Objective To investigate the expression levels of serum exosomal LncRNA GAS5 in gastric cancer(GC)patients and eval-uate its clinical significance.Methods Seventy GC patients admitted to Henan Electric Power Hospital from January 2019 to Decem-ber 2020 and 70 age-and sex-matched healthy controls undergoing physical examination during the same period were enrolled.Their se-rum samples and clinicopathological data were collected.Additionally,cancer tissue and adjacent normal tissue specimens were ob-tained from 20 GC patients who underwent surgical resection.The expression levels of LncRNA GAS5 in tissues and serum exosomes were detected by real-time fluorescence quantitative PCR(qRT-PCR).The chemiluminescence method was used to detect the levels of serum carcinoembryonic antigen(CEA)and carbohydrate antigen 19-9(CA19-9).The receiver operating characteristic(ROC)curve was used to assess the diagnostic efficacy of different detection indicators for GC.The correlations of serum exosomal LncRNA GAS5 levels with clinicopathological parameters and prognosis of GC patients were also analyzed.Results The expression levels of serum ex-osomal LncRNA GAS5 in GC patients were significantly lower than that in healthy controls(t=10.84,P＜0.001).The expression levels of LncRNA GAS5 in GC tissues were also significantly lower than that in adjacent tissues(t=14.94,P＜0.001).The analysis results of the ROC curve showed that the area under the ROC curve(AUCROC)of serum exosomal LncRNA GAS5 for the screening of GC was 0.807.When the cut-off value was 1.96,its sensitivity was 81.43%and specificity was 67.14%.The diagnostic efficacy of serum exoso-mal LncRNA GAS5 for GC was significantly better than traditional tumor markers CEA and CA19-9,and their AUCROC were 0.664 and 0.713,respectively.The analysis results of clinicopathological parameters of GC patients showed that low serum exosomal LncRNA GAS5 levels were significantly correlated with large tumor size,advanced TNM stage,and distant metastasis(P＜0.05).The overall survival time of patients with low expression of serum exosomal LncRNA GAS5 was significantly lower than that of patients with high ex-pression(median overall survival time:23 vs 35 months,P=0.03).Conclusion Serum exosomal LncRNA GAS5 in GC patients is down-regulated,which has good diagnostic efficacy for GC and is expected to become a potential biological marker for the diagnosis and prognosis evaluation of GC patients.

Objective To investigate the diagnostic value of serum carcinoembryonic antigen-related cell adhesion molecule 6(CEACAM6)in pancreatic cancer.Methods A total of 188 patients admitted to the Pancreas Center of the First Affiliated Hospital of Nanjing Medical University from May to December 2024 were consecutively enrolled,including 112 patients in the pancreatic cancer group and 76 patients in the benign pancreatic disease group(43 with pancreatitis and 33 with benign pancreatic tumors).Additional-ly,76 healthy individuals were included in the health control group.Serum CEACAM6 and carbohydrate antigen 19-9(CA19-9)levels were detected by ELISA and chemiluminescence immunoassay(CLIA),respectively.The receiver operating characteristic(ROC)curve was drawn to evaluate the diagnostic efficacy of serum CEACAM6 for pancreatic cancer.The positive rate of serum CEACAM6 in CA19-9-negative pancreatic cancer patients was also investigated.Results The levels of serum CEACAM6 in the pancreatic cancer group(20.3[11.1,34.0]ng/mL)were significantly higher than those in the benign pancreatic disease group(6.2[2.6,10.5]ng/mL,Z=64.1,P＜0.001)and healthy control group(0.8[0.4,4.4]ng/mL,Z=131.6,P＜0.001).The areas under the ROC curve(AUCROC),sensitivity,and specificity of CEACAM6 and CA19-9 in the diagnosis of pancreatic cancer were 0.889(95%CI:0.854-0.924)and 0.819(95%CI:0.761-0.863),81.3%and 65.2%,and 84.9%and 86.2%,respectively.The AUCROC,sensitivity,and specificity of the combined detection of the two reached 0.916(95%CI:0.875-0.946),91.1%,and 82.2%,respectively,which were better than any single indicator(P＜0.05).In addition,the positive rate of serum CEACAM6 in CA19-9-negative pancreatic canc-er patients was 83.3%,significantly higher than CEA and CA125(P＜0.01).Conclusion The detection of serum CEACAM6 is help-ful to the diagnosis of pancreatic cancer,and its combination with serum CA19-9 can improve the diagnostic efficacy of pancreatic cancer.

