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 establish a mouse model of spinal cord injury(SCI),and to investigate the effect of electroacupuncture(EA)intervention on cell apoptosis after acute SCI and its mechanism.Methods:Female C57BL/6 mice were used to establish a model of SCI,and after successful modeling,the mice were randomly divided into SCI group,EA group,and Rosiglitazone group(R group);a sham-operation group(Sham group)was also established.After successful modeling,the mice in the EA group were given EA at bilat-eral Jiaji points and Zusanli once a day for 14 days,those in the R ture,and the number of surviving cells.The EA group and the R group had a significant reduction in the expression of caspase-3 and significant increases in the expression of PPARγ and CD36,and the EA group had significant reductions in the expression of Hba-a1 and Hbb-bt.In addition,RNA-Seq and TMT/iTRAQ techniques,significant analysis,Venn analysis,and dual-omics analysis identi-fied Hba-a1 and Hbb-bt as the target genes of EA.The KEGG pathway enrichment analysis showed that EA had a significant effect on the PPAR signaling pathway.Conclusion:By regulating the PPARγ-CD36 signaling pathway,EA can promote the clearance of Hba-a1 and Hbb-bt after SCI,reduce the expression level of caspase-3,alleviate cell apoptosis,and facilitate the recovery of spinal cord nerve function.

Objective To summarize and evaluate the characteristics of current recurrent spontaneous abortion(RSA)animal models at home and abroad,and to provide reference and guidance for the standardized preparation of RSA models.Methods"Recurrent spontaneous abortion"and"animal model"were used as co-keywords in CNKI,Wanfang,VIP,PubMed and Web of Science databases to search the RSA animal experimental literature,covering the period up to January 20,2024,and a total of 1 411 articles were collected.The analysis focused on construction methods and essential elements of RSA animal models,the modeling process and result evaluation,as well as the application of these models in pharmacological and pharmacodynamic research.An Excel table was established for systematic analysis and discussion.Results A total of 138 experimental studies were obtained after screening.In constructing RSA animal models,immunological models were the most widely used in Western medicine(96.92％),with the Clark model being the main one(92.31％).In traditional Chinese medicine(TCM)models,70.00％were kidney deficiency-luteal inhibition-syndrome combination models,20.00％were kidney deficiency and blood stasis models,and 10.00％were deficiency-heat syndrome models.Most animals were selected at 6-8 weeks(33.86％)and 8 weeks(32.28％)of age.The majority of animals were paired for mating at 18:00 on the day of cage pairing.In 81.03％of literatures,vaginal plugs were checked once the following morning,with 8:00 being the most common time(17.02％).The most commonly used drug administration cycle was 14 days of continuous gavage after pregnancy.Among the tested drugs,Western drugs were mainly protein-based(29.17％),while TCM drugs were mainly TCM decoction(81.11％).The most frequently used methods for detecting indicators included visual observation of embryos(22.54％),western blot(15.96％),PCR(13.58％),ELISA(12.91％),HE staining(10.80％)and immunohistochemistry(9.39％).Conclusion The etiology of RSA is complex,and corresponding animal models should be established based on different etiologies.Clark model is commonly used in the construction of Western medicine model,while the kidney deficiency-luteal inhibition-syndrome combination model is predominant in TCM.RSA animal model is widely used in related research,but systematic evaluation needs to be strengthened.

Cross-Sectional Studies ; Urodynamics ; China

Objective: : This study aimed to investigate the changes and significance of microRNA155 levels in serum of patients with cerebral small vessel disease (CSVD). Methods: : Thirty patients with CSVD who met the inclusion criteria were selected and divided into eight patients with lacunar infarction (LI) group and 22 patients with multiple lacunar infarction (MLI) combined with white matter lesions (WML) group according to the results of head magnetic resonance imaging (MRI). Thirty samples from healthy volunteers without abnormalities after head MRI examination were selected as the control group. The levels of serum microRNA155 in each group were determined by real-time polymerase chain reaction, and the correlation between microRNA155 in the serum of patients with CSVD and the increase of imaging lesions was analyzed by Spearman correlation analysis. Results: : Compared with the control group, the serum microRNA155 level in the LI group, MLI combined with WML group increased, the difference was statistically significant (p<0.05); serum microRNA155 level was positively correlated with the increase of imaging lesions (p<0.05). Conclusion : The change of serum microRNA155 level in patients with CSVD may be one of its self-protection mechanisms, and the intensity of this self-protection mechanism is positively correlated with the number of CSVD lesions.

