ObjectiveTo investigate the clinical features of patients with acute-on-chronic liver failure （ACLF）， and to construct a risk scoring table that can accurately predict the prognosis of patients in the early stage. MethodsA retrospective analysis was performed for the clinical data of 502 patients with ACLF who were admitted to Tangdu Hospital， Air Force Medical University， from January 1， 2010 to December 31， 2020 （training set）， and the influencing factors for 28-day mortality rate were identified. The 69 ACLF patients who were admitted to Tangdu Hospital， Air Force Medical University， from January 1 to December 31， 2021 were enrolled as the validation set. The independent-samples t test or the Mann-Whitney U test was used for comparison of continuous data between two groups， and the chi-square test or the Fisher’s exact test was used for comparison of categorical data between two groups. A univariate Cox regression analysis was used to obtain the early warning indicators associated with the 28-day prognosis of ACLF patients， and variance inflation factors were used to assess multicollinearity among predictors； a multivariate Cox regression analysis was used to construct a risk model for ACLF prognosis （mortality）. A risk scoring table for ACLF prognosis （mortality） was developed based on regression coefficients （β） from the model equation and weight assignments in the nomogram. Internal validation and comparison were performed for the risk model for ACLF prognosis （mortality）， the scoring table for ACLF prognosis （mortality）， and other scoring models （Child-Turcotte-Pugh ［CTP］ score， Model for End-Stage Liver Disease ［MELD］ score， MELD combined with serum sodium concentration ［MELD-Na］ score， and integrated MELD ［iMELD］ score） in the training set， while external validation and comprehensive evaluation of the scoring table and the other scoring models were performed in the validation set. The Nagelkerke’s R² test and the Hosmer-Lemeshow test were used to assess the degree of fitting of the risk model for ACLF prognosis （mortality）， the scoring table for ACLF prognosis （mortality）， and other scoring models， and fitting curves were plotted. C-index was used to assess the discriminatory ability of the scoring table for ACLF prognosis （mortality） and the other scoring models， and the Z-test was used for comparison of C-index between different models. The decision curve analysis was used to compare the clinical benefits of the scoring table for ACLF prognosis （mortality） and the other scoring models. ResultsThe multivariate Cox regression analysis showed that age （hazard ratio ［HR］=1.027， 95% confidence interval ［CI］： 1.015 — 1.039， P<0.001）， hepatic encephalopathy grade （grade 1： HR=2.928， 95%CI： 1.463 — 5.858， P=0.002； grade 2： HR=3.811， 95%CI： 2.078 — 6.988， P<0.001； grade 3： HR=3.916， 95%CI： 1.917 — 8.001， P<0.001； grade 4： HR=6.966， 95%CI： 4.559 — 10.644， P<0.001）， an increase in total bilirubin （TBil） by ≥17.1 μmol/L per day （HR=1.771， 95%CI： 1.248 — 2.513， P=0.001）， creatinine （HR=1.005， 95%CI： 1.004 — 1.006， P<0.001）， neutrophil count （HR=1.092， 95%CI： 1.060 — 1.126， P<0.001）， and international normalized ratio （HR=1.298， 95%CI： 1.187 — 1.418， P<0.001） were independent risk factors associated with the 28-day mortality rate of ACLF patients， and a risk scoring table was constructed for ACLF prognosis （mortality）. The Nagelkerke’s R² test showed that the risk scoring table for ACLF prognosis （mortality） had an R² value of 0.599 in the training set and 0.722 in the validation set， which were higher than the R² values of CTP， MELD， MELD-Na， and iMELD scores. The Hosmer-Lemeshow test showed that the risk scoring table for ACLF prognosis （mortality） had a P value of 0.280 in the training set and 0.788 in the validation set. The C-index analysis showed that the scoring table had a higher C-index than the other scoring models in the validation set （all P<0.001）， as well as a higher C-index than CTP score in the training set （P<0.001）. The decision curve analysis showed that the risk scoring table for ACLF prognosis （mortality） had higher clinical net benefits than the other scoring models. ConclusionCompared with other scoring models currently used in clinical practice， the novel risk scoring table for ACLF prognosis （mortality） constructed based on the six predictive factors of age， hepatic encephalopathy grade， an increase in TBil by ≥17.1 μmol/L per day， creatinine， neutrophil count， and international normalized ratio has a relatively high value in predicting the 28-day prognosis of ACLF patients.

