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.

ObjectiveTo reveal the pharmacodynamic substances for the anti-inflammatory and analgesic effects of Glycyrrhizae Radix et Rhizoma by structure-activity omics. MethodOn the basis of the previous study about the screening of active components in vitro， this study explored the effects of flavonoids in Glycyrrhizae Radix et Rhizoma in vivo. The flavonoids in Glycyrrhizae Radix et Rhizoma and their direct targets for the anti-inflammatory and analgesic effects were obtained from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP）， PharmMapper， Swiss TargetPrediction， DisGeNET， and Online Mendelian Inheritance in Man （OMIM）. STRING and Cytoscape 3.7.2 were employed to establish the protein-protein interaction （PPI） network of key targets. Molecular docking was performed to simulate the binding of five targets with high degrees to flavonoids in Glycyrrhizae Radix et Rhizoma， on the basis of which the key core targets were selected. The targets were used as a bridge to correlate the structures and effects of one or more classes of chemical components in Glycyrrhizae Radix et Rhizoma. According to the binding affinity between flavonoids with different structures in Glycyrrhizae Radix et Rhizoma and targets， the relationships between compound structures and core targets were discussed. ResultThe flavonoids in Glycyrrhizae Radix et Rhizoma reduced the content of prostaglandin E₂ （PGE₂） in the rat model of pain induced by formalin， demonstrating definite anti-inflammatory and analgesic effects. Sixty active compounds （flavonoids） with anti-inflammatory and analgesic effects of Glycyrrhizae Radix et Rhizoma were obtained. With the total score as the standard， prostaglandin-endoperoxide synthase 2 （PTGS2） and mitogen-activated protein kinase 3 （MAPK3） were selected as the key core targets of Glycyrrhizae Radix et Rhizoma for the anti-inflammatory and analgesic effects. Except that flavones showed selectivity of binding to MAPK3， the other flavonoids of Glycyrrhizae Radix et Rhizoma showed strong binding to PTGS2 and MAPK3， and the structures containing glycoside fragments showed stronger binding affinity to the targets. The introduction of chain olefins in the ring of chalcones facilitated the binding to the targets. The isopentenyl fragment in flavonols may cause the difference in binding affinity. The parallel combination of a ring into pyran ring in flavanes was not conducive to the binding to the target. The electric charge， liposolubility， and steric hindrance of the substituent group on the B ring of isoflavones directly affected the binding affinity. ConclusionThis study adopts structure-activity omics to analyze the material basis for the anti-inflammatory and analgesic effects of Glycyrrhizae Radix et Rhizoma. Structure-activity omics provides new ideas and methods for predicting the pharmacodynamic substances of traditional Chinese medicine.

