Objective:To construct a prognostic model of hepatocellular carcinoma (HCC) with differential expression of autophagy genes.Method:Autophagy genes expression data of HCC and normal liver tissues were obtained from The Cancer Genome Atlas (TCGA) database and The Genotype-Tissue Expression (GTEx) database respectively. The gene expression data from different platforms is normalized into log 2(FPKM value + 1). Differentially expressed autophagy-related genes of HCC were identified by using R program limma package from the TCGA-GTEx combined data set, the criteria of |logFC| > 1 and FDR < 0.05 was deemed to be of statistically significance. The Gene Ontology (GO) analyses and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses were performed by using R program clusterProfiler package, as criteria of P<0.05. Univariate and multivariate Cox proportional hazards regression analyses were performed by using R program survival package to identify the HCC potential prognostic differentially expressed autophagy-related genes. Furthermore, the statistically significant ( P<0.05) autophagy genes in the univariate Cox regression analysis were included in the multivariate Cox regression analysis, and the expression of each differentially expressed autophagy gene and the corresponding regression coefficient coef value based on this, the autophagy gene prognosis model of HCC was constructed: expmRNA1×βmRNA1+ expmRNA2×βmRNA2+ …+ expmRNAn×βmRNAn (exp: gene expression level; β: regression coefficient coef of multivariate Cox regression analysis). Draw the receiver operating characteristic (ROC) curve of the predictive model and calculate the area under curve (AUC) to evaluate the predictive value of the model. Results:The genes expression data and clinical information of 374 HCC samples and 160 normal liver tissue samples were obtained from TCGA and GTEx databases. Total 205 autophagy genes expression data was obtained from the TCGA-GTEx combined sequence. Among them, SPNS1, DIRAS3, TMEM74, NRG2, NRG1, IRGM, IKBKE, NKX2-3, BIRC5, CDKN2A, TP73 are differentially expressed autophagy genes that meet the screening criteria. GO analysis mainly enriched in "regulation of protein serine/threonine kinase activity" , "ErbB 2 signaling pathway" , "protein kinase regulator activity" and "kinase regulator activity" ; KEGG analysis enriched frequently in "EGFR tyrosine kinase inhibitor resistance" , "Hippo signaling pathway" . After integrating and deleting samples with missing survival information, a total of 418 sample expressions were included in the Cox regression analysis. After univariate and multivariate Cox risk regression analysis, the two autophagy genes NRG1 ( HR=1.5565, 95% CI: 1.1793-2.0543) and IKBKE ( HR=1.7502, 95% CI: 1.2093-2.5330) were screened out and a prognostic prediction model was established: (0.44247 × NRG1 expression level) + (0.55977 × IKBKE expression level). The ROC of the prognosis model shows that the AUC of the overall seven-year survival is 0.711. Conclusion:The prognosis model of HCC based on NRG1 and IKBKE has high predictive value for the long-term survival rate of hepatocellular carcinoma patients.

Objective To evaluate the diagnostic value of high frequency ultrasound in sebaceous gland carcinoma of eyelid (SC).Methods The ultrasonic characteristic for 11 cases with eyelid SC were respectively analyzed by using 13 MHz high frequency ultrasound and 22 MHz ultra-high frequency ultrasound.Results Through 13 MHz high frequency ultrasound,in 7 patients who exhibited Pagetoid invasion the lid shin thickness of tumor side displays no significant alteration in a comparison with normal side.Furthermore,the color Doppler flow imaging (CDFI) evealed a branch-like blood flow surrounding the masses in all cases,but the blood flow of seven patients with Pagetoid invasion had no difference compared with the healthy side.On 22 MHz ultra-high frequency ultrasound examination,slit-like low echo was found in 9 ;transition zone of tumor infiltration can be identified in 9 ; the echo of tumors with Pagetoid invasion was lower than the healthy side and the skin thickness of tumors with Pagetoid invasion was thicker (0.6 ±0.1) mm than the healthy side.CDFI revealed that mesh-basket like blood flow was rich in all patients,the small branch blood vessels arrived at subcutaneous,and vasa vasorum were found in some patients.The region with Pagetoid invasion was rich in blood flow.The sonography findings on 13 MHz and 22 MHz high frequency ultrasound examination were compared with chisquare test.There were significant differences on homogeneous echo,slit-like low echo,transition zone of tumor infiltration,infiltration skin thickness,blood distribution,central blood vessels,vasa vasorum,blood flow in the region with Pagetoid invasion (x2 =12.571,15.231,15.231,4.701,22.000,15.231,4.899,10.267,P＜0.05).Conclusions Slit-like low echo in the mass is a main finding of eyelid sebaceous gland cercinoma on the 22 MHz ultra-high frequency ultrasound.The ultra-high frequency ultrasound can accurately reveal the skin depth infiltrated by the eyelid sebaceous gland cercinoma and this method can provide solid guidance for clinical treatment strategies.