Objective To investigate the expression levels of serum hemoglobin β(HBB)in hepatocellular carcinoma(HCC)patients and its correlation with topoisomeraseⅡα(TOP2A)gene.Methods A total of 48 HCC patients visited the Second Affiliated Hospital of Nantong University from August 2023 to September 2024 were selected as the HCC group,and 32 healthy individuals who underwent physical examination during the same period were selected as the healthy control group.Their blood samples were collected,and the ex-pression levels of serum HBB and TOP2A genes were detected by real-time fluorescence quantitative polymerase chain reaction(qRT-PCR).Then,the relationship between the expression of HBB gene and clinicopathological parameters of the patients was ana-lyzed.The Kaplan-Meier Plotter database was used to evaluate the relationship between the expression of HBB gene and the prognosis of HCC.The diagnostic value of the expressions of serum HBB and TOP2A genes for HCC was assessed by the receiver operating charac-teristic(ROC)curve.Spearman rank correlation analysis was used to evaluate the correlation between the expressions of serum HBB and TOP2A genes in HCC patients.The regulatory effect of HBB gene on TOP2A gene was verified by the cell experiment.Results The expression levels of serum HBB gene in HCC patients(0.097[0.055,0.155])were significantly lower than that in healthy controls(1.029[0.625,1.434],U=19,P＜0.001).The expression levels of serum TOP2A gene in HCC patients(1.810[0.825,3.623])were significantly higher than that in healthy controls(1.047[0.604,1.364],U=495,P=0.007).The expression level of serum HBB gene in HCC patients was significantly negatively correlated with that of TOP2A gene(ρ=-0.384,P=0.007).The analysis results of clinicopathological parameters showed that the expression level of HBB gene was only related to tumor size(χ2=4.090,P＜0.05).The Kaplan-Meier survival analysis showed that the 5-year overall survival rate of the patients with low expression of HBB gene was signifi-cantly lower than that with high expression(HR=0.680,95%CI:0.470-0.970,P＜0.05).The analysis of the ROC curve showed that the area under the ROC curve(AUCROC)of HBB gene in diagnosing HCC was 0.987(95%CI:0.963-1.000).When the cut-off value was 0.228,its sensitivity was 100%and specificity was 97%.The AUCROC of TOP2A gene for the diagnosis of HCC was 0.677(95%CI:0.559-0.797).When the cut-off value was 1.285,its sensitivity was 65%and specificity was 75%.The combined detection of HBB and TOP2A genes for the diagnosis of HCC had an AUCROC of 0.988(95%CI:0.965-1.000).When the cut-off value was 0.657,its sensitivity was 100%and specificity was 97%.The cell experiment results showed that the overexpression of HBB gene could inhibit the expression of TOP2A gene,while the knockout of HBB gene had the opposite effect.Conclusion HBB gene is lowly expressed in the serum of HCC patients and is significantly negatively correlated with the expression of TOP2A gene.

Objective The causal relationship between immune cells and hepatocellular carcinoma(HCC)risk was investigated using a two-sample Mendelian randomization method.Methods The datasets including 731 immune cells and HCC were obtained from the GWAS database and Finngen database,respectively.The stability and reliability of Mendelian randomization studies were evaluated by the MR-Egger regression,MR PRESSO,Cochran's Q test,and leave one out test.The inverse variance weighting,MR-Egger,weigh-ted median,simple mode and weighted mode were used to investigate the causal relationship between immune cells and HCC.Results A total of 4 immune cells were found to have a potential causal relationship with HCC,and the results were stable.The CD3+CD39+Treg,CD80+granulocyte and CD4+Treg were protective factors for HCC(OR=0.910,95%CI:0.852-0.972;OR=0.919,95%CI:0.865-0.975;OR=0.924,95%CI:0.873-0.978),while CD45+CD33+HLA-DR+CD14+marrow cells were a risk factor for HCC(OR=1.116,95%CI:1.033-1.204).Conclusion The CD3+CD39+Treg,CD80+granulocytes,and CD4+Treg are negatively associated with the risk of HCC,while CD45+CD33+HLA-DR+CD14+marrow cells are positively associated with the risk of HCC.

