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.

ObjectiveTo investigate the protective effect of Rehmanniae Radix iridoid glycosides （RIG） on the kidney tissue of streptozotocin （STZ）-induced diabetic mice and explore the underlying mechanism. MethodsTwelve of 72 male C57BL/6J mice were randomly selected as the normal group， and the remaining 60 mice were fed with a high-fat diet for six weeks combined with injection of 60 mg·kg^-1 STZ for 4 days to model type 2 diabetes mellitus. The successfully modeled mice were randomized into model， metformin （250 mg·kg^-1）， catalpol （100 mg·kg^-1）， low-dose RIG （RIG-L， 200 mg·kg^-1） and high-dose RIG （RIG-H， 400 mg·kg^-1） groups （n=11）. Mice in each group were administrated with corresponding drugs， while those in the normal group and model group were administrated with the same dose of distilled water by gavage once a day. After 8 weeks of intervention， an oral glucose tolerance test （OGTT） was performed， and the area under the curve （AUC） was calculated. After mice were sacrificed， both kidneys were collected. The body weight， kidney weight， and fasting blood glucose （FBG） were measured. Biochemical assays were performed to measure the serum levels of triglycerides （TG）， total cholesterol （TC）， serum creatinine （SCr）， and blood urea nitrogen （BUN）. Enzyme-linked immunosorbent assay （ELISA） was employed to determine the serum level of fasting insulin （FINS）， and the insulin sensitivity index （ISI） and homeostatic model assessment for insulin resistance （HOMA-IR） were calculated. The pathological changes in kidneys of mice were observed by hematoxylin-eosin staining and Masson staining. The immunohistochemical method （IHC） was employed to assess the expression of interleukin-1 （IL-1）， interleukin-6 （IL-6）， tumor necrosis factor-α（TNF-α）， transforming growth factor-β₁ （TGF-β₁）， and collagen-3 （ColⅢ） in the kidney tissue. The protein levels of TGF-β₁， cell signal transduction molecule 3 （Smad3）， matrix metalloproteinase-9 （MMP-9）， and ColⅢ in kidneys of mice were determined by Western blot. ResultsCompared with the normal group， the model group showcased decreased body weight and ISI （P<0.01）， increased kidney weight， FBG， AUC， FINS， HOMA-IR， TC， TG， SCr， and BUN （P<0.01）， glomerular hypertrophy， capsular space narrowing， and collagen deposition in the kidney， up-regulated protein levels of IL-1， IL-6， TNF-α， TGF-β₁， ColⅢ， and Smad3 （P<0.01）， and down-regulated protein level of MMP-9 （P<0.01） in the kidney tissue. Compared with the model group， the treatment groups had no significant difference in the body weight and decreased kidney weight （P<0.05， P<0.01）. The FBG level declined in the RIG-H group after treatment for 4-8 weeks and in the metformin， catalpol， and RIG-L groups after treatment for 6-8 weeks （P<0.01）. The AUC in the RIG-L， RIG-H， and metformin groups decreased （P<0.05， P<0.01）. The levels of TC， SCr， and BUN in the serum of mice in each treatment group became lowered （P<0.05， P<0.01）. The level of TG declined in the RIG-L， RIG-H， and metformin groups （P<0.05， P<0.01）. The serum level of FINS declined in the catalpol， RIG-L， and metformin groups （P<0.01）. Compared with the model group， the treatment groups showed decreased HOMA-IR （P<0.01）， increased ISI （P<0.01）， alleviated pathological changes in the kidney tissue， and down-regulated expression of IL-1 and TGF-β₁. In addition， the protein levels of IL-6， TNF-α， and ColⅢ in the RIG-H and metformin groups and IL-6 and TNF-α in the RIG-L group were down-regulated （P<0.05， P<0.01）， and the protein levels of IL-6， TNF-α， and ColⅢ in the catalpol group and ColⅢ in the RIG-L group showed a decreasing trend without statistical difference. The protein levels of TGF-β₁， Smad3， and ColⅢ in the RIG-H and metformin groups were down-regulated （P<0.01）. Compared with that in the model group， the protein level of MMP-9 was up-regulated in each treatment group （P<0.01）. ConclusionRIG can improve the renal structure and function of diabetic mice by regulating the TGF-β₁/Smads signaling pathway.

