with thyroid nodules.Results Compared with the control group,the level of HbA1c in the study group was obviously higher(P<0.05),while the level of LAG-3 was obviously lower(P<0.05).Compared with the benign nodule group,the serum LAG-3 level in the malignant nodule group was obviously lower(P<0.05),while the level of LAG-3 was obviously lower(P<0.05).Spearman analysis showed a positive correlation between HbA1c level and TI-RADS score in T2DM patients with thyroid nod-ules(r=0.378,P<0.001),while the serum LAG-3 level was negatively correlated with TI-RADS score(r=-0.472,P<0.001).The results of multivariate Logistic regression analysis showed that HbA1c was a risk factor for the occurrence of thyroid nodules in T2DM patients(P<0.05),and LAG-3 was a protective factor for the occurrence of thyroid nodules in T2DM patients(P<0.05).The combination of HbA1c and LAG-3 in the diagnosis of T2DM with thyroid nodules was superior to their individual diagnosis(Zcombination-HbA1c=2.542,P=0.011;Zcombination-LAG-3=3.098,P=0.002).Conclusion Patients with T2DM combined with thyroid nodules have obviously lower serum LAG-3 level and higher HbA1c level,and the two are related to the malignancy of thyroid nodules.

Objective To explore the regulatory role of hederagenin(HG)on glutamate(Glu)-in-duced ferroptosis and corresponding inflammatory responses in mouse hippocampal neuron HT22 cells and investigate its potential mechanisms.Methods HT22 cells were randomly divided into control,Glu and HG groups(n=3).The cells of the control group received no treatment,the cells of the Glu group were treated with 35 mmol/L Glu for 24 h to establish a cellular model of ferroptosis in Alzheimer's disease,and the cells of the HG group were treated with 0.5 μmol/L HG and 35 mmol/L Glu for 24 h simultaneously.FerroOrange fluorescent probe was used to de-tect intracellular Fe2+.The production of reactive oxygen species(ROS),mitochondrial membrane potential,and levels of inflammatory factors TNF-α,IL-1β and IL-6 in the cells were assessed.Finally,the expression of the key regulator of iron death,glutathione peroxidase 4(GPX4)was measured.Results Compared to the control group,the levels of intracellular Fe2+,ROS,TNF-α,IL-1β,and IL-6 were significantly elevated,while the mitochondrial membrane potential was obvi-ously reduced in the Glu group(P＜0.05,P＜0.01).The HG group had significantly decreased Fe2+,ROS,TNF-α,IL-1β,and IL-6 and enhanced mitochondrial membrane potential than the Glu group(P＜0.05,P＜0.01).The GPX4 expression was significantly lower in the Glu group than the control group(1.00±0.02 vs 0.46±0.04,P＜0.01),and was notably higher in the 0.5 and 1.0 μmol/L HG groups when compared to the Glu group(0.64±0.03 and 0.59±0.05 vs 0.46±0.04,P＜0.01).Conclusion HG inhibits ferroptosis by regulating GPX4 expression,and thereby effec-tively alleviates the inflammatory response.