Objective To investigate the effects of glycogen synthase kinase-3β (GSK-3β) and β-catenin signaling on the human renal proximal tubular epithelial HK-2 cell injury induced by depleted uranium(DU),and provide a new enlightenment for the development of DU antidotes.Methods H K-2 cells were exposed to different concentrations of DU for 3-24 h,then the protein expressions of kidney injury molecule 1 (KIM-1),neutrophil gelatinase-associated lipocalin (NGAL) and nuclear β-catenin were detected by immunofluorescence staining.The protein expressions of p-GSK-3 β(S9),GSK-3β and cmyc were detected by Western blot assay.HK-2 cells were transiently transfected by GSK-3β (KD) plasmid or treated by TDZD-8 to inhibit the activity of GSK-3β specifically.Other HK-2 cells were transiently transfected by β-catenin plasmid to overexpress the β-catenin protein.Results The percentages of KIM-1 and NGAL-positive cells increased with DU exposure time and concentrations from 300 and 600 μmol/L,and they were significantly higher than those of the blank control at 6-24 h of DU exposure (KIM-1-positive cells:t =11.06,18.97,30.49,P ＜0.05;t =6.79,16.02,85.45,P ＜ 0.05;NGAL-positive cells:t =11.78,11.37,34.29,P ＜0.05;t =7.34,21.63,36.84,P ＜0.05).In contrast,the ratio of p-GSK-3β (S9) to GSK-3β and percentage of nuclear β-catenin-positive cells were significantly higher than that of the blank control at 3-24 h of DU exposure (p-GSK-3β(S9)/GSK-3β:t =3.95,4.69,5.40,3.34,P ＜ 0.05;nuclear β-catenin-positive cells:t =4.61,6.52,36.64,14.93,P ＜ 0.05) with a maximum response at 9 h of DU exposure accompanied with corresponding increase of protein level of c-myc,a downstream target gene of β-catenin.Transient transfection of HK-2 cells with GSK-3β (KD) plasmid significantly inhibited the activity of GSK-3β (t =8.07,P ＜ 0.05) and reduced the DU-increased percentage of KIM-1-positive cells (t =24.77,P ＜ 0.05).Treatment cells with TDZD-8 inhibited the activity of GSK-3β and enhanced the percentage of nuclear β-catenin-positive cells,and it also significantly reduced the percentage of KIM-1-positive cells in HK-2 cells exposed to DU (t =6.25,6.73,P ＜ 0.05).Moreover,overexpression of β-catenin significantly reduced DU-induced cell injury (t =7.48,P ＜ 0.05).Conclusions GSK-3β/β-catenin signaling plays a key role in regulating the DU-induced cytotoxicity of HK-2 cells.Inhibition of GSK-3β activity and overexpression of β-catenin can protect the HK-2 cells from DU-induced damage.

Objective: To study the VP1 gene of coxsackievirus A6 (CVA6) and to reveal the molecular epidemiologic characteristics of CVA6 related to hand, foot and mouth disease (HFMD) in Anyang city from 2011 to 2015. Methods: Serotypes of human enterovirus (HEV) were detected with the real-time RT-PCR from the clinical specimens. Primers were designed and used to amply and sequence the VP1 region of CVA6. Phylogenetic tree was constructed and phylogenetic analysis was performed. Selection pressures of the VP1 gene were evaluated for different subgenotypes of CVA6 circulating in China in recent years. Results: A total of 365 specimens with CVA6 positivity were identified in Anyang city during 2011—2015. CVA6 specimens accounted for 19.59% of all HEV-positive specimens. Phylogenetic analysis showed that all CVA6 strains were divided into four genotypes and the genotype D was further divided into two subgenotypes. Among the twenty-two Anyang sequences involved in the phylogeny of CVA6, five sequences belonged to subgenotype D1 and the other seventeen sequences belonged to subgenotype D2. The values of selection pressure of the Chinese CVA6 strains within subgenotype D1 and subgenotype D2 were 0.031 and 0.075, respectively. Only one positively selected site was detected in the VP1 gene of subgenotype D1, meanwhile four were detected in the VP1 gene of subgenotype D2. Conclusions Subgenotype D1 and its successor, subgenotype D2, circulated in Anyang city during 2011—2015. The subgenotype replacement from CVA6 subgenotype D1 to CVA6 subgenotype D2 and the continuous transmission of CVA6 subgenotype D2 were the two major causes of HFMD epidemic related to CVA6 in Anyang city during 2013—2015.

Objective To predict incidence trend of brucellosis in Anyang,Henan Province,so as to provide basis for instituting aimed preventive management.Methods To construct gray model (GM) according to incidence and rate of brucellosis of Anyang from 2004 to 2015,and predict the incidence rate of brucellosis of Anyang from 2016 to 2018.The data of brucellosis in Anyang were from the Infectious Disease Report Information Management System for Disease Control,China Information System for Disease Control and Prevention,the population data were from Anyang Bureau of Statistics.Results The GM (1,1) had good accuracy in predicting and test,the posterior error ratio (C) =0.32,and the probability value (P) =0.99.The model could be used for extrapolating prediction.The actual incidence rate was compared with the predicted value,the value was basically consistent,fit effect was good.The predictive morbidity of brucellosis was 2.92/105,3.12/105 and 3.43/105 from 2016 to 2018 in Anyang,the incidence showed an increasing trend.Conclusions The GM (1,1) has high precision for predicting incidence of brucellosis.We need to take into account the epidemic characteristics of infectious diseases and social factors in actual practice.

