Objective To investigate the mutational spectrum and phenotype of abnormal hemoglobin in the diabetes screening population in Xi'an Region. Methods A total of 30356 patients from the Second Affiliated Hospital of Xi'an Jiaotong University were collected and measured for HbA1c by high-performance liquid chromatography (HPLC) from January 2022 to December 2022. Both complete blood count and hemoglobin analysis were carried out when HPLC suggested an abnormal peak. Abnormal hemoglobins were identified by Sanger sequencing. Results In this study,the rate of abnormal hemoglobin in the diabetes screening population of Xi'an region was 0.096％ (28/30356). A total of 7 types of abnormal hemoglobins were detected:Hb E (n=10),G-Coushatta (n=6),Hb G-Taibei(n=6),Hb Q-Thailand (n=2),Hb North Manchester (n=2),Hb Ottawa (n=1) and Hb Queens (n=1). One case was Hb G-Taipei combined with IVS Ⅱ-659_664 (-GCAATA),the first report in the world. The abnormal hemoglobin phenotypes were normal except for seven cases of Hb E. Except for Hb G-Coushatta and Hb Ottawa,which did not interfere with the glycation test,the other five abnormal hemoglobins affected the glycation test results. Conclusion There is a certain prevalence rate of abnormal hemoglobin in the diabetes screaning population of Xi'an,mainly Hb E,Hb G-Taibei and Hb G-Coushatta. Most abnormal hemoglobins present normal phenotypes,but they could interfere with glycation test results.

Objective To observe the effect of self-formulated Tongluo Yizhi formula on memory deficits and endoplasmic reticulum stress (ERS) in rats with vascular dementia (VD) and its mechanism. Methods Fifty SPF-grade Wistar rats were randomly divided into sham surgery group,model group,TLYZF low-dose (12.5 mg/kg/d) group,TLYZF medium dose (25 mg/kg/d) group and TLYZF high-dose (50 mg/kg/d) group,with 10 rats in each group. Except for the Sham group,other rats were subjected to modified bilateral carotid artery ligation to construct VD models. After 4 weeks of TLYZF intervention,Morris water maze experiment was used to evaluate the learning and memory abilities of rats.Hematoxylin eosin staining was used to detect the pathological structure of the hippocampus.Nissl staining was used to detect hippocampal neuronal damage.In situ end transfer enzyme labeling staining was used to detect hippocampal neuronal apoptosis.Enzyme-linked immunosorbent assay was used to detect the content of inflammatory factors in hippocampal tissue.Protein immunoblotting was used to detect apoptosis of hippocampal tissue cells and the expression of ERS related proteins.Results Compared with the Sham group,the average escape latency of the model group was prolonged(t=14.059),the number of crossing the platform was reduced(t=8.534),the hippocampal neurons were disorderly arranged,the morphology was fuzzy,the cytoplasmic vacuolization,the nuclear pyknosis,the inflammatory cell infiltration increased,the number of Nissl bodies decreased(t=17.131),and the neuronal apoptosis rate increased(t=17.701). The contents of tumor necrosis factor-α (TNF-α),interleukin-1 β (IL-1β) and IL-6 increased(t=6.541,6.957,10.014),the expression of Bcl-2 associated X protein (Bax),cleaved caspase-3,glucose-regulated protein 78 (GRP78),phosphorylated endoplasmic reticulum kinase (p-PERK)/PERK,phosphorylated eukaryotic initiation factor 2α (p-eIF2α)/eIF2α,activating transcription factor 4 (ATF4) and CCAAT/enhancer binding protein (C/EBP) homologous protein (CHOP) increased(t=13.548～76.468),while the expression of B-cell lymphoma-2 (Bcl-2) proteinwas decreased (t=39.691),and the difference were statistically significant (all P<0.05),respectively. Compared with the model group,the average escape latency of TLYZF-treated rats was shortened(t=2.476,7.266,11.306),the number of platform crossing was increased(t=2.187,4.471,6.932),the hippocampal neurons were relatively regular,the nucleus was clear,the inflammatory cell infiltration was significantly reduced,the content of Nissl bodies was increased(t=4.359,9.477,11.449),the apoptosis rate of neurons was decreased(t=3.631,6.145,7.580),the contents of TNF-α,IL-1β and IL-6 were decreased (t=2.382～8.293),the expression of Bax(t=4.696,16.250,20.250),cleaved caspase-3(t=15.205,27.000,26.833),GRP78(t=5.918,13.139,13.741),p-PERK/PERK(t=13.416,14.230,17.889),p-eIF2α/eIF2α(t=20.152,39.346,50.750),ATF4(t=12.093,24.395,22.946)and CHOP protein((t=21.592,19.207,22.136)decreased,while Bcl-2 protein increased(t=7.474,20.761,35.350),the differences were statistically significant (all P<0.05),respectively,and the high-dose group had the best therapeutic effect.Conclusion Self-formulated Tongluo Yizhi formula could reduce the apoptosis and inflammatory injury of hippocampal neurons in VD rats,and its mechanism may be related to inhibiting ERS mediated by PERK.