Objective:To discuss the inhibitory effect of diosmetin(DIO)on the ferroptosis induced by the glutathione peroxidase(GSH-Px)inhibitor(1S,3R)-RSL3(RSL3)in spermatocytes GC-2 of the mice,and to clarify the mechanism.Methods:The GC-2 cells were divided into control group,RSL3 group,RSL3+0.8 nmol·L-1 DIO group,RSL3+4.0 nmol·L-1 DIO group,RSL3+20.0 nmol·L-1 DIO group,and RSL3+ferroptosis inhibitor Ferrostain-1(Fer-1)group(200 nmol·L-1 Fer-1).The cells were treated with 0,1,5,10,50,100,500,and 1 000 nmol·L-1 RSL3 solutions,and 0,0.5,0.1,1.0,5.0,10.0,and 50.0 μmol·L-1 DIO solutions,respectively.Additionally,the GC-2 cells were divided into blank group,model group,and treatment group.The GC-2 cells in treatment group were further divided into 0.8,4.0,and 20.0 nmol·L-1 DIO groups,as well as RSL3+0.8 nmol·L-1 DIO group,RSL3+4.0 nmol·L-1 DIO group,and RSL3+20.0 nmol·L-1 DIO group.MTT method was used to detect the survival rates of the GC-2 cells in various groups.The GC-2 cells were treated with 100 nmol·L-1 RSL3 for 0,6,12,24,36,and 48 h;Western blotting method was used to detect the expression levels of ferroptosis-related proteins in the GC-2 cells in various groups;kits were used to detect the activities of superoxide dismutase(SOD),levels of malondialdehyde(MDA),and ratios of glutathione(GSH)to glutathione disulfide(GSSG)in the GC-2 cells in various groups;immunofluorescence method was used to detect the fluorescence intensities of acyl-CoA synthetase long-chain family member 4(ACSL4)protein in the GC-2 cells in various groups.Results:The MTT method results showed that compared with 0 nmol·L-1 RSL3 group,the survival rates of the GC-2 cells in 50,100,500,and 1 000 nmol·L-1 RSL3 groups were significantly decreased(P<0.01);compared with 0 μmol·L-1 DIO group,the survival rates of the GC-2 cells in 0.5,1.0,5.0,10.0,and 50.0 μmol·L-1 DIO groups were significantly decreased(P<0.01),and 100 nmol·L-1 RSL3 with DIO concentration<0.1 μmol·L-1 were selected for the subsequent experiments.Compared with blank group,the survival rates of the GC-2 cells in model group was significantly decreased(P<0.01);compared with model group,the survival rates of the GC-2 cells in RSL3+20.0 nmol·L-1 DIO group was significantly increased(P<0.01).The Western blotting results showed that compared with 0 h,the expression level of GPX4 protein in the GC-2 cells was significantly decreased after treated with RSL3 for 6 h(P<0.01),and the expression level of HO-1 protein was significantly increased after treated with RSL3 for 12 h(P<0.05);after treated with RSL3 for 12 h,the expression levels of GPX4 and FTH1 proteins were significantly decreased(P<0.05 or P<0.01);after treated with RSL3 for 24 h,the expression levels of GPX4 and HO-1 proteins were significantly decreased(P<0.05 or P<0.01);after treated with RSL3 for 36 and 48 h,the expression levels of HO-1 protein were significantly decreased(P<0.01).Therefore,100 nmol·L-1 RSL3 and for 12 h were selected as the experimental condition for the subsequent experiments.Compared with control group,the MDA level in the GC-2 cells in RSL3 group was significantly increased(P<0.01),and the SOD activity and GSH/GSSG ratio were significantly decreased(P<0.05).Compared with RSL3 group,the SOD activities in the cells in RSL3+0.8 nmol·L-1 DIO group,RSL3+4.0 nmol·L-1 DIO group,RSL3+20.0 nmol·L-1 DIO group,and RSL3+Fer-1 group were significantly increased(P<0.05 or P<0.01).The MDA levels in the cells in RSL3+20.0 nmol·L-1 DIO group and RSL3+Fer-1 group were significantly decreased(P<0.05 or P<0.01),and the GSH/GSSG ratio in the cells in RSL3+4.0 nmol·L-1 DIO group,RSL3+20.0 nmol·L-1 DIO group,and RSL3+Fer-1 group were significantly increased(P<0.05 or P<0.01).The immunofluorescence observation results showed that compared with control group,the fluorescence intensity of ACSL4 protein in the GC-2 cells in RSL3 group was significantly increased;compared with RSL3 group,the fluorescence intensities of ACSL4 protein in the cells in RSL3+0.8 nmol·L-1 DIO group,RSL3+4.0 nmol·L-1 DIO group,RSL3+20.0 nmol·L-1 DIO group,and RSL3+Fer-1 group were significantly decreased.The Western blotting results showed that compared with control group,the expression level of HO-1 protein in the cells in RSL3 group was increased(P<0.05),and the expression levels of GPX4 and FTH1 proteins were significantly decreased(P<0.05 or P<0.01);compared with RSL3 group,the expression levels of HO-1 protein in the cells in RSL3+0.8 nmol·L-1 DIO group,RSL3+4.0 nmol·L-1 DIO group,RSL3+20.0 nmol·L-1 DIO group,and RSL3+Fer-1 group were significantly decreased(P<0.05 or P<0.01),and the expression levels of GPX4 and FTH1 proteins were significantly increased(P<0.05 or P<0.01).Conclusion:DIO can alleviate the RSL3-induced ferroptosis in the GC-2 spermatocytes of the mice,and its mechanism may be related to the inhibition of HO-1 protein expression and the upregulation of expressions of GPX4 and FTH1 proteins.