Objective To investigate the surgical reparation of large coloboma raw surface after facial tumour resection in elderly patients.Methods According to the position and characteristics of tumor as well as the age and tolerance of the patients, full thick skin graft, the skin flaps with subcutaneous pedicle and free skin flap were designed and used in the reparation.Results 24 cases with large coloboma raw surface (5 cm×7 cm-12 cm× 16 cm)were treated by the utilization of three approaches after tumor resection.The large coloboma raw surface in all patients achieved the healing with satisfactory appearance.Conclusions After facial tumour resection, the large coloboma raw surface can be repaired by using the skin graft, skin flaps after tumor resection or free skin flap if designed reasonably.The procedure of operation is simple and the therapeutic effect is satisfactory.

Objective To investigate the radiosensitization effect of antihelminthic niclosamide on human triple-negative breast cancer MDA-MB-231 cells and the potential mechanism related to Wnt/β-catenin signaling pathway.Methods Four methyl thiazolyl tetrazolium(MTT) assay was used to measure the effect of niclosamide on cell viability at different concentrations and 50％ inhibitory concentration(IC50)value was calculated.MDA-MB-231 cells were divided into 4 groups:untreated control,niclosamide treatment alone group,radiation alone group and niclosamide plus radiation treatment group.The cells with or without 1.0 and 1.5 μmol/L niclosamide pre-treatment were irradiated with 137Cs γ-rays at doses of 0,2,4 and 6 Gy.Cell survival was assayed with the colony formation method,radiation-induced γH2AX foci was analyzed with immunofluorescence,cell cycle progression was assayed with flow cytometry,and the changes of phospho-and non-phospho-β-catenin and Cyclin D1 protein expressions were measured with Western blot.Results Niclosamde obviously inhibited the viability of MDA-MB-231 cells in a dosedependent manner with a IC50 value of 13.63 μmol/L.Pretreatment of cells with 1.0 and 1.5 μmol/L niclosamide evidently enhanced the radiosensitivity of MDA-MB-231 cells to γ-rays,and the values of SER were 1.37 and 1.62,respectively.Niclosamide pretreatment significantly increased radiation-induced γH2AX foci formation(t =3.91,P ＜0.05),diminished the radiation-induced G2/M arrest(t =8.05,P ＜0.01),and inhibited radiation-induced expressions of phospho-β-catenin (S675),non-phospho-β-catenin and Cyclin D1 proteins in MDA-MB-231 cells.Conclusions Niclosamide significantly can enhance the sensitivity of MDA-MB-231 cells to γ-ray irradiation through inhibiting Wnt/β-catenin signaling pathway,which results in the inhibition of DNA DSBs repair and the reduction of radiation-induced G2/M arrest.Wnt/β-catenin signaling pathway may serve as an ideal molecular target for radiosensitization of triplenegative breast cancer.