Objective To explore the effect and mechanism of long non-coding RNA (LncRNA) nuclear enriched abundant transcript 1 (NEAT1) on proliferation and metastasis of myeloma through miR-195-5p/CCAAT enhancer binding protein α (CEBPA) axis. Methods ① From March 2021 to February 2023,bone marrow mononuclear cells (BMNC) from 40 patients with multiple myeloma (MM) and 10 healthy bone marrow donors who were hospitalized in the Department of Hematology the Second People's Hospital of Liaocheng City,Shandong First Medical University and MM cell lines U266,RPMI 8226,NCI-H929 and MM.1S were collected. RT-qPCR and Western blot were used to detect NEAT1,miR-195-5p,CEBPA mRNA and protein levels in cells. ② U266 cells were divided into overexpressing NEAT1 group (NEAT1 group) and its control group (NC group),knockdown NEAT1 expression group (sh-NEAT1 group) and its control group (sh-NC group),overexpressing NEAT1 and miR-195-5p group (NEAT1+miR-195-5p group) and its control group (NEAT1+miR-NC group),and overexpressing miR-195-5p and CEBPA group (miR-195-5p+CEBPA group) and its control group (miR-195-5p+NC group). CCK-8 and EdU staining were used to detect cell proliferation ability. Transwell assay was used to detect cell migration and invasion ability. RT-qPCR was used to detect NEAT1,miR-195-5p,and CEBPA mRNA levels in cells. Western blot was used to detect CEBPA,Ki67,proliferating cell nuclear antigen (PCNA),matrix metalloproteinase (MMP)-2,MMP-9,phosphoinositide 3 kinase (PI3K),p-PI3K,protein kinase B (AKT),p-AKT,mechanistic target of rapamycin (mTOR),and p-mTOR protein levels. ③ Twenty nude mice were divided into knockdown NEAT1 expression group (sh-NEAT1 group) and its control group (sh-NC group),with ten mice in each group. Nude mice were subcutaneously injected with corresponding transfected U266 cell suspension,and the differences in various indicators of transplanted tumor were measured after 4 weeks. Results ① Compared with normal BMNC,NEAT1,CEBPA mRNA and protein levels in MM patients and MM cell lines were increased,while miR-195-5p level was decreased,and the differences were significant (t=11.697,9.272,4.352,11.639,all P<0.05). ② After overexpression of NEAT1,NEAT1,CEBPA mRNA and protein levels in cells were increased (t=12.825,5.874,13.893),while miR-195-5p level was decreased (t=4.797),the A value and EdU positive rate after 72 hours of cell culture were increased (t=9.425,5.632),the number of migration/invasion cells were increased (t=8.841,5.364),and Ki67,PCNA,MMP-2,MMP-9,p-PI3K/PI3K,p-AKT/AKT and p-mTOR/mTOR protein levels were increased (t=14.227,7.743,7.348,7.803,8.714,8.629,7.359),with significant differences (all P<0.05). After knockdown the expression of NEAT1,NEAT1,CEBPA mRNA and protein levels in cells were decreased (t=5.776,5.001,4.503),while miR-195-5p level was increased (t=4.456),the ability of cell proliferation,invasion and metastasis was decreased (t=6.204,8.792),with significant differences (all P<0.05). Overexpression of miR-195-5p could partially reverse the effects of overexpression of NEAT1 on the aforementioned cellular indicators,and the differences were significant (t=4.356～10.809,all P<0.05). Meanwhile,overexpression of CEBPA could partially reverse the effects of overexpression of miR-195-5p on the aforementioned cellular indicators,and the differences were significant (t=4.329～14.452,all P<0.05). ③ Knockdown NEAT1 expression could inhibit the growth of transplanted tumors in nude mice,and the differences were significant (t=5.175～18.190,all P<0.05). Conclusion LncRNA NEAT1 promotes proliferation,invasion and metastasis of MM cells by targeting miR-195-5p/CEBPA expression,which may act by activating the PI3K/AKT/mTOR pathway.