Objective To understand the changing distribution and antimicrobial resistance profiles of Proteus,Morganella and Providencia in hospitals across China from January 1,2015 to December 31,2021 in the CHINET Antimicrobial Resistance Surveillance Program.Methods Antimicrobial susceptibility testing was carried out following the unified CHINET protocol.The results were interpreted in accordance with the breakpoints in the 2021 Clinical & Laboratory Standards Institute(CLSI)M100(31 st Edition).Results A total of 32 433 Enterobacterales strains were isolated during the 7-year period,including 24 160 strains of Proteus,6 704 strains of Morganella,and 1 569 strains of Providencia.The overall number of these Enterobacterales isolates increased significantly over the 7-year period.The top 3 specimen source of these strains were urine,lower respiratory tract specimens,and wound secretions.Proteus,Morganella,and Providencia isolates showed lower resistance rates to amikacin,meropenem,cefoxitin,cefepime,cefoperazone-sulbactam,and piperacillin-tazobactam.For most of the antibiotics tested,less than 10％of the Proteus and Morganella strains were resistant,while less than 20％of the Providencia strains were resistant.The prevalence of carbapenem-resistant Enterobacterales(CRE)was 1.4％in Proteus isolates,1.9％in Morganella isolates,and 15.6％in Providencia isolates.Conclusions The overall number of clinical isolates of Proteus,Morganella and Providencia increased significantly in the 7-year period from 2015 to 2021.The prevalence of CRE strains also increased.More attention should be paid to antimicrobial resistance surveillance and rational antibiotic use so as to prevent the emergence and increase of antimicrobial resistance.

Objective:To quantitatively analyze the protein expression changes of the optic nerve in an SD rat model of non-arteritic anterior ischemic optic neuropathy (NAION), and to make bioinformatics analysis of the differential proteins.Methods:Ten 8-week-old SPF male SD rats with a body mass of 200-250 g were selected.The NAION model was established using the method of rose bengal and laser photodynamics.Four from the 8 rats with successful model were selected as the NAION model group.Another 4 body weight- and age-matched healthy SD rats without eye diseases were taken as the normal control group.The optic nerve was dissected on the 7th day after modeling.The samples were prepared by the enzyme digestion method, and the proteins were identified and quantitatively detected by isobaric tag for relative and absolute quantification labeling combined with liquid chromatography-tandem mass spectrometry.The proteins with expression fold greater than 1.5 times and significant differences between the two groups ( P<0.05) were defined as differentially expressed proteins and analyzed by bioinformatics.The use and care of animals complied with Regulations for the Administration of Affair Concerning Experimental Animals by the State Science and Technology Commission of China.The study protocol was approved by an Animal Ethical and Welfare Committee of Tianjin Medical University Eye Hospital (No.TJYY2021041029). Results:Three days after modeling, the optic disc of rats was swollen and fluorescein leakage in the optic disc was detected in fluorescein fundus angiography images in the NAION model group, which indicated the model was established successfully.A total of 1 291 quantifiable proteins including 80 differentially expressed proteins were identified.Compared with the normal control group, there were 5 up-regulated proteins and 75 down-regulated proteins.The expression levels of collagen alpha-1(V) chain (Col5A1), cAMP-dependent protein kinase catalytic subunit beta (Prkacb) and disks large homolog 1(Dlg1) were increased, and the expression levels of neurofilament medium polypeptide (Nefm), microtubule-associated protein 1B (Map1b), Ras-related protein Ral-A (Rala), serine/threonine-protein kinase N2 (Pkn2) and platelet-activating factor acetylhydrolase IB subunit beta (Pafah1b1) were decreased.Differentially expressed proteins were mainly involved in the biological processes, including regulation of the cytoskeleton, cellular response to hypoxia, axon production and extension, regulation of synapse, regulation of neuron apoptosis and axo-dendritic transport, etc.KEGG pathway enrichment analysis showed that differentially expressed proteins were mainly involved in metabolic pathways, synaptic vesicle circulation and platelet activation.Conclusions:The expression of proteins related to signal pathways such as nerve growth, energy metabolism, axo-dendritic transport and apoptosis is involved in the apoptosis of neurons in NAION.