Objective To investigate the characteristics of repair of DNA double strand breaks (DSB) induced by high-LET α-particle irradiation and their relationship with chromatin structure in the G0 lymphocytes of human peripheral blood,in order to provide the experimental basis for the judgement and dose evaluation of internal α-particle radiation.Methods Peripheral whole blood were collected from four healthy adults and lymphocytes were separated.A monocellular layer of human lymphocytes attached in Mylar film were irradiated with 0 and 0.5 Gy of α-particles and the lymphocytes suspensions were irradiated with 0 and 0.5 Gy of γ-rays.The formations of γH2AX foci as a surrogate marker of DSB and Rad51 foci as a marker of homologous recombination (HR) repair and their spatial localization in chromatin structure were measured by immunofluorescence staining technique at 10 min-48 h post-irradiation.Results Linear-γH2AX foci tracks were observe at 10 min-2 h post-irradiation in lymphocytes exposed to α-particle irradiation(t =11.12,14.40,16.56,P ＜ 0.05),and almost completely disppeared at 6 h postirradiation.The frequencies of γH2AX foci peaked at 30 min after α-particle irradiation (t =51.72,P ＜0.05) and then decreased rapidly during 6 h post-irradiation (t =29.83,P ＜ 0.05).The average number of foci remained only about 16％ at 24-48 h post-irradiation.Moreover,27％ of γH2AX foci located at DAPI-bright heterochromatin region at 10 min after α-particle radiation,suggesting that the efficacy of DSB repair may be decreased.In contrast,at 10 min-48 h after γ-ray irradiation,no linear γH2AX foci track was observed and the γH2AX foci diffused randomly in nucleus and predominantly located in DAPI-weak euchromatin region.The numbers of formative and residual γH2AX foci after γ-ray irradiation were significantly less than those after α-particle radiation.During 30 min-2 h after α-particle and γ-ray irradiation,the frequencies of Rad51 foci slightly but not significantly increased in comparison with background level,and the frequencies of co-localization of Rad51 foci and γH2AX foci were only 3％-8％.Conclusions The formation of linear γH2AX foci tracks induced by high-LET α-particle irradiation in Go human lymphocyte could be used as biological indicator to estimate whether a person has been exposed to internal α-particle radiation.Prolonged persistence of residual γH2AX foci may be applicable for biological dosimetry.

Objective To evaluate the relationship between smoking and p53 gene mutation in the lung cancer patients with Meta-analysis.Methods PubMed,Web of Science,ProQest and Medline were used to search all the relevant studies about the association between smoking and p53 gene mutation in the patients with lung cancer. Based on reviewing full text,the studies were selected and evaluated and the data was extracted.Statistical analysis was performed with Stata 1 2.0 software including the heterogeneity inspection, publication bias assessment, sensitivity analysis,effect consolidating and cumulative Meta-analysis.Results Totally 15 articles with 1 770 lung cancer patients were identified. 69.6% of the patients were smokers,30.4% were non-smokers.Overall,the smokers with lung cancer had a 2.70-fold higher risk for p53 gene mutation than the non-smokers with lung cancer (OR=2.70,95%CI=2.04-3.59).Conclusion p53 gene mutation is associated with smoking in the patients with lung cancer.The smokers with lung cancer have higher risk for p53 mutation than non-smokers.

Objective To evaluate the association between BRAF600E mutation and pathological features in papillary thyroid carcinoma (PTC).Methods The raleted studies were searched through Medline,PubMed,and Web of Science,and meta-analysis was used to calculate the OR and 95％ CI of the study results.Results A total of 52 studies including 12 029 PTC patients was identified.The prevalence of BRAFV600EE mutation in PTC was 57.85％.There was a closed association between the BRAFV600E mutation and pathological features,including advanced TNM stage (OR =1.89,95％ CI 1.58-2.27),lymph node metastasis (OR =1.74,95％ CI 1.42-2.14),multifocality (OR =1.25,95％ CI 1.07-1.46),and recurrence (OR =2.26,95％ CI 1.25-4.09) of PTC.For Asians with PTC,the association between the BRAFV600E mutation and pathological features was indicated for advanced TNM stage(OR =1.56,95％ CI 1.24-1.96) and lymph node metastasis(OR =1.57,95％ CI 1.25-1.99).For Europeans with PTC,the association between the BRAFV600E mutation and pathological features was indicated for advanced TNM stage (OR =2.63,95％ CI 2.10-3.30),lymph node metastasis(OR =1.97,95％ CI 1.19-3.25),and multifocality (OR =1.35,95％ CI 1.01-1.80).Conclusion There were associations between the BRAFV600EE mutation and advanced TNM stage,lymph node metastasis,multifocality,and recurrence of PTC.