Objective To investigate the distribution of serum specific IgG antibodies against food in children with autism spectrum disorders(ASD),and provide experimental evidence for improving gastrointestinal symptoms of ASD children.Methods A retrospec-tive study was conducted on 411 children with ASD visited the Department of Psychology and Behavior of Hebei Children's Hospital from January 2021 to December 2022.Additionally,631 healthy children who underwent physical examination during the same period were collected as the healthy control group.Their venous blood samples were collected,and ELISA was used to detect the levels of 14 kinds of serum specific IgG antibodies against food.The distribution characteristics of food intolerance in children with different genders and ages were investigated.Results The total positive rate of specific IgG antibodies against food in children with ASD reached 91.84%(405/441).The top 3 positive rates of specific IgG antibodies were against egg(71.89%),milk(56.70%),and wheat(56.02%),respectively,which were significantly higher than those in the healthy control group(χ2=42.81,27.48,and 26.88,re-spectively,P＜0.05).The comparative results of gender composition showed that the positive rate of specific IgG antibody against rice in female ASD children(15.96%)was significantly higher than that in male ASD children(4.90%,χ2=11.84,P＜0.05).The posi-tive rates of specific IgG antibodies against egg and wheat in male and female ASD children were significantly higher than those in the healthy control group(χ2=19.44 and 4.42 for males,χ2=4.66 and 10.93 for females,P＜0.05).The positive rate of specific IgG anti-body against milk in male ASD children was significantly higher than that in the healthy control group(χ2=12.62,P＜0.05),while those against soybean and rice in female ASD children were also significantly higher than that in the healthy control group(χ2=7.00 and 5.42,P＜0.05).There were significant differences in the positive rates of food intolerance to milk and soybean in ASD children with different ages(χ2=13.74 and 9.70,P＜0.05)and they decreased with age.The positive rates of specific IgG antibody against wheat in ASD children aged 2-3 and 4-6 years were significantly higher than those in the healthy control group(χ2=5.78 and 4.55,P＜0.05).The positive rate of specific IgG antibody against milk in ASD children aged 4-6 years was significantly higher than that in the healthy control group(χ2=7.57,P＜0.05).Conclusion The highest positive rate of specific IgG antibodies against food in ASD chil-dren is against egg,which is not related to the patient's gender.Next are milk and wheat.The gastrointestinal issues and related food intolerance should be taken into account in the management of patients with ASD.

Objective To establish the reference range for the chemiluminescence detection of N-terminal pro brain-type natriuretic peptide(NT-proBNP)in apparently healthy adults of Nanjing area.Methods A total of 525 effective apparently healthy adults were obtained through screening.They were further divided into 4 groups:male and female groups aged 18-49,and male and female groups aged 50-75.Their serum samples were collected,and the UniCel DxI 800 immunoassay system(Beckman)and matching reagents were used to detect serum NT-proBNP levels.The outliers in the detection results were identified,and the 95%reference interval and 90%confidence interval were calculated.Additionally,294 samples were randomly selected from the enrolled population and the levels of se-rum NT-proBNP were detected on the Roche cobas e801 automated electrochemiluminescence analyzer(Roche).The differences and correlations of the detection results between the two systems were compared.Results The test results on the Beckman system showed a right skewed distribution.When analyzing the samples of females aged 18-49 years,a potential outlier with a detection result of 298.4 ng/L was identified.After careful examination,no significant errors were found in the detection process of the sample.The result was confirmed to be valid.Then,the 95%reference interval with a 90%confidence interval was calculated.The final reference intervals ob-tained were≤58.72(49.40,102.00)ng/L for males aged 18-49 years,≤113.72(88.40,141.40)ng/L for females aged 18-49 years,≤97.66(84.00,215.50)ng/L for males aged 50-75 years,≤146.35(126.60,320.80)ng/L for females aged 50-75 years,and≤116.69(103.50,126.70)ng/L for individuals under 75 years,respectively.There were significant differences in the detection data between the Beckman system and Roche system(P＜0.05),and the data between the two systems were positively correlated(r=0.964,P＜0.05),indicating a high reliability of the detection results.Conclusion The reference interval of NT-proBNP accordant with the apparently healthy adults in Nanjing area is preliminarily established,which is suitable for application and promotion in clini-cal diagnosis and treatment.