Objective:To study the relative proportion of tumorinfiltrating immune cells (TIICs) in colorectal adenocarcinoma (CRC), and to explore the correlation between TIICs and CRC in prognosis and clinical staging.Methods:CRC gene transcriptional expression data and clinical information were obtained from TCGA database. The CIBERSORT software was used to calculate the relative proportions of 22 TIICs in each sample. R software was used to compare the proportion of TIICs between CRC and normal tissues. Single factor survival analysis was performed for each TIICs. Finally, the correlation between each TIICs and CRC clinical stage was studied.Results:A total of 514 gene transcriptional expression data and clinical information were obtained from TCGA database, including 473 CRC and 41normal adjacent tissues.The relative proportions of 22 TIICs in each sample were calculated using the CIBERSORT software "deconvolution method" . In the study, 12 TIICs including naive B cells were found to have statistically significant differences between CRC and normal tissues (all P<0.05). After matching the clinical information of the samples, a total of 222 cases were included in the survival analysis.The relative proportion of each TIICs was arranged in descending order, and all samples were divided into high and low infiltration groups according to the median value. Then, univariate survival analysis was performed for each TIICs, and it was found that memory B cells had a statistically significant effect on the prognosis of CRC ( P<0.05). It was found that the proportion of four types of TIICs, including activated CD 4 memory T cells, in different CRC clinical staging was statistically differe (all P<0.05). Conclusion:TIICs is related to the prognosis and clinical stage of CRC.

Objective To study the correlations between tumor mutation burden (TMB) and the prognosis of hepatocellular carcinoma (HCC) patients,and to investigate the effect of TMB on differential expression genes of HCC and the proportion of invasive immune cells in tumor tissues.Methods The somatic variation data,gene transcriptional expression data and clinical information of HCC patients were obtained from the cancer genome atlas (TCGA) database.The R program language (version 3.6.1)maftools function package was used to analyze the gene mutation data characteristics of the samples.The TMB value of each sample was calculated using the full-exon sequencing data of patients with hepatocellular carcinoma on the VarScan2 platform,sorted by TMB value,and the median value was used to divide all samples into high TMB and low TMB groups.Kaplan-Meier method was used to draw the survival curves of two groups of patients and log-rank test was performed to determine the correlation between tumor mutation load and prognosis.The Limma function package of R language was used to screen the differentially expressed genes between the two groups (FDR =0.05 and logFC =1),and the clusterProfiler function package of R language was used to perform gene ontology (GO) enrichment analysis of the differential genes and kyoto encyclopedia of genes and genomes (KEGG) enrichment analysis (screening criteria were all P ＜ 0.05).Then the CIBERSORT tool was used to compare and analyze the difference in the proportion of invasive immune cells between the two groups.Results A total of 364 patients with HCC from TCGA database were included in the study.Mutations were found in 327 (84％) samples,and there was a synergistic correlation between OBSCN and FLG mutations (P ＜ 0.05),while mutations in CTNNB1 and AXIN1 are mutually exdusive (P ＜ 0.05).A total of 363 patients were included in the TMB survival analysis,sorted by the size of TMB value.All samples were divided into high TMB group (182 cases) and low TMB group (181 cases) with the median value.We found that TMB had no significant effect on the prognosis of HCC patients (P ＞ 0.05).A total of 198 with differentially expressed genes (28 up-regulated genes and 170 down-regulated genes) were screened between the high TMB group and the low TMB group.In GO analysis,it was found that the differentially expressed genes were mainly enriched in extracellular matrix tissues,extracellular structural tissues,extracellular matrix,extracellular matrix containing collagen,extracellular matrix structural components and other functions.In KEGG analysis,differential genes were highly enriched in extracellular matrix receptor interaction pathway and adhesive plaque pathway.In the correlation analysis of the proportion of infiltrating immune cells,CD4 + memory T cells were more infiltrating in the low TMB group (P ＜ 0.05).Monocytes showed a higher degree of infiltration in the high TMB group (P ＜ 0.05).Conclusion There was no correlation between TMB and the prognosis of HCC patients.TMB has significant influence on the differential expression genes of HCC and the proportion of invasive immune cells in tumor tissues.

