Background/Aims: Liver transplantation (LT) following total hepatectomy is a life-saving treatment for hepatocellular carcinoma (HCC). The HCC recurrence after LT hinders the effectiveness of the procedure. The objective of this study is to develop a pre-operative risk stratification model based on a liquid biopsy. Methods: We conducted a comprehensive multi-omics study of 260 HCC patients from three centers, including clinical data, low-coverage whole-genome sequencing of cell-free DNA (cfDNA) from plasma, as well as whole-exome, single-nucleus RNA, and spatial transcriptomics from matched tumor and non-tumor tissues. Results: We identified cfDNA-derived copy number alteration (CNA) signatures associated with post-transplant recurrence. By integrating cfDNA-derived CNA profiles with single-cell transcriptomic data, we traced recurrence-associated cfDNA to a distinct subpopulation of malignant cells within the primary tumor. These cells were embedded in a pro-metastatic microenvironment of specialized endothelial subtypes and cancer-associated fibroblasts. Notably, most recurrence-associated lesions were detectable in cfDNA prior to liver transplantation (LT). Building on these insights, we developed the ZJU Criteria based on CNA fragments and tumor markers, a pre-LT risk prediction tool that integrates conventional clinical factors with cfDNA-derived CNA signatures, and validated it using internal and independent external cohorts. Conclusion Our findings suggest that post-transplant recurrence commonly originates from advanced subclones that emerge late during tumor evolution. The ZJU Criteria provides an accurate, non-invasive strategy that significantly improves pre-LT risk stratification and clinical decision-making for patients with HCC.

Objective:To investigate the influence of cytochrome P450 2A6 (CYP2A6) gene polymorphisms on liver function injury in patients with hyperthyroidism treated with methimazole.Methods:The study selected 90 patients with hyperthyroidism who were treated with methimazole in the Department of Thyroid Surgery at the First Affiliated Hospital of Zhengzhou University from Sept. 2023 to Aug. 2024 as the research subjects. Based on the occurrence of liver injury, they were divided into a liver injury group ( n=36) and a non-liver injury group (n=54). Peripheral blood DNA was extracted from the patients, and the CYP2A6 gene genotypes (rs8192725, rs8192720, and rs28399433) were detected using the polymerase chain reaction (PCR) amplification method. The association between CYP2A6 gene polymorphisms and liver injury induced by methimazole treatment in hyperthyroidism was analyzed. Results:The comparison of genotype distribution frequencies at the rs8192725 locus between the liver injury group and the non-liver injury group showed a statistically significant difference ( P<0.05). The AG and GG genotypes at the rs8192725 locus were protective factors against liver injury in patients with hyperthyroidism (AG vs. AA, OR: 0.21; 95% CI: 0.08-0.57; P<0.05; GG vs. AA, OR: 0.24; 95% CI: 0.06-0.89; P<0.05; AG+GG vs. AA, OR: 0.22; 95% CI: 0.09-0.54; P<0.05). The frequency of the G allele of rs8192725 in the liver injury group was significantly lower than that in the non-liver injury group (G vs. A, OR: 0.36; 95% CI: 0.19-0.70; P<0.05), indicating that it is a protective factor for liver injury in hyperthyroid patients receiving methimazole treatment. Conclusions:The CYP2A6 gene polymorphism at the rs8192725 locus is associated with the occurrence of liver injury in patients with hyperthyroidism treated with methimazole. The G allele may be a protective factor against liver injury in patients with hyperthyroidism, suggesting that individualized treatment plans can be developed based on the patient's genotype.