Objective To analyze the relationship between 15 immunophenotypes of peripheral blood T cells and age and gender in healthy adults in the Dalian area. Methods A total of 277 healthy adults admitted to the physical examination center of the Second Affiliated Hospital of Dalian Medical University from October 2022 to June 2023 were selected as the research subjects,including 154 males and 123 females. They were divided into three groups according to age:young group(18～44 years,n=103),middle-aged group(45～60 years,n=114) and old age group(>60 years,n=60). Flow cytometry was used to determine immunophenotypes of T cells,including the absolute count and proportion of na?ve cells (N),central memory (CM),effector memory (EM),terminal effector memory (TEM),activation (HLA-DR+) and senescence (CD28-) CD4+and CD8+T cells,and the differences of them among age and gender were analyzed. Spearman correlation analysis was also used to evaluate the correlation between age and immunophenotypes of T cells. Results Compared with the young group,the absolute count and proportion of CD8+TEM,CD4+HLA-DR+,CD8+HLA-DR+and CD8+CD28-T lymphoaytes cells were increased in the middle-aged group and old age group (Z=2.009～6.607),while the absolute count and proportion of CD8+N T cells were decreased in the middle-aged group (Z=5.574～7.999) and old age group,and the differences were statistically significant (all P<0.05),respectively. Compared with the female group,the absolute count and proportion of CD8+CM,CD8+HLA-DR+and CD8+CD28-T cells were increased in the female group (Z=2.945～6.131),while the absolute count and proportion of CD3+,CD4+and CD8+N T cells were decreased in the male group (Z=2.075～4.225),and the differences were statistically significant(all P<0.05),respectively. Spearman correlation analysis showed that age was positively correlated with the absolute count and proportion of CD4+CM,CD4+HLA-DR+,CD4+CD28-,CD8+TEM,CD8+HLA-DR+and CD8+CD28-T lymphocytes cells (r=0.125～0.479,all P<0.05),while negatively correlated the absolute count and proportion of CD4+N and CD8+N T lymphocytes cells (r=-0.538～-0.148,all P<0.05). Conclusion The 15 immunophenotypes of peripheral blood T lymphocytes cells in healthy adults from Dalian area are affected by age and gender,so it is necessary to establish a suitable local reference interval to provide a more accurate reference for immune function assessment.