Objective To establish the reference interval of urine Alzheimer-associated neuronal thread protein(AD7c-NTP)for healthy adults in Mianyang area using the indirect method.Methods The detection results of urine AD7c-NTP from 5 093 healthy in-dividuals were collected from the information management database of Medical Laboratory Department of Sichuan Science City Hospital from March 2017 to March 2022.Skewness-kurtosis and Kolmogorov-Smirnov tests were used to determine whether the data followed a normal distribution.After removing outliers using the Box Plots method,the enrolled subjects were grouped by gender and age.The Mann-Whitney U or Kruska-Wallis H tests were used to analyze the between-group differences of urine AD7c-NTP in healthy individu-als with different genders and ages.The adjacent age groups without statistically significant difference(P＞0.05)were combined,and the indirect method(non-parametric test method)was used to calculate the reference intervals for different gender and age groups.Results Skewness-kurtosis and Kolmogorov-Smirnov tests showed that the data followed a non-normal distribution.After removing 293 outliers using the Box Plots method,a total of 4 800 subjects,including 3 199 males and 1 601 females,were enrolled.The enrolled subjects were grouped by gender and age,and the non-parametric test method were used to establish the reference intervals of urine AD7c-NTP in healthy populations with different genders.The Mann-Whitney U test confirmed that urine AD7c-NTP levels existed gen-der differences(Z=14.09,P＜0.01),and the reference intervals for males and females were≤1.10 ng/mL and≤1.40 ng/mL,re-spectively.There were also statistical differences in urine AD7c-NTP levels among different age groups of the same gender.After combi-ning adjacent age groups without statistically significant difference(P＞0.05),the reference intervals of urine AD7c-NTP in healthy populations with different genders and ages were established by the non-parametric test method,which were≤1.00 ng/mL for male 20-39 years old group,≤1.10 ng/mL for male 40-79 years old group,≤1.60 ng/mL for male≥80 years old group,≤1.30 ng/mL for female 20-69 years old group,and≤1.60 ng/mL for female≥70 years old group,respectively.The established reference intervals of urine AD7c-NTP were further verified by healthy individuals,and the results met the standards.Conclusion The reference intervals of urine AD7c-NTP in healthy populations with different genders and ages in Mianyang area are established successfully using the indi-rect method,which may help to predict the risk of Alzheimer's disease in clinical practice and provide support for the diagnosis and treatment of the disease.

Objective To investigate the epidemiological status and molecular characteristics of diarrheagenic Escherichia coli(DEC)infection in Xiangshan County,providing a basis for its infection surveillance and outbreak control.Methods The real-time fluores-cence quantitative PCR(qRT-PCR)was used to detect virulence genes in isolated E.coli strains.The identified DEC strains were per-formed whole genome sequencing(WGS).The sequencing data were utilized for bacterial resistance gene analysis,multilocus sequence typing(MLST),and core genome multilocus sequence typing(cgMLST).Results A total of 29 DEC strains were isolated,with a detection rate of 9.8%.Enteroaggregative E.coli(EAEC)and enterotoxigenic E.coli(ETEC)were the main strains,accounting for 51.7%and 31.1%,respectively,while enteropathogenic E.coli(EPEC)had a relatively low proportion(17.2%).Drug resistance gene analysis showed that these strains were generally resistant to multiple antibiotics such as fluoroquinolones and carbapenems.MLST typing identified 19 sequence types(STs),including one novel ST type(STcpEC1).Among them,ST117,ST2178,ST31,and ST48 were dominant types.ETEC strains were mainly concentrated in the clonal complex CC-10,while the molecular typing of EAEC and EPEC strains was more dispersed.cgMLST analysis further confirmed the genetic characteristics of different DEC strains.Conclusion DEC infections in Xiangshan County are mainly caused by EAEC,ETEC,and EPEC.These strains have abundant resistance genes.Among them,the molecular typing of EAEC and EPEC is relatively dispersed,while ETEC is concentrated in the clonal complex CC-10.