Objective To establish an animal model of hand,foot,and mouth disease(HFMD)in Syrian hamsters coinfected with coxsackievirus A6(CVA6)and coxsackievirus B1(CVB1).Methods 42 Syrian hamsters were divided into a CVA6 infection group,CVB1 infection group,CVA6 and CVB1 coinfection group and control group.A HFMD model was established by nasal instillation of virus solution and phosphate-buffered saline.Clinical and physiological indicators and detoxification status were monitored and recorded for 15 d,and animals were selected on day 7(D7)after infection for histopathology and viral antigen and nucleic acid testing.Results Hamsters in the single-infection and coinfection groups showed clinical symptoms similar to human HFMD.White blood cell,neutrophil,and lymphocyte result were characteristic of viral infection.Both viral nucleic acids were detected in throat swabs,feces,blood,and tissues and both viruses were isolated from fecal samples.Pathological damage and positive co-localization of CVA6 and CVB1 viral antigen proteins and nucleic acids were found in brain and other tissues.Conclusions Nasal instillation of a CVA6 and CVB1 mixture can successfully coinfect Syrian hamsters,replicate herpes infection similar to human HFMD,and cause pathological viral myocarditis and encephalitis damage.The result showed that the coinfection group was more seriously affected than the single-infection group,with worse clinical symptoms,increased viral replication,and obvious tissue pathological damage.This study provides a reference for further basic and clinical research into human enterovirus coinfection.

Objective:To investigate the influence of cytochrome P450 2A6 (CYP2A6) gene polymorphisms on liver function injury in patients with hyperthyroidism treated with methimazole.Methods:The study selected 90 patients with hyperthyroidism who were treated with methimazole in the Department of Thyroid Surgery at the First Affiliated Hospital of Zhengzhou University from Sept. 2023 to Aug. 2024 as the research subjects. Based on the occurrence of liver injury, they were divided into a liver injury group ( n=36) and a non-liver injury group (n=54). Peripheral blood DNA was extracted from the patients, and the CYP2A6 gene genotypes (rs8192725, rs8192720, and rs28399433) were detected using the polymerase chain reaction (PCR) amplification method. The association between CYP2A6 gene polymorphisms and liver injury induced by methimazole treatment in hyperthyroidism was analyzed. Results:The comparison of genotype distribution frequencies at the rs8192725 locus between the liver injury group and the non-liver injury group showed a statistically significant difference ( P<0.05). The AG and GG genotypes at the rs8192725 locus were protective factors against liver injury in patients with hyperthyroidism (AG vs. AA, OR: 0.21; 95% CI: 0.08-0.57; P<0.05; GG vs. AA, OR: 0.24; 95% CI: 0.06-0.89; P<0.05; AG+GG vs. AA, OR: 0.22; 95% CI: 0.09-0.54; P<0.05). The frequency of the G allele of rs8192725 in the liver injury group was significantly lower than that in the non-liver injury group (G vs. A, OR: 0.36; 95% CI: 0.19-0.70; P<0.05), indicating that it is a protective factor for liver injury in hyperthyroid patients receiving methimazole treatment. Conclusions:The CYP2A6 gene polymorphism at the rs8192725 locus is associated with the occurrence of liver injury in patients with hyperthyroidism treated with methimazole. The G allele may be a protective factor against liver injury in patients with hyperthyroidism, suggesting that individualized treatment plans can be developed based on the patient's genotype.

Objective:To establish the quality standard of the standard decoction of wine-processed Coptidis Rhizoma by studying the extraction rate, fingerprint and component quantitative analysis.Methods:ccording to the Technical Requirements for Quality Control and Standard Formulation of Chinese Medicine Formula Granules, 15 batches of the standard decoction of wine-processed Coptidis Rhizoma were prepared, and the paste rate was determined; HPLC fingerprints of 15 batches of standard decoction of wine-processed Coptidis Rhizoma were established, and evaluated by combining similarity evaluation, clustering analysis, principal component analysis and orthogonal partial least squares discriminant analysis; the contents of berberine, epiberberine, pamadine, and safranine in the samples of the 15 batches were determined and analyzed their transfer rates.Results:A total of 15 batches of standard decoction samples were calibrated with 11 common peaks, referring to the recognition of 8 components. The similarity between the samples and the control product was greater than 0.900; the clustering analysis could cluster the 15 batches of samples into 2 classes; the results of the principal component analysis showed that the cumulative variance contribution rate of the 3 principal component factors was 89.388%; the OPLS-DA screened out the 3 components of the quality difference; the 15 batches of samples out of the paste rate was 15.7% -20.8%, and the mass fractions of berberine, epiberberine, safranine, and palmatine were 18.47%-24.38%, 2.82%-3.49%, 5.08%-6.69%, and 4.84%-6.68%, respectively, with transfer rates of 41.7%-61.7%, 46.9%-68.7%, 39.8%-61.5%, and 43.8%-65.2%.Conclusion:The fingerprint and content determination method established in this study is accurate, stable, simple, and can be used for the quality control and evaluation of the standard decoction of wine-processed Coptidis Rhizoma.