Peanut, a major economic and oil crop known for the high protein and oil content, is extensively cultivated in China. Peanut plants have the ability to form nodules with rhizobia, where the nitrogenase converts atmospheric nitrogen into ammonia nitrogen that can be utilized by the plants. Analysis of nodule fixation is of positive significance for avoiding overapplication of chemical fertilizer and developing sustainable agriculture. In this study, AhNIGT1.2, a member of the NIGT family predominantly expressed in peanut nodules, was identified by bioinformatics analysis. Subsequent spatiotemporal expression analysis revealed that AhNIGT1.2 was highly expressed in nodules and showed significant responses to high nitrogen, low nitrogen, high phosphorus, low phosphorus, and rhizobia treatments. Histochemical staining indicated that the gene was primarily expressed in developing nodules and at the connection region between mature nodules and peanut roots. The fusion protein AhNIGT1.2-GFP was located in the nucleus of tobacco epidermal cells. The AhNIGT1.2-OE significantly increased the number of peanut nodules, while AhNIGT1.2-RNAi reduced the number of nodules, which suggested a positive regulatory role of AhNIGT1.2 in peanut nodulation. The AhNIGT1.2-OE in roots down-regulated the expression levels of NRT1.2, NRT2.4, NLP1, and NLP7, which indicated that AhNIGT1.2 influenced peanut nodulation by modulating nitrate transport and the expression of NLP genes. The transcriptome analysis of AhNIGT1.2-OE and control roots revealed that overexpressing AhNIGT1.2 significantly enriched the differentially expressed genes associated with nitrate response, nodulation factor pathway, enzymes for triterpene biosynthesis, and carotenoid biosynthesis. These findings suggest that AhNIGT1.2 play a key role in peanut nodulation by regulating nitrate transport and response and other related pathways. This study gives insights into the molecular mechanisms of nitrogen and phosphorus in regulating legume nodulation and nitrogen fixation, and sheds light on the development of legume crops that can efficiently fix nitrogen in high nitrogen environments.
Arachis/physiology* ; Nitrates/metabolism* ; Plant Proteins/physiology* ; Transcription Factors/metabolism* ; Plant Root Nodulation/physiology* ; Gene Expression Regulation, Plant ; Root Nodules, Plant/metabolism* ; Nitrogen Fixation

Objective To analyze the expression levels of matrix metalloproteinase-10(MMP-10)and Toll-like receptor 2(TLR2)in serum of patients underwent decompression surgery(DC)for severe traumatic brain injury(sTBI),and to explore their relationship with disease outcome.Methods From April 2021 to April 2024,sTBI patients(n=94)who received DC treatment in a single center were collected as the observation group.Another 90 healthy volunteers who underwent physical examinations at our hospital were selected as the control group.Six months after surgery,sTBI patients were assigned into the good group(n=53)and the adverse group(n=41)according to the Glasgow Outcome Scale(GOS).Data was collected from each group and their differences were compared.Enzyme linked immunosorbent assay(ELISA)was used to measure serum levels of MMP-10 and TLR2.Spearman method was used to analyze the correlation between MMP-10,TLR2 levels and disease outcomes.Logistic regression model used to analyze influencing factors of disease outcomes in sTBI patients after DC.The receiver operating characteristic(ROC)curve was applied to evaluate the predictive value of serum MMP-10 and TLR2 levels for disease outcome in sTBI patients after DC.Results Compared with the control group,the expression levels of serum MMP-10 and TLR2 were prominently higher in the observation group(P＜0.05).Compared with the good group,the proportions of sTBI patients with cerebral herniation,multiple brain contusions and lacerations,and serum levels of MMP-10 and TLR2 were significantly higher in the adverse group,while Glasgow Coma Scale(GCS)score was significantly lower(P＜0.05).Serum levels of MMP-10 and TLR2 in sTBI patients were positively correlated with poor prognosis after DC(P＜0.05).Elevated levels of serum MMP-10 and TLR2,and the increased proportions of patients with cerebral herniation and multiple brain contusions were risk factors affecting the disease outcome after DC in sTBI patients,while elevated GCS score was a protective factor(P＜0.05).The area under the curve(AUC)for predicting disease outcome in sTBI patients after DC using serum MMP-10 and TLR2 alone and in combination was 0.839(95％CI:0.749-0.907),0.847(95％CI:0.758-0.913)and 0.925(95％CI:0.852-0.969),respectively.The combined detection was superior to the individual detections(Zcombination-MMP-10=2.199,Zcombination-TLR2=2.377,both P＜0.05).Conclusion The expression levels of serum MMP-10 and TLR2 in sTBI patients are significantly elevated,and both are prominently correlated with disease outcome after DC.