Hepatitis B virus (HBV) infection is a major global public health issue. Clinical cure (also known as functional cure) of chronic hepatitis B (CHB) is the ideal therapeutic goal recommended by the latest guidelines for the prevention and treatment of CHB in China and globally. Optimized treatment regimens with direct-acting antiviral agents ［e.g., nucleos(t)ide analogues］ or immunomodulators (e.g., pegylated interferon-α) sequentially or in combination tend to have low cure rates. Rapid development has been achieved in the research and development of drugs for the treatment of CHB. This article reviews the clinical study of new antiviral drugs for CHB, including the selection of subjects, study design, dosage, dose escalation, adverse events, and efficacy evaluation. It is necessary to introduce the knowledge of quantitative pharmacology to analyze the association of drug exposure in body with efficacy and adverse reactions, and exploratory indicators should be incorporated for comprehensive analysis. This review provides related experience and new ideas for the clinical research and development of new anti-HBV drugs.

Objective:To analyze the protein expression changes in the retina of non-arteritic anterior ischemic optic neuropathy (NAION) in rats.Methods:The rat NAION (rNAION) model was established by Rose Bengal and laser. Twenty Sprague-Dawley rats were randomly divided into 4 groups, the normal control group, the laser control group, the RB injection control group, and the rNAION model group, with 5 rats in each group. The right eye was used as the experimental eye. The retina was dissected at the third day after modeling. Enzyme digestion method was used for sample preparation and data collection was performed in a non-dependent collection mode. The data were quantitatively analyzed by SWATH quantitative mass spectrometry, searching for differential proteins and performing function and pathway analysis.Results:Compared with the other three control groups, a total of 184 differential proteins were detected in the rNAION group (expression fold greater than 1.5 times and P<0.05), including 99 up-regulated proteins and 85 down-regulated proteins. The expressions of glial fibrillary acidic protein, guanine nucleotide binding protein 4, laminin 1, 14-3-3γ protein YWHAG were increased. Whereas the expressions of Leucine-rich glioma-inactivated protein 1, secretory carrier-associated membrane protein 5, and Clathrin coat assembly protein AP180 were decreased. The differential proteins are mainly involved in biological processes such as nerve growth, energy metabolism, vesicle-mediated transport, the regulation of synaptic plasticity, apoptosis and inflammation. Pathway enrichment analysis showed that PI3K-Akt signaling pathway and complement and thrombin reaction pathway was related to the disease. Conclusion:The protein expressions of energy metabolism, nerve growth, synaptic vesicle transport and PI3K-Akt signaling pathway can regulate the neuronal regeneration and apoptosis in NAION.