Objective To establish an animal model of hand,foot,and mouth disease(HFMD)in Syrian hamsters coinfected with coxsackievirus A6(CVA6)and coxsackievirus B1(CVB1).Methods 42 Syrian hamsters were divided into a CVA6 infection group,CVB1 infection group,CVA6 and CVB1 coinfection group and control group.A HFMD model was established by nasal instillation of virus solution and phosphate-buffered saline.Clinical and physiological indicators and detoxification status were monitored and recorded for 15 d,and animals were selected on day 7(D7)after infection for histopathology and viral antigen and nucleic acid testing.Results Hamsters in the single-infection and coinfection groups showed clinical symptoms similar to human HFMD.White blood cell,neutrophil,and lymphocyte result were characteristic of viral infection.Both viral nucleic acids were detected in throat swabs,feces,blood,and tissues and both viruses were isolated from fecal samples.Pathological damage and positive co-localization of CVA6 and CVB1 viral antigen proteins and nucleic acids were found in brain and other tissues.Conclusions Nasal instillation of a CVA6 and CVB1 mixture can successfully coinfect Syrian hamsters,replicate herpes infection similar to human HFMD,and cause pathological viral myocarditis and encephalitis damage.The result showed that the coinfection group was more seriously affected than the single-infection group,with worse clinical symptoms,increased viral replication,and obvious tissue pathological damage.This study provides a reference for further basic and clinical research into human enterovirus coinfection.

Objective To compare the effect of extreme learning machine(ELM)vs logistic regression analysis on prediction of the risk to PICC-related central line associated bloodstream infections(PICC-CLABSI)in cancer patients.Methods Clinical data of 1,146 patients who received PICC,from January 2019 to March 2023,in the Department of Oncology of a ⅢA hospital were analysed.A total of 786 patients who received PICC between January 2019 and December 2021 were assigned to the modelling group,and the rest of 360 patients who received PICC between January 2022 and March 2023 were assigned to the validation group.The risk prediction model was established based on the data of modelling group analysed by Chi-square test,and then by the binary logistic regression to determine the statistically significant variables.Based on the analyses of the two models,a nomogram was plotted to evaluate the fitting and predictive effectiveness.Performance of the two models were evaluated using Hosmer-Lemeshow test as well as the area under the curve(AUC)of receiver operating characteristic(ROC).Risk factors identified by the logistic regression and the PICC-CLABSI risks were used as input and output parameters respectively,to establish an ELM prediction model.The two models were compared in terms of predictive effectiveness using the data of the validation group.Results History of diabetes mellitus,frequency of chemotherapy(≥3 times),maintenance cycle(>7 days),maintenance site(out of hospital),white blood cell count(<3.5×109/L),and albumin(<40g/L)were risk factors for PICC-CLABSI in cancer patients.The logistic regression model demonstrated a good predictability by Hosmer-Lemeshow test(χ 2=5.201,P=0.736),with an AUC-ROC of 0.860(95%CI:0.799～0.922),sensitivity at 0.893,specificity at 0.704 and accuracy at 72.8%.The ELM prediction model exhibited a determination coefficient of 0.823 and mean squared error of 0.051,with a fitting rate at 74.5%,hence it indicated a good predictive power.The ELM model showed a superior predictive power than the logistic regression model.Conclusion The ELM model and logistic regression model,based on logistic regression analysis,offers higher prediction accuracy.It provides valuable guidance to healthcare providers in identification of high risks of PICC-CLABSI for cancer patients.