ObjectiveTo explore the changes in volatile components， total polysaccharides， enzyme activity， and chromaticity value of Myristicae Semen Koji（MSK） during the fermentation process， and conduct correlation analysis. MethodsBased on gas chromatography-mass spectrometry（GC-MS）， the changes of volatile components in MSK at different fermentation times were identified. The phenol sulfuric acid method， dinitrosalicylic acid method（DNS）， and carboxymethyl cellulose sodium salt method（CMC-Na） were used to investigate the total polysaccharide content， amylase activity， and cellulase activity during the fermentation process. Visual analysis technology was used to explore the changes in chromaticity values， revealing the fermentation process of MSK and the dynamic changes of various measurement indicators， partial least squares-discriminant analysis（PLS-DA） was used to explore the differential compounds of MSK at different fermentation degrees， and Pearson correlation analysis was used to explore the correlation between volatile components of MSK and total polysaccharides， enzyme activity， and chromaticity values. ResultsA total of 60 volatile compounds were identified from MSK， the relative contents of components such as （+）-α-pinene， β-phellandrene， β-pinene， （+）-limonene， and p-cymene obviously increased， while the relative contents of components such as safrole， methyl isoeugenol， methyleugenol， myristicin， and elemicin significantly decreased. During the fermentation process， the total polysaccharide content showed an upward trend， while the activities of amylase and cellulase showed an initial increase followed by a decrease， and reached their maximum value at 40 h. the overall brightness（L^*） and total color difference（ΔE^*） gradually increased， while the changes in red-green value（a^*） and yellow-blue value（b^*） were not obvious. PLS-DA results showed that MSK could be clearly distinguished at different fermentation times， and 13 differential biomarkers were screened out. Pearson correlation analysis results showed that the contents of α-terpinene， β-phellandrene， methyleugenol， β-cubebene and myristic acid had an obvious correlation with chromaticity values. ConclusionAfter fermentation， the volatile components， total polysaccharides， amylase activity， and cellulase activity of MSK undergo significant changes， and there is a clear correlation between them and chromaticity values， which reveals the dynamic changes in the fermentation process and related indicators of MSK， laying a foundation for the quality control.

Abstract Tumor immune escape represents a pivotal determinant of cancer immunotherapy failure. It's mechanistically linked to immunosuppressive tumor microenvironment(TME). The TME comprises tumor cells, immune cells, stromal components and extracellular matrix. These components interact synergistically to suppress antitumor immunity through multiple pathways, thereby promoting immune evasion. As crucial chromosomal stability regulators, centromere proteins(CENPs) remodel the TME via multifaceted mechanisms to potentiate immune evasion. This review synthesizes current knowledge on CENPs' role in tumor immune evasion, offering novel insights for cancer diagnostics and immunotherapy.

Objective:To study the current prevalence and temporal patterns of Keshan disease in Gansu Province, thereby establishing a scientific foundation for development of targeted prevention and control strategies.Methods:From March 2019 to October 2021, monitoring of Keshan disease was conducted in all affected townships (towns) across 28 counties (districts, cities) in Gansu Province. Demographic data were collected from affected villages, and suspected chronic Keshan disease cases were identified through village clinics, township health centers, county hospitals, and other medical institutions. These suspected cases were diagnostically verified according to the "Diagnosis of Keshan Disease" (WS/T 210-2011). Current chronic Keshan disease cases underwent re-examination and follow-up monitoring.Results:There were a total of 254 endemic townships (towns) and 2 611 endemic villages across the province, with a permanent resident population of 3.667 1 million in the affected areas. New cases of chronic Keshan disease continued to emerge throughout the study period, totaling 6 cases. From 2020 to 2021, among the total number of chronic Keshan disease cases, 88.92% (923/1 038) remained stable in condition, while mortality cases accounted for 9.54% (99/1 038). Over the past three years, there were 886 males and 687 females with current chronic Keshan disease, mainly aged between 50 and 60 years old, accounting for 38.78% (610/1 573).Conclusions:While Keshan disease in Gansu Province maintains a sustained level of elimination, new chronic cases continue to emerge. It is essential to enhance disease surveillance efforts, and effectively consolidate the achievements in Keshan disease prevention and control measures.

Objective:To study the current prevalence and temporal patterns of Keshan disease in Gansu Province, thereby establishing a scientific foundation for development of targeted prevention and control strategies.Methods:From March 2019 to October 2021, monitoring of Keshan disease was conducted in all affected townships (towns) across 28 counties (districts, cities) in Gansu Province. Demographic data were collected from affected villages, and suspected chronic Keshan disease cases were identified through village clinics, township health centers, county hospitals, and other medical institutions. These suspected cases were diagnostically verified according to the "Diagnosis of Keshan Disease" (WS/T 210-2011). Current chronic Keshan disease cases underwent re-examination and follow-up monitoring.Results:There were a total of 254 endemic townships (towns) and 2 611 endemic villages across the province, with a permanent resident population of 3.667 1 million in the affected areas. New cases of chronic Keshan disease continued to emerge throughout the study period, totaling 6 cases. From 2020 to 2021, among the total number of chronic Keshan disease cases, 88.92% (923/1 038) remained stable in condition, while mortality cases accounted for 9.54% (99/1 038). Over the past three years, there were 886 males and 687 females with current chronic Keshan disease, mainly aged between 50 and 60 years old, accounting for 38.78% (610/1 573).Conclusions:While Keshan disease in Gansu Province maintains a sustained level of elimination, new chronic cases continue to emerge. It is essential to enhance disease surveillance efforts, and effectively consolidate the achievements in Keshan disease prevention and control measures.