Objective To understand the membrane protein molecular epidemiology of carbapenem-resistant Pseudomonas aeruginosa (CRPA) in the region,and provide some evidence for rational drug use or application of efflux pump inhibitors. Methods Collected Pseudomonas aeruginosa isolated from four hospitals in the region from October 2022 to August 2023,and used SYBR-PCR method to quantitatively detect the relative mRNA expression (RE) levels of 10 membrane protein coding genes,including mexA,B,C,D,E,F,X,Y,and oprD,M. Then categorized the strains into five groups based on ceftazidime,cefepime,imipenem,and meropenem resistance phenotype combination,including the compassionate group (Group Ⅰ),Group Ⅱ with full resistance,IPM,MEM resistant,CAZ and CFP sensitive groups (Group Ⅲ),IPM resistance,MEM non-resistance (sensitive or intermediate) group (Group Ⅳ),IPM,MEM resistance,CAZ and CFP non-resistance groups (Group V).The median RE of each membrane protein-coding gene was analyzed. Results A total of 108 strains of Pseudomonas aeruginosa were collected,with 24 strains in Group Ⅰ as controls and 84 strains in the carbapenem resistant group,including 32 strains in Group Ⅱ,22 strains in Group Ⅲ,13 strains in Group Ⅳ,and 17 strains in Group Ⅴ. The expression of mexD,mexE,mexF,mexX and mexY in the drug-resistant group was higher than that in the control group,and the differences were statistically significant (U=409.5～661.0,all P<0.05). There was no statistically significant difference in mexA,mexB,mexC,oprD and oprM with the control group (U=767.0～1004.5,all P>0.05). There was no significant difference in the expression of RE genes encoding various membrane proteins among strains from different hospitals (H=0.914～7.407,all P>0.05). Among the four different phenotypes,there was no statistically significant difference in the irregular distribution of mexA and oprM RE between each group and the control group (UmexA=95.0～264.0,UoprM=143.0～331.0). The mexC RE in each group was lower than that in the control group,but the differences were not statistically significant (U=134.0～344.5,all P>0.05). MeixE and meixY RE were both higher than the control group,and the differences were statistically significant (UmexE=48.0～230.0,UmexY=83.0～184.0). MeixB was lower than the control group in group Ⅳ (U=72.0),and the differences were statistically significant (all P<0.05). MeixD and meixF showed consistent expression,with higher expression in groups Ⅲ,Ⅳ and Ⅴ compared to the control group (UmeixD=34.0～102.0,UmeixF=65.0～113.0). MeixX was expressed higher in groups Ⅱ,Ⅳ and Ⅴ compared to the control group (U=164.0,58.0,111.0),while oprD was only expressed lower in group Ⅲ than in the control group (U=140.0),with statistically significant differences (all P<0.05). Although the expression of oprD in groups Ⅱ,Ⅳ and Ⅴ was lower than that in the control group,the differences were not statistically significant (U=381.0,102.0,144.0,all P>0.05). Conclusion ExCD,mexEF and mexXY are the main membrane protein combinations of CRPA efflux pumps in Kunming area. Upregulation of mexD,E,F,X,and Y membrane protein expression enhanced efflux. The correlation between mexAB oprM efflux pump and carbapenem resistance in CRPA in this area was low. The low expression of oprD played a role in the efflux mechanism in strains that do not produce β-lactase,but there was no significant difference in low expression in enzyme producing strains.

Objective To investigate the role of long non-coding RNA 01004 (LncRNA01004) in accelerating the malignant progression of breast cancer cells and its potential regulatory mechanism. Methods Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the expression level of LncRNA01004 in breast cancer tissues and cells. The Starbase online database predicted the binding of LncRNA01004 to CPSF1 and verified it through RNA binding protein immunoprecipitation(RIP) analysis. MCF-7 cells were transfected with LncRNA01004 interference sequence (sh-LncRNA01004) or overexpressed vector (LncRNA01004),or co-transfected with Cleavage and Polyadenylation Specific Factor 1 (CPSF1) interference sequence (sh-CPSF1). Cell viability,invasion and apoptosis were detected with CCK-8,Transwell and flow cytometry (FCM). The overexpression and silencing efficiency of LINC01004 and CPSF1 were detected by qPCR. Western blot (WB) analysis of CPSF1 protein,apoptosis-related protein[B cell lymphoma/leukemia-2(Bcl-2),Bcl-2 Associated X(Bax)]and Epithelial-Mesenchymal transition (EMT) related protein[Epitheia-cadherin(E-cadherin),Nerve-cadherin(N-cadherin),Vimentin,zinc-finger