OBJECTIVES: To establish a prognostic model for primary liver cancer (PLC) using bioinformatics methods. METHODS: Based on the data from 404 patients in the Cancer Genome Atlas (TCGA) database, we constructed a prognostic model integrating the differentially expressed genes, anoikis, and immune-related genes (DAIs) using univariate Cox regression and the LASSO-Cox approach. The predictive ability of the model was evaluated using Kaplan-Meier method and receiver-operating characteristic curves, and a nomogram was developed to facilitate its clinical applications. Gene set enrichment analysis (GSEA) was performed to explore the associated pathways and relationship between the DAIs and the tumor immune microenvironment, and the half-maximal inhibitory concentration (IC₅₀) of liver cancer drugs was calculated using the "pRRophetic" R package. We also detected the expression of SEMA7A in paired tumor and adjacent tissues from liver cancer patients. RESULTS: We constructed and validated a prognostic model based on 7 DAIs (NR4A3, SEMA7A, IL11, AR, BIRC5, EGF, and SPP1), and obtained consistent results in both the TCGA training cohort and GEO validation cohort (GSE14520), where the patients in the low-risk group were characterized by more favorable clinical outcomes and immune status. By integrating this prognostic signature with clinical information, a composite nomogram was generated. Somatic mutation analysis showed that TTN, TP53, and CTNNB1 mutations accounted for the largest proportion of total mutations, and the patients in the low-risk-low-TMB group had higher survival rate. Drug sensitivity analysis revealed differences in sensitivity to chemotherapeutic agents between high- and low-risk groups and between TP53 mutations and non-mutations. In clinical tissue specimens, SEMA7A expression was significantly higher in liver cancer tissues than in the adjacent tissues. CONCLUSIONS We established a new prognostic model based on DAIs for predicting clinical outcomes and therapeutic response of patients with primary liver cancer.
Humans ; Liver Neoplasms/diagnosis* ; Prognosis ; Anoikis ; Nomograms ; Computational Biology ; Tumor Microenvironment ; Semaphorins/metabolism*

At present,some new tongue manifestations with distinct characteristics and high clinical value have not yet gained widespread consensus,which is unfavorable to their research and promotion.This paper representatively explores four types of tongue texture manifestations(liver gall line,bulging vein tongue,concave tip tongue,convex surface tongue)and two types of tongue coating manifestations(cracked tongue coating,white saliva line)among them.It is found that the liver gall line manifests as dark stagnation of various forms on the tongue surface,indicating liver qi stagnation and blood stasis as well as heat-toxin accumulation;the bulging vein tongue is characterized by clearly raised blood vessels on the tongue surface,suggesting blood stasis due to yang deficiency or cold congelation;the concave tip tongue is characterized by a depressed tip,indicating an abnormal tongue frenulum or malnutrition of heart,lung,and kidney;the convex surface tongue features a broad and thick tongue body with an overall"convex"shape,which is observed in healthy individuals or suggests qi movement disorder and spleen and stomach stagnation;the cracked tongue coating is characterized by cracks only on the tongue coating,indicating qi deficiency with disordered fluid or yin consumption due to intense heat;the white saliva line presents as strip-like white foam bands on the tongue margin,suggesting liver qi stagnation and dampness stagnancy due to spleen deficiency.No consensus exists regarding issues such as naming and description,reflecting the necessity of expanding and standardizing the theory of tongue diagnosis.This paper aims to emphasize the value and standardization of such new tongue manifestations,enrich the theoretical system of tongue diagnosis in traditional Chinese medicine,