Objective To investigate the clinical application value of serum exosomal miR-552 and miR-221 in the diagnosis and prog-nosis of gastric cancer.Methods 133 patients diagnosed with gastric cancer in the Department of Gastrointestinal Surgery,Suqian First People's Hospital from January to December 2019 were selected as the gastric cancer group,108 patients with chronic gastritis as the benign lesion group,and 94 healthy individuals undergoing physical examination as the healthy control group.Their blood samples were collected,and serum exosomes were extracted.The expression levels of miR-552 and miR-221 in serum exosomes were detected by real-time fluorescence quantitative PCR(qRT-PCR).The receiver operating characteristics(ROC)curve was used to assess the di-agnostic efficacy of miR-552 and miR-221 alone and in combination for gastric cancer.The correlations of serum exosomal miR-552 and miR-221 levels with the clinicopathological parameters of gastric cancer patients were further analyzed,and the survival curve was used to evaluate the prognosis of gastric cancer patients.Results The expression levels of serum exosomal miR-552 and miR-221 in gastric cancer patients,benign lesion patients,and healthy controls showed statistically significant differences(FmiR-552=296.271,FmiR-221=249.638,P＜0.01)and a decreasing trend.The analysis results of the ROC curve showed that the combined detection of serum exosom-al miR-552 and miR-221 for the diagnosis of gastric cancer had an area under the ROC curve(AUCROC)of 0.901,a sensitivity of 89.50%,and a specificity of 96.90%,all significantly higher than the detection of miR-552(sensitivity of 88.7%and specificity of 87.2%)and miR-221(sensitivity of 83.5%and specificity of 85.1%)alone(P＜0.05).The expression levels of serum exosomal miR-552 and miR-221 were related to tumor size,TNM staging,invasion depth,and lymph node metastasis(P＜0.05).The analysis results of the survival curve revealed that gastric cancer patients with high expression of serum exosomal miR-552 and miR-221 had sig-nificantly shorter survival time than those with low expression(χ2miR-552=30.657,χ2miR-221=38.251,P＜0.05).Conclusion Serum exo-somal miR-552 and miR-221 have high diagnostic efficacy for gastric cancer and may serve as novel biological markers for the diagnosis and prognosis evaluation of gastric cancer.

Objective To investigate the clinical significance and application value of circulating tumor cells(CTCs)detected by fluo-rescence in situ hybridization(FISH)in the diagnosis of breast cancer.Methods A total of 114 breast cancer patients and 30 patients with benign breast lesions admitted to the Department of Thyroid and Breast Surgery,Yixing Hospital Affiliated to Jiangsu University from January 2014 to June 2022 were enrolled.Their peripheral blood cells were enriched by the immunomagnetic bead negative enrich-ment method,and the levels of CTCs were detected by FISH technology.The diagnostic efficacy of CTCs in breast cancer was evaluated by the receiver operating characteristics(ROC)curve,and the correlations of the levels of CTCs with clinicopathological parameters of breast cancer patients were analyzed by the chi-square analysis.Results The detection results of FISH showed that 75 of 114 breast cancer patients were positive for CTCs and 39 were negative.The evaluation results of the ROC curve for the diagnostic efficacy of CTCs in breast cancer showed that when the cut-off value was 2.5 CTCs/mL,its sensitivity and specificity were 52.0%and 89.7%,respec-tively,and the area under the ROC curve(AUCROC)was the best(0.68).There were significant differences in the positive rates of CTCs among different TMN stages and molecular types of breast cancer patients(P＜0.05),but not among different vascular invasion types and immunohistochemical indicators of breast cancer patients(P＞0.05).Conclusion The positive of CTCs detected by FISH may indicate different TNM staging and poor molecular typing.The CTCs is expected to become an important biomarker affecting the clinical decision-making and prognosis evaluation in the treatment of early breast cancer.