Objective To observe and evaluate the effects of metformin and pioglitazone on blood glucose,insulin,glucagon,β-cell function and insulin resistance among patients with diabetes and metabolic syndrome,so as to discuss the role of pancreatic α cells in pathogenesis of type 2 diabetes mainly caused by insulin resistance and the change of α-cell function after treatment.Methods A total of 60 patients diagnosed with diabetes and metabolic syndrome were selected in Beijing Chaoyang District Diabetes Center from April 2012 to April 2013 and divided with random number table into metformin group (treated with metformin 0.5 g orally thrice a day for 1 year,n =30) and pioglitazone group (treated with pioglitazone 15 mg orally once a day for 1 year,n =30).30 normal healthy people who had physical examination at the Center during the same period were enrolled into the control group,matched in age and gender with the intervention groups.The general condition of the 3 groups,and blood levels of glucose,insulin,and glucagon,insulin sensitivity index (ISI)-Matsuda,homeostasis model assessment of insulin resistance (HOMA-IR),β-cell function index (HOMA-β),1-phase index,2-phase index,and insulin secretion sensitivity index (ISSI) at baseline in the 3 groups and after treatment in the metformin group and the pioglitazone group were measured and calculated.Results Compared with the control group before treatment,the intervention groups as a whole had significantly higher fasting glucagon level [(146.22 ±25.41) pmol/L vs.(21.31 ±7.85) pmol/L,P =0.002] and area under curve (AUC) of glucagon [(469.84 ±13.12) pmol/(L · h) vs.(100.94 ± 7.73) pmol/(L · h),P =0.006].Compared with the results before treatment,the metformin group exhibited significantly reduced fasting glucose [(6.46 ± 1.38) mmol/L vs.(7.54 ± 0.43) mmol/L,P=0.031],fasting insulin [(119.22 ± 69.01) pmol/L vs.(139.38 ±71.13) pmol/L,P =0.042],fasting glucagon [(91.69 ±22.11) pmol/L vs.(142.81 ±24.56) pmol/L,P=0.029],AUC of glucose [(25.19 ± 1.31) mmol/ (L · h) vs.(32.68 ± 1.12) mmol/ (L · h),P =0.043],AUC of insulin [(468.65 ±20.10) pmol/ (L· h) vs.(786.32±21.37) pmol/ (L· h),P=0.017],and AUC of glucagon [(280.60±8.26) pmol/ (L · h) vs.(487.14±14.31) pmol/ (L · h),P=0.032];while the pioglitazone group after treatment also showed significantly decreased fasting glucose [(6.58 ±2.21) mmol/L vs.(7.68±0.59) mmol/L,P=0.028],fastinginsulin [(107.92±17.81) pmol/L vs.(144.66±74.43) pmol/L,P =0.033],fasting glucagon [(76.07 ±20.57) pmol/L vs.(148.34 ±28.94) pmol/L,P=0.025],AUC of glucose [(25.58 ±1.22) mmol/(L·h) vs.(35.07 ±1.38) mmol/(L· h),P=0.038],AUC of insulin [(435.54±19.30) pmol/ (L· h) vs.(854.75 ±20.61) pmol/(L·h),P=0.013],andAUCofglucagon [(223.43 ±5.83) pmol/ (L·h) vs.(458.55 ±12.96) pmol/ (L·h),P =0.026].The before-after-treatment differences were significantly smaller in the metformin group than in the pioglitazone group in terms of fasting insulin [(20.16 ± 2.98) mmol/L vs.(36.74 ± 2.88) mmol/L,P =0.011],fasting glucagon [(51.12 ± 3.67) pmol/L vs.(72.27 ± 4.58) pmol/L,P =0.016],AUC of insulin [(317.67 ±13.45) pmol/(L · h) vs.(419.21 ±15.44) pmol/(L · h),P=0.031] and AUC of glucagon [(206.54±9.66) pmol/(L· h) vs.(235.12±10.29) pmol/(L· h),P=0.046].Conclusions Glucagon in patients with diabetes and metabolic syndrome is higher than that in normal individuals.Metformin and pioglitazone can decrease the level of glucagon in patients with metabolic syndrome and diabetes as well as improve the glucose control,β-cell function and insulin resistance,suggesting improving effect of these two drugs on α-cell function.Pioglitazone manifests a stronger effect than metformin does.

To investigate the effect and mechanism of cytotoxicity by rhein in human renal tubular epithelial HK-2 cells, HgCl2 was choosen as positive control. Cell viability was determined by MTT assay. LDH release assay was used to evaluate cell membrane damage. The activity of caspase-3, and -8 was measured by assay kit. Real-Time qPCR was employed to determine the mRNA expressions of Fas, FasL, FADD, caspase-3 and caspase-8. The protein expressions of Fas, FasL, cytoplasmic cytochrome C, caspase-3, caspase-8, caspase-9 were detected by Western blot. The results demonstrated that rhein inhibited cell viability and increased LDH release in a dose-dependent manner. The activity of caspase-3 and caspase-8 was significantly enhanced by rhein. The mRNA expression of Fas, FasL, FADD, caspase-3, caspase-8 was remarkably up-regulated by rhein. Rhein also elevated protein expressions of Fas, FasL, cytoplasmic cytochrome C, cleaved caspase-3, caspase-8 and reduced expressios of Pro caspase-8. There was no significant difference in caspase-9 expression. These results indicate that rhein has a cytotoxic effect and apoptosis-inducing effect in HK-2 cells. The Fas-dependent pathway is involved in rhein-induced apoptosis.

Objective To study the effects of Pervanadate(Per) on BET-2 cells cell cycle arrest and apoptosis induced by ionizing radiation. Methods The BET-2 cells were radiated with 0, 1, 3, 5, 7, 10 Gy and divided into PTP(protein tyrosine phosphatases)-specific inhibitor Per-treated group and non-pervanadate group(served as a control) and then incubated after ?-ray irradiation. Cell cycle and apoptosis were measured by FCM and Hematoxylin-Eosin staining at different time phases. Results G2/M phase arrest and apoptosis were induced in a pattern of dose-effect and time-course after irradiation in the BET-2 cells. In Per-treated group, G2/M phase arrest increased more notably, and apoptosis decreased markedly. Conclusions These studies suggested that Pervanadate potentialy enhance the ratio and prolong the term of G2/M phase arrest, and facilitate the damaged hematopoietic cells to be repaired, remarkably prevent radiated-cells from apoptosis.