Objective:To validate the role of color Doppler ultrasound in an animal model to detect early heterotopic ossification (HO) after brain-traumatic/burn/tenotomy.Methods:Forty-four rats were randomly divided into two groups. Rats in experimental group ( n=22) were operated to build brain-traumatic/burn/tenotomy model and others in control group ( n=22) underwent only skin incision injury. Color Doppler ultrasound, X-ray film examination at 2, 3, 4, 6, 8 and 10 weeks post-injury were performed to follow up the progression of HO in both groups respectively. Histology was used to confirm bone formation. Results:In the experimental group, disorder structure with a hypoechoiccore in treated Achilles tendon was visualized using color Doppler ultrasound in the 2nd week. Additional tiny hyperechoic foci were observed in the 3rd week, which increased in the fourth week and fused into a mineralized island in the sixth week. No obvious abnormality was found in control group at the aforementioned time point. X-ray could detect heterotopic bone tissue in the sixth week in the experimental group but not in the control group. X-ray and HE stainning had confirmed bone formation in the tenth week in the experimental group.Conclusions:Color Doppler ultrasound can detect early HO and continuously follow up the progression of HO.

Esophageal squamous cell carcinoma (ESCC) is a deadly malignancy with regard to mortality and prognosis, and the 5-year survival rate for all patients diagnosed with ESCC remains poor. A better understanding of the biological mechanisms of ESCC tumorigenesis and progression is of great importance to improve treatment of this disease. In this study, we demonstrated that the glutathione metabolism pathway is highly enriched in ESCC cells compared with normal esophageal epithelial cells in an in vivo mouse model. In addition, treatment with L-buthionine-sulfoximine (BSO) to deplete glutathione decreased the ESCC tumor burden in mice, thus demonstrating the critical role of glutathione metabolism in ESCC progression. BSO treatment also led to decreased cell proliferation and activation of cell apoptosis in ESCC. Finally, BSO treatment blocked NF-κB pathway activation in ESCC. Our study reveals a new pathway that regulates ESCC progression and suggests that inhibition of glutathione metabolism may be a potential strategy for ESCC treatment.
Animals ; Apoptosis ; Carcinogenesis ; Carcinoma, Squamous Cell ; Cell Proliferation ; Epithelial Cells ; Esophageal Neoplasms* ; Glutathione* ; Humans ; Metabolism* ; Mice ; Mortality ; Prognosis ; Survival Rate ; Tumor Burden

Objective To investigate the prevalence of urine abnormalities for school children in Chengdu city and to evaluate the significance of urinary screening.Methods During January to December 2013,morning urine of 6 615 students were collected and screened by urine reagent paper.Two weeks later,the repeated screening was conducted in the children whose urine samples were positive for the first screening.Urine samples with positive testing results for twice were submitted to urine routine tests at local hospital,and the children with the urine positive results were defined as urine abnormalities.The children with urine abnormalities were transferred to a tertiary hospital and given treatment and follow-up.Results There were 6 615 cases receiving urine screening,including 2 624 cases (39.67 ％) of the grade I,and 3 991 cases(60.33％) at junior middle school.During the first screening,323 cases (4.83％) children had urinary occult blood positive,43 cases (0.65％) had urinary protein,20 cases (0.30％) had occult blood positive and proteinuria,and 103 cases (1.56％) had white cells in urine.During the second urine screening,62 cases (0.94％) had occult blood positive,6 cases (0.09％) had urinary protein,2 cases (0.03％) had proteinuria and occult blood positive,46 cases (0.70％) had white cells in urine.The incidence of urine abnormalities with occult blood positive,proteinuria,occult blood positive and proteinuria,and white cells in urine of children at junior middle school [1.38％ (55/3 991 cases),0.13％ (5/3 991 cases),0.05％ (2/3 991 cases),0.70％ (28/3 991 cases)] were significantly higher than those of children at primary school [0.27％ (7/2 624 cases),0.04％ (1/2 624 cases),0 (0/ 2 624 cases),0.69％ (18/2 624 cases)],and all the differences were statisticallysignificant (x2 =64.16,168.53,178.09,98.16,all P ＜ 0.05).In children transferred to a tertiary hospital for treatment,there were 4 cases with IgA nephropathy,1 case with minor glomerular abnormalities,and 12 cases with urinary tract infection.Conclusion Urinary screening is an effective way to find out kidney disease and urinary tract infection in children.Follow-ups should be strengthened.