transcription factor(Snail)],and the activity of Cysteinyl aspartate-specific proteinase-3 (Caspase-3) was determined by enzyme-linked immunosorbent assay. Results LncRNA01004 was significantly up-regulated in breast cancer tissues (5.14±0.33) compared with paracancer tissues (1.02±0.03),with the statistically significant difference (t=-78.637,P<0.001);LncRNA01004 expression in breast cancer cells was significantly higher than that in normal breast epithelial cells,the difference between groups is statistically significant (F=142.248,P<0.001). Compared with the Control group,LncRNA01004 significantly inhibited the proliferation (42.15±2.11 vs 100.02±0.65) and invasion (18.65％±1.44％ vs 41.36％±1.57％) of MCF-7 cells,induced apoptosis (16.58％±1.52％ vs 5.24％±1.12％),increased the activity of Caspase-3 (2.93±0.711. vs 51±0.43) and the expression of Bax (2.74±0.39 vs 1.01±0.02) protein,and inhibited the expression of BcL-2 (0.32±0.07 vs 1.02±0.03) protein,with the statistically significant difference (t=3.075～19.332,all P<0.05). Significantly increased compared with Control group,the silent LncRNA01004 E-cadherin (3.06±0.37 vs 1.01±0.02) protein levels,lower N-cadherin (0.44±0.11 vs 1.00±0.01),Vimentin (0.39±0.13 vs 1.02±0.03) and Snail(0.30±0.08 vs 1.01±0.03) protein levels,with the statistically significant difference (t=9.989～17.164,all P<0.05).LncRNA01004 binds to CPSF1 and promotes CPSF1 protein expression.Silencing CPSF1 inhibited the proliferation and invasion of MCF-7 cells,induced cell apoptosis,and counteracted the effect of LncRNA01004 overexpression on MCF-7 cells. Conclusion LncRNA01004 may promote EMT through up-regulation of CPSF1,and then promote proliferation and invasion of breast cancer cells,inhibit cell apoptosis,and participate in the malignant progression of breast cancer.

Objective To analyze cysteinylaspartate specific proteinase-3,caspase-7,caspase-9,AMP activated protein kinase(AMPK)in the diagnosis and prognosis evaluation of sepsis.Methods Sixty patients with sepsis from the Affiliated Hospital of Xiangnan University from March 2020 to March 2023 were selected as the observation group,and another 60 healthy individuals from the physical examination center of the hospital during the same period were selected as the control group.Acute physiology and chronic health evaluationⅡ(APACHEII)score was used to evaluate the severity of the patient's condition.Serum caspase-3,caspase-7,caspase-9,AMPK and APACHEII scores were detected,and compared between the two groups.These parameters were compared with serum Caspase-3,caspase-7,caspase-9,AMPK and APACHEII scores in different disease severity groups.The patients were divided into survival group(50 cases)and death group(10 cases)according to prognosis.Serum Caspase-3,caspase-7,caspase-9,AMPK and APACHEII scores were compared between the two groups,and the correlation between serum caspase-3,caspase-7,caspase-9,AMPK and APACHEII scores were analyzed by Pearson.Receiver operating characteristic(ROC)was drawn,the area under the curve(AUC)was calculated,the predictive efficacy of serum caspase-3,caspase-7,caspase-9 and AMPK in patients death with sepsis were analyzed.Results The serum levels of caspase-3,caspase-7,caspase-9,AMPK,and APACHEII score in the observation group were higher than those in the control group,and the differences were statistically significant(t=19.501～44.219,all P<0.05).The serum caspase-3,caspase-7,caspase-9,AMPK,and APACHEII scores in the sep-tic shock group were higher than those in the severe and mild groups,and the differences were statistically significant(t=2.091～6.946,all P<0.05).The serum caspase-3,caspase-7,caspase-9,AMPK,and APACHEII scores of the death group were higher than those of the survival group,and the differences were statistically significant(t=4.576～8.835,all P<0.05).Serum caspase-3,caspase-7,caspase-9 and AMPK were positively correlated with APACHEII scores(r=0.382～0.406,all P<0.05).The AUC of combined detection of serum caspase-3,caspase-7,caspase-9,and AMPK for predicting death in sepsis patients was 0.863,with 95%confidence interval of 0.805-0.958.which was greater than that of single indicator detection,and the difference is statistical-ly significant(Z=2.912～2.294,all P<0.05).Conclusion Serum caspase-3,caspase-7,caspase-9,and AMPK in sepsis patients showed abnormally high expressions,which were closely related to disease severity and prognosis.Combined detection could improve the sensitivity and specificity of death prediction,which has certain reference value.