Objective To investigate the effect of docosahexaenoic acid (DHA) on the sensitivity of esophageal carcinoma cell line EC9706 to cisplatin and the mechanism behind this effect.Methods EC9706 cells were randomly divided into 5 groups:control group,DHA group,cisplatin (DDP) group,DHA+DDP group and endoplasmic reticulum stress (ERS) activation tunicamycin (TM) group (DHA+DDP+TM group).MTT method was used to evaluate inhibition ratio of cell proliferation.The apoptotic ratio was examined by flow eytometry.Western blot was used to detect the protein expressions of apoptosis cytokines (caspase-3 and Bcl-2) and ERS cytokines [glucose-regulated protein 78 (GRP78) and inositol-requiring enzyme 1 (IRE-1)].Results DHA causes concentration-dependent and time-dependent inhibition of the proliferation of EC9706 cells (P=0.00).DHA significantly enhanced the sensitivity of esophageal carcinoma cell line EC9706 to cisplatin.Compared to DDP treatment alone,the inhibition ratio [(60.19±5.05)％ vs.(36.72±3.52)％,P=0.02] and apoptotic ratio [(54.88±4.94)％ vs.(39.74±4.64)％,P=0.03] of EC9706 cells were enhanced by DHA+DDP treatment.Western blot showed that the expression of apoptotic factor caspase-3 protein was increased by DHA+DDP treatment.Meanwhile,the protein expressions of anti-apoptotic factor (Bcl-2) and ERS-related factors (GRP78 and IRE-1) were significantly inhibited by DHA+DDP treatment (P=0.01).However,the salutary effects of DHA were reversed by ERS activation tunicamycin.Conclusion DHA enhances the sensitivity of esophageal carcinoma cell line EC9706 to cisplatin,the mechanism of which may be the suppression of ERS response.

The incidence of non-alcoholic fatty liver disease (NAFLD) is increasing year by year, and it has become a serious public health problem that threatens human health.NAFLD patients can progress from simple steatosis to non-alcoholic steatohepatitis (NASH), liver fibrosis, and even cirrhosis without intervention. Therefore, early diagnosis and treatment are particularly important.Liver biopsy is the gold standard for diagnosing NAFLD, but its limitations such as invasion, bleeding, and high cost limit its widespread clinical application. There is an urgent need for sensitive and convenient non-invasive diagnostic methods to achieve early diagnosis and treatment of NAFLD.This article provides a review of the progress in serological diagnostic methods for NAFLD both domestically and internationally in recent years.

Objective:The incidence of early-onset colorectal cancer (EOCRC) is increasing globally; however, the molecular characteristics and prognosis of sporadic EOCRC are unclear. In this systematic review and meta-analysis, we aimed to investigate the incidence of gene mutations and their association with cancer survival in sporadic EOCRC, focusing on six common gene mutations ( TP53, BRAF, KRAS, NRAS, PTEN, and APC). Methods:Ovid Embase and Ovid Medline electronic databases were searched for studies involving patients with sporadic EOCRC (i.e., diagnosed with colorectal cancer before the age of 50 years and with no evidence of hereditary syndromes predisposing to colorectal cancer). The included articles were evaluated using quality assessment tools. Meta-analysis was performed using random-effects and fixed-effects models. Cochran's Q statistic and the I2 index were used to assess heterogeneity. The incidence of the six common gene mutations listed above in sporadic EOCRC and their association with cancer survival were evaluated.Results:(1) Incidence of specific gene mutations in sporadic EOCRC. A total of 34 articles were included in this meta-analysis. The incidence of APC gene mutation was 36% (from 13 articles, 95%CI: 19%-55%, P=0.043); of KRAS gene mutation 30% (from 26 articles, 95%CI: 24%-35%, P=0.190); of BRAF gene mutation 7% (from 18 articles, 95%CI: 5%-11%, P=0.422); of NRAS gene mutation 4% (from five articles, 95%CI: 3%-5%, P=0.586); of PTEN gene mutation 6% (from six articles, 95%CI: 4%-10%, P=0.968); and of TP53 gene mutation 59% (from 13 articles, 95%CI: 49%-68%, P=0.164). (2) Association between gene mutations and survival in sporadic EOCRC . A total of six articles were included in this meta-analysis. Compared with wild-type BRAF, mutant BRAF was significantly associated with increased overall mortality risk in patients with EOCRC (pooled HR=2.85, 95%CI: 1.45-5.60, P=0.002). Subgroup analysis showed that the incidence of BRAF gene mutation was higher in Eastern than in Western countries, whereas the incidence of TP53, KRAS, NRAS, and APC gene mutations was lower. There was no significant difference in the incidence of PTEN gene mutation between different regions. Conclusion:Compared with colorectal cancer occurring in the general population, the incidence of APC and KRAS mutations is lower in EOCRC, whereas the incidence of TP53 mutation remains consistent. BRAF mutation is associated with increased overall mortality risk in patients with EOCRC.