Objective:To establish HPLC chromatogram for Aspongopus; To determine 8 nucleoside components of uracil, adenine, uridine, uric acid, hypoxanthine, adenosine, xanthine and canine quinolinic acid; To provide reference for quality control and evaluation.Methods:The Agilent ZORBAX SB-Aq chromatographic column (4.6 mm×250 mm, 5 μm) was used for gradient elution with mobile phases consisting of a methanol (A) and 0.05% phosphoric acid (B). The column temperature was 25 ℃, the flow rate was 0.8 ml/min, and the detection wavelength was 254 nm. HPLC chromatograms for Aspongopus were established and the contents of 8 components were determined.Results:The characteristic chromatogram of 15 batches aspongopus herbs was established. A total of 10 common characteristic peaks were identified and 8 were identified. The similarity between the characteristic chromatogram of samples and the control chromatogram was 0.969-0.997. The content determination showed that the linear range of uracil, adenine, urin, uric acid, hypoxanthine, adenosine, xanthine and xanuric acid was among 0.002 0-0.644 0, 0.001 4-0.448 0, 0.001 0-1.257 0, 0.005 4-6.221 0, 0.001 0-0.724 0, 0.001 0-0.644 0, 0.002 0-1.113 0, 0.003 8-2.059 0 μg, respectively, with a good linear relationship ( r≥0.999); the repeatability and stability of RSD were <2.0%, and the average sampling recovery rate was between 99.36% and 103.40%. Conclusion:The characteristic chromatogram and content determination method established in this study are simple, reliable, reproducible and accurate, and can be used for the qualitative and quantitative analysis of Aspongopus and can provide a reference for the quality evaluation method of the Aspongopus.

Objective:To investigate the changes of ROS/TXNIP/NLRP3 signaling pathway in pyroptosis of human embryonic trophoblast cells induced by high glucose.Methods:Human embryonic trophoblast cells were cultured in vitro to establish high glucose injury model, and they were randomly divided into control group, high glucose (HG) group and HG + ROS inhibitor N-acetyl-L-cysteine (HG + NAC) group. MTT assay was used to detect the cell survival rate. The level of ROS in each group was detected by dihydroethidine ROS fluorescence probe. Expression of TXNIP and NLRP3 mRNA was detected by real-time quantitative PCR (RT-qPCR). Western blot analysis was used to detect the expression levels of TXNIP, NLRP3, Caspase-1, interleukin (IL) -1β, tumor necrosis factor-α (TNF-α) and GSDMD proteins. In addition, pyroptosis was detected by flow cytometry.Results:The optimal glucose concentration for high glucose-induced injury of human embryonic trophoblast cells was 30 mmol/L. Compared with the control group (96.27±3.10) %, the survival rate of human embryonic trophoblast cells in HG group (55.44±2.15) % was significantly lower ( P<0.05), while the fluorescence intensity (ROS level) of 7 'dichlorofluorescein (DCF), the expression levels of TXNIP and NLRP3 proteins, the number of pyroptosis, expression levels of Caspase-1, GSDMD, IL-1β and TNF-α proteins were significantly higher ( P<0.05) ; Compared with HG group, the survival rate of human embryonic trophoblast cells in HG+NAC group (84.75±2.33) % was significantly higher ( P<0.05), the fluorescence intensity (ROS level) of DCF, the expression levels of TXNIP and NLRP3 proteins, the number of pyroptosis, and expression levels of Caspase-1, GSDMD, IL-1β and TNF-α proteins were significantly lower ( P<0.05) . Conclusion:Inhibition of ROS level in human embryonic trophoblast cells induced by high glucose may promote cell proliferation and reduce the occurrence of pyroptosis by inhibiting TXNIP/NLRP3 signaling pathway.