Objective To explore the predictive value of systemic immune-inflammation index(SII)and control of nutritional status(CONUT)score for the efficacy of immune combined chemotherapy in patients with advanced or recurrent gastric cancer.Methods A retrospective analysis was conducted on the clinical data of 143 patients with advanced or recurrent gastric cancer who underwent immune combined chemotherapy as a treatment regimen at Huangshan People's Hospital from January 2020 to December 2023.The patients were divided into a disease control group(n=105)and a disease non-control group(n=38)according to the response evaluation criteria in solid tumor(RECIST).The CONUT score and SII levels between the two groups were compared.Multivariate Logistic regression analysis was used to identify the factors influencing the efficacy of immune combined chemotherapy in patients with advanced or recurrent gastric cancer.Receiver operating characteristic(ROC)curves were employed to predict the value of pre-treatment CONUT score,SII,and their combination in predicting the efficacy of immune-combined chemotherapy for advanced or recurrent gastric cancer.Results The CONUT score(2.8±0.5)and SII level(1 073.64±141.26)in the disease control group under than those in the disease non-control group(3.3±0.7,1 183.27±186.34),and the differences were statistically significant(t=4.721,3.751,all P<0.05).Multivariate Logistic regression analysis revealed that age,degree of differentiation,maximum diameter of measurable lesions,CONUT score,and SII were all factors influencing the treatment outcome in patients with advanced or recurrent gastric cancer(Wald χ2=5.161～22.758,all P<0.05).The ROC curve results showed that the AUC(95%CI)for pre-treatment CONUT score,SII,and their combination in predicting the efficacy of immuno chemotherapy in patients with advanced or recurrent gastric cancer were 0.746(0.667～0.815),0.839(0.768～0.895)and 0.875(0.809～0.924),respectively,with sensitivities of 73.11%,76.32%and 79.68%,and specificities of 91.43%,82.86%and 84.75%,respectively.Conclusion CONUT score and SII are influencing factors for the efficacy of immuno-combined chemotherapy in patients with advanced or recurrent gastric cancer,and their combination demonstrates good predictive value for the treatment outcome.

Objective To investigate the relationship between the expression of discoidin,CUB and LCCL domain containing protein 1(DCBLD1)and cytoskeleton associated protein 2(CKAP2)in cervical cancer(CC)tissues and epithelial mesenchymal transition(EMT)and clinical prognosis.Methods 94 CC patients diagnosed and treated in Shiyan Hospital of Traditional Chinese Medicine from February 2017 to February 2019 were selected.The expression of DCBLD1 messenger ribonucleic acid(mRNA),CKAP2 mRNA,N-cadherin(N-cad)mRNA,vimentin(Vim)mRNA and TWIST mRNA in tissues was detected by real-time fluorescence quantitative PCR(qRT-PCR).The expressions of DCBLD1 and CKAP2 were detected by immunohistochemistry(IHC).Pearson correlation analysis was used to analyze the relationship between DCBLD1 mRNA,CKAP2 mRNA and EMT-related indicators.Kaplan-Meier survival curve was drawn to compare the prognosis of CC patients with different DCBLD1 mRNA and CKAP2 mRNA expression.COX regression analysis was used to analyze the prognostic factors of CC patients.Results The expression of DCBLD1 mRNA,CKAP2 mRNA,N-cad mRNA,Vim mRNA and TWIST mRNA in CC cancer tissues was higher than that in adjacent tissues,and the differences were statistically significant(t=32.763～52.824,all P<0.05).The positive rates of DCBLD1 protein(89.36%)and CKAP2 protein(87.23%)in CC cancer tissues were higher than those in adjacent normal tissues(7.45%,6.38%),and the differences were statistically significant(χ2=126.278,123.396,all P<0.001).The expression of DCBLD1 mRNA