Objective:The incidence of early-onset colorectal cancer (EOCRC) is increasing globally; however, the molecular characteristics and prognosis of sporadic EOCRC are unclear. In this systematic review and meta-analysis, we aimed to investigate the incidence of gene mutations and their association with cancer survival in sporadic EOCRC, focusing on six common gene mutations ( TP53, BRAF, KRAS, NRAS, PTEN, and APC). Methods:Ovid Embase and Ovid Medline electronic databases were searched for studies involving patients with sporadic EOCRC (i.e., diagnosed with colorectal cancer before the age of 50 years and with no evidence of hereditary syndromes predisposing to colorectal cancer). The included articles were evaluated using quality assessment tools. Meta-analysis was performed using random-effects and fixed-effects models. Cochran's Q statistic and the I2 index were used to assess heterogeneity. The incidence of the six common gene mutations listed above in sporadic EOCRC and their association with cancer survival were evaluated.Results:(1) Incidence of specific gene mutations in sporadic EOCRC. A total of 34 articles were included in this meta-analysis. The incidence of APC gene mutation was 36% (from 13 articles, 95%CI: 19%-55%, P=0.043); of KRAS gene mutation 30% (from 26 articles, 95%CI: 24%-35%, P=0.190); of BRAF gene mutation 7% (from 18 articles, 95%CI: 5%-11%, P=0.422); of NRAS gene mutation 4% (from five articles, 95%CI: 3%-5%, P=0.586); of PTEN gene mutation 6% (from six articles, 95%CI: 4%-10%, P=0.968); and of TP53 gene mutation 59% (from 13 articles, 95%CI: 49%-68%, P=0.164). (2) Association between gene mutations and survival in sporadic EOCRC . A total of six articles were included in this meta-analysis. Compared with wild-type BRAF, mutant BRAF was significantly associated with increased overall mortality risk in patients with EOCRC (pooled HR=2.85, 95%CI: 1.45-5.60, P=0.002). Subgroup analysis showed that the incidence of BRAF gene mutation was higher in Eastern than in Western countries, whereas the incidence of TP53, KRAS, NRAS, and APC gene mutations was lower. There was no significant difference in the incidence of PTEN gene mutation between different regions. Conclusion:Compared with colorectal cancer occurring in the general population, the incidence of APC and KRAS mutations is lower in EOCRC, whereas the incidence of TP53 mutation remains consistent. BRAF mutation is associated with increased overall mortality risk in patients with EOCRC.