Objective:To analyze the detective value of placental tissue resistin, human lipid carrier protein (LCN) and blood glucose and lipid metabolism in pregnant women with gestational diabetes mellitus (GDM) complicated with preeclampsia (PE) , providing guidance for the early treatment of GDM complicated with preeclampsia.Methods:96 pregnant women with GDM complicated with PE (GDM-PE group) admitted to Yantai Yantaishan Hospital from Jan. 2017 to Jan. 2020 were selected and retrospectively studied. According to the ratio of 2:1, the pure GDM pregnant women (GDM group) and 48 normal pregnant women (the control group) were selected. The placenta tissue resistin and LCN levels were determined by immunohistochemistry. Blood samples were collected to determine the glucose and lipid metabolism. The pregnancy outcomes of each group were compared and the relationship between resistin, LCN, glucose and lipid metabolism and GDM complicated with PE was analyzed.Results:Fasting blood-glucose (FBG) was (4.57±0.66) mmol/L in GDM group and (5.23±0.61) mmol/L in GMD-PE group. FINS (11.97±1.5) mIU/L, (15.12±3.52) mIU/L were higher than those of control group (4.11±0.23) mmol/L, (6.75±1.34) mIU/L ( P<0.05) . FBG, FINS, glycosylated hemoglobin (HbA1c) in GDM-PE group were higher than those in GDM group. TC) (6.71±1.63) mmol/L, triglyceride, TG (6.59±0.87) mmol/L was higher than that of control group (5.87±0.73) mmol/L, (4.57±0.59) mmol/L and GDM group (6.02±1.55) mmol/L, (4.71±0.63) mmol/L ( P<0.05) . high density lipoprotein cholesterol (HDL-C) (1.21±0.34) was lower than that of control group (1.54±0.39) and GDM group (1.55±0.43) ( P<0.05) . The positive rates of resistin 85.42%, 60.42%, LCN 81.25%, 56.25% in GDM-PE group and GDM group were higher than those in control group 39.58%, 31.25% ( χ2=32.096, 4.167; 34.975, 6.095, both P<0.05) . The positive rates of resistin and LCN in GDM-PE group were higher than those in GDM group ( χ2=11.322, 11.257, both P<0.01) . The gestational age of delivery in GDM-PE group was (37.11±2.06) weeks earlier than that in GDM group (38.21±1.75) weeks and control group (38.36±1.42) weeks ( F=9.836, P<0.05) . The birth weight of neonates (2 905.45±356.79) g was lower than that of control group (3 321.52±366.46) g and GDM group (3 425.14±269.87) g ( F=46.606, P<0.05) . Postpartum blood loss (415.34±126.75) ml was significantly higher than that of GDM group (338.65±105.63) ml and control group (298.42±75.26) ml ( F=19.932, P<0.05) . The preterm birth rate of 20.83% was higher than that of the GDM group (8.33%) and the control group (4.17%) ( χ2=9.075, P<0.05) . The postpartum blood loss of the GDM group was higher than that of the control group ( t=-2.148, P<0.05) . The incidences of fetal distress, premature rupture of membranes, fetal growth restriction and postpartum hemorrhage in GDM-PE group were higher than those in control group ( χ2=4.571, 6.867, 5.941, 5.123, P<0.05) . The protein expressions of resistin and LCN in placenta of pregnant women with GDM-PE were positively correlated with FBG, FINS, TC and TG ( r=0.517, 0.463, 0.559, 0.521, 0.485, 0.497, 0.557, 0.571, P<0.05) . Was negatively correlated with HDL-C ( r=-0.317, -0.357, P<0.05) . Conclusions:The positive rate of resistin and LCN in the placenta tissue of pregnant women with GDM complicated with PE is higher than that of GDM and normal pregnant women, their disorder of glucose and lipid metabolism is more obvious, and the incidence of adverse maternal and infant outcomes is higher. It is speculated that resistin and LCN may synergistically affect the metabolism of glucose and lipids causing adverse pregnancy outcomes in GDM complicated with PE.