and CKAP2 mRNA in CC cancer tissues were positively correlated with the expression of N-cad mRNA,Vim mRNA and TWIST mRNA(r=0.655～0.744,all P<0.001).The expression of DCBLD1 mRNA and CKAP2 mRNA in patients with FIGO stage IB2～IIB and lymph node metastasis were higher than those in patients with stage IA～IB1 and without lymph node metastasis(t=25.644～35.674,all P<0.05).The 5-year progression free survival rates of the high expression groups of DCBLD1 mRNA and CKAP2 mRNA were 63.04%and 62.22%,respectively,which were lower than those of the low expression groups of DCBLD1 mRNA and CKAP2 mRNA(91.67%and 91.84%),respectively,and the differences were statistically significant(Log-Rank χ2=7.181,6.527,all P<0.05).FIGO stage IB2～IIB,high DCBLD1 mRNA and high CKAP2 mRNA were risk factors affecting the prognosis of CC patients(Wald χ2=8.277,15.877,10.927,all P<0.05).Conclusion The expression of DCBLD1 and CKAP2 in CC cancer tissues is significantly increased,which is related to EMT related indicators and plays a promoting role in the progression of CC tumors.They are new prognostic markers for CC.

Objective To screen the differentially expressed circular RNA(circRNA)in peripheral blood of patients with type 2 diabetic nephropathy(T2DN)and study the correlation between the expression of circAKT3 level and renal fibrosis.Methods A retrospectively study was conducted on 176 T2DM patients admitted to Mianyang 404 Hospital from December 2022 to December 2023 based on whether the patients developed DN,they were divided into a simple T2DM group(n=60)and DN group(n=116).At the same time,30 healthy subjects who underwent physical examination in Mianyang 404 Hospital during the same period were selected as the control group.Clinical data of all subjects were collected.DN patients were divided into microalbuminuria group(M-DN group,n=66)and macroalbuminuria group(MS-DN group,n=50)according to Mogensen classification criteria.Twenty samples were randomly selected from the control group,M-DN group,T2DM group and MS-DN group,and 2 ml peripheral venous blood was extracted for RNA extraction and circRNA microarray scanning analysis to screen the differentially expressed circRNA.The correlation between the most significant differential circRNA expression level and renal fibrosis markers and renal function indicators was statistically analyzed.Receiver operator characteristic(ROC)curve was used to analyze the diagnostic efficacy of circAKT3 for DN.Results Compared with the control group,the level of circAKT3 in MS-DN patients was down-regulated by 9.72 times,which was the most significantly down-regulated circRNA.qRT-PCR showed that compared with the control group,the relative expression of circAKT3 in the T2DM group,M-DN group and MS-DN group was 0.85±0.23,0.54±0.13 and 0.12±0.09,respectively,with the progression of DN,the relative expression level of circAKT3 in peripheral blood of patients gradually decreased,and the differences were statistically significant(t=5.100,8.568,10.960,all P＜0.001).Spearman correlation test showed that the level of circAKT3 in peripheral blood of DN patients was correlated with the duration of diabetes,serum Creatinine,BUN,cystatin C,TGF-β,CIV,PCIII and Alb were negatively,correlated(r=-0.756～-0.621,all P＜0.05).Glomerular filtration rate was positively correlated(r=0.695,P=0.010).ROC curve analysis showed that the sensitivity and specificity of circAKT3 in the diagnosis of DM were 75.3%and 71.1%,respectively,and the AUC was 0.792.The sensitivity,specificity and AUC for M-DN were 90.3%,85.7%and 0.875,respectively,and the sensitivity,specificity and the AUC for MS-DN were 88.2%,87.6%and 0.916,respectively.Conclusion The level of serum circAKT3 is significantly down-regulated in DN patients,which can be used as a potential auxiliary diagnostic marker of DN.