Objective:To investigate the efficacy and diagnostic accuracy of changes in cytokine levels before and after non-biological artificial liver (referred to as ABL) treatment in patients with acute-on-chronic liver failure (ACLF) in order to establish a basis for treatment timing selection and short-term (28d) prognosis.Methods:90 cases diagnosed with ACLF were selected and divided into a group receiving artificial liver treatment (45 cases) and a group not receiving artificial liver treatment (45 cases). Age, gender, first routine blood test after admission, liver and kidney function, and procalcitonin (PCT) of the two groups were collected. The 28-day survival of the two groups was followed-up for survival analysis. The 45 cases who received artificial liver therapy were further divided into an improvement group and a deterioration group according to the clinical manifestations before discharge and the last laboratory examination results as the efficacy evaluation indicators. Routine blood test, coagulation function, liver and kidney function, PCT, alpha fetoprotein (AFP), β-defensin-1 (HBD-1), 12 cytokines and other indicators were analyzed and compared. A receiver operating characteristic curve (ROC curve) was used to analyze the diagnostic efficacy of the short-term (28 d) prognosis and an independent risk factors affecting the prognosis of ACLF patients. According to different data, Kaplan-Meier method, log-rant test, t-test, Mann-Whitney U test, Wilcoxon rank-sum test, χ2 test, Spearman rank correlation analysis and logistic regression analysis were used for statistical analysis. Results:The 28-day survival rate was significantly higher in ACLF patients who received artificial liver therapy than that of those who did not receive artificial liver therapy (82.2% vs. 61.0%, P<0.05). The levels of serum HBD-1, alpha interferon (IFN-α) and interleukin-5 (IL-5) after artificial liver treatment were significantly lower in ACLF patients than those before treatment ( P<0.05), while liver and coagulation function were significantly improved compared with those before treatment ( P<0.05), and there was no statistically significant difference in other serological indexes before and after treatment ( P＞0.05). Before artificial liver treatment, serum HBD-1 and INF-α levels were significantly lower in the ACLF improvement group than in the deterioration group ( P<0.05) and were positively correlated with the patients' prognosis (deteriorating) ( r=0.591, 0.427, P<0.001, 0.008). The level of AFP was significantly higher in the improved ACLF group than that in the deterioration group ( P<0.05), and was negatively correlated with the prognosis (deteriorating) of the patients ( r=-0.557, P<0.001). Univariate logistic regression analysis showed that HBD-1, IFN-α and AFP were independent risk factors for the prognosis of ACLF patients ( P=0.001, 0.043, and 0.036, respectively), and that higher HBD-1 and IFN-α levels were associated with lower AFP level and a deteriorating prognosis. The area under the curve (AUC) of HBD-1, IFN-α, and AFP for short-term (28d) prognostic and diagnostic efficacy of ACLF patients was 0.883, 0.763, and 0.843, respectively, and the sensitivity and specificty was 0.75, 0.75, and 0.72, and 0.84, 0.80, and 0.83, respectively. The combination of HBD-1 and AFP had further improved the diagnostic efficiency of short-term prognosis of ACLF patients (AUC=0.960, sensitivity and specificity: 0.909 and 0.880 respectively). The combination of HBD-1+IFN-α+AFP had the highest diagnostic performance, with an AUC of 0.989, sensitivity of 0.900, and specificity of 0.947. Conclusion:Artificial liver therapy can effectively improve the clinical symptoms and liver and coagulation function of patients with ACLF; remove cytokines such as HBD-1, IFN-α, and IL-5 in patients with liver failure; delay or reverse the progression of the disease; and improve the survival rate of patients. HBD-1, IFN-α, and AFP are independent risk factors affecting the prognosis of ACLF patients, which can be used as biological indicators for evaluating the short-term prognosis of ACLF patients. The higher the level of HBD-1 and/or IFN-α, the higher the risk of disease deterioration. Therefore, artificial liver therapy should be started as soon as possible after the exclusion of infection. In diagnosing the prognosis of ACLF, HBD-1 has higher sensitivity and specificity than IFN-α and AFP, and its diagnostic efficiency is greatest when combined with IFN-α and AFP.

Objective:To explore the diagnostic value of serum C-C chemokine ligand 5 (CCL5) in assessing the degree of liver fibrosis in patients with metabolic-associated fatty liver disease (MAFLD).Methods:71 MAFLD patients who visited the Department of Integrated Traditional and Western Medical Hepatology, Third Hospital of Hebei Medical University, and underwent liver biopsy histopathology examinations between October 2021 and June 2023 were selected for diagnostic testing. Simultaneously, 71 healthy subjects who underwent physical examinations at the physical examination center of the hospital were selected as the control group. Serum CCL5 levels were detected using an enzyme-linked immunosorbent assay (ELISA). Routine blood tests, liver and kidney function tests, and other tests were conducted to analyze the expression level of CCL5 and its correlation with the above indicators. The aspartate aminotransferase/platelet ratio index (APRI) and fibrosis 4 index (FIB-4) were calculated. SPSS 26.0 software was used for statistical analysis. The area under the receiver operating characteristic curve (ROC) was used to evaluate the diagnostic efficacy of CCL5 for the degree of liver fibrosis in MAFLD. The combined diagnostic efficacy of APRI and FIB-4 was further analyzed for the degree of liver fibrosis in MAFLD.Results:The expression level of serum CCL5 gradually increased with the increase in liver fibrosis stage in patients with MAFLD, and the difference was statistically significant ( P<0.05). The AUC value of serum CCL5 for diagnosing significant liver fibrosis in MAFLD patients was 0.775, with a sensitivity of 65.7%, a specificity of 80.6%, and an optimal cutoff value of 49.845 ng/ml. The CCL5 and FIB-4 combination had the highest diagnostic value for significant liver fibrosis in patients with MAFLD, with an AUC of 0.802, a sensitivity of 91.4%, and a specificity of 61.1%. Conclusion:CCL5 has a high diagnostic value for significant liver fibrosis in MAFLD patients. Therefore, it is expected to become a non-invasive diagnostic marker for assessing the degree of liver fibrosis in MAFLD patients.