Objective To analyze the expression levels of matrix metalloproteinase-10(MMP-10)and Toll-like receptor 2(TLR2)in serum of patients underwent decompression surgery(DC)for severe traumatic brain injury(sTBI),and to explore their relationship with disease outcome.Methods From April 2021 to April 2024,sTBI patients(n=94)who received DC treatment in a single center were collected as the observation group.Another 90 healthy volunteers who underwent physical examinations at our hospital were selected as the control group.Six months after surgery,sTBI patients were assigned into the good group(n=53)and the adverse group(n=41)according to the Glasgow Outcome Scale(GOS).Data was collected from each group and their differences were compared.Enzyme linked immunosorbent assay(ELISA)was used to measure serum levels of MMP-10 and TLR2.Spearman method was used to analyze the correlation between MMP-10,TLR2 levels and disease outcomes.Logistic regression model used to analyze influencing factors of disease outcomes in sTBI patients after DC.The receiver operating characteristic(ROC)curve was applied to evaluate the predictive value of serum MMP-10 and TLR2 levels for disease outcome in sTBI patients after DC.Results Compared with the control group,the expression levels of serum MMP-10 and TLR2 were prominently higher in the observation group(P＜0.05).Compared with the good group,the proportions of sTBI patients with cerebral herniation,multiple brain contusions and lacerations,and serum levels of MMP-10 and TLR2 were significantly higher in the adverse group,while Glasgow Coma Scale(GCS)score was significantly lower(P＜0.05).Serum levels of MMP-10 and TLR2 in sTBI patients were positively correlated with poor prognosis after DC(P＜0.05).Elevated levels of serum MMP-10 and TLR2,and the increased proportions of patients with cerebral herniation and multiple brain contusions were risk factors affecting the disease outcome after DC in sTBI patients,while elevated GCS score was a protective factor(P＜0.05).The area under the curve(AUC)for predicting disease outcome in sTBI patients after DC using serum MMP-10 and TLR2 alone and in combination was 0.839(95％CI:0.749-0.907),0.847(95％CI:0.758-0.913)and 0.925(95％CI:0.852-0.969),respectively.The combined detection was superior to the individual detections(Zcombination-MMP-10=2.199,Zcombination-TLR2=2.377,both P＜0.05).Conclusion The expression levels of serum MMP-10 and TLR2 in sTBI patients are significantly elevated,and both are prominently correlated with disease outcome after DC.

Objective:To study the correlation between serum N-terminal pro-B-type natriuretic peptide (NT-proBNP) level and echocardiographic parameters in patients with chronic Keshan disease, providing reference for diagnosis and prognosis evaluation of chronic Keshan disease.Methods:Ninety-nine patients with chronic Keshan disease who received standardized treatment at Jingchuan County People's Hospital in Pingliang City, Gansu Province from January to December 2020 were selected. Among them, 16 patients were classified as cardiac function grade Ⅱ according to New York Heart Association (NYHA), 69 as grade Ⅲ and 14 as grade Ⅳ. The patients underwent echocardiography and their serum NT-proBNP level was measured using fluorescence immunochromatography. The differences in serum NT-proBNP levels among patients with different cardiac function grades were compared, and the correlation between cardiac function grades, serum NT-proBNP level and echocardiographic parameters was analyzed.Results:The serum NT-proBNP levels in patients with cardiac function grades Ⅱ, Ⅲ, and Ⅳ were (1 107.26 ± 268.03), (2 125.98 ± 293.02), and (8 268.59 ± 2 659.50) pg/ml, respectively. The differences among the three groups were statistically significant ( F = 13.94, P < 0.001). The serum NT-proBNP level was positively correlated with cardiac function grades ( r = 0.44, P < 0.001), left ventricular end-diastolic diameter, left ventricular end-systolic diameter, and left atrial diameter ( r = 0.45, 0.52, 0.38, P < 0.001), and negatively correlated with fractional shortening and left ventricular ejection fraction ( r = - 0.39, - 0.46, P < 0.001). Conclusions:The serum NT-proBNP level in patients with chronic Keshan disease with different cardiac function grades is different, and is positively correlated with echocardiographic parameters reflecting the degree of cardiac structural and functional impairment. The NT-proBNP level may become an early diagnostic, grading, and prognostic indicator for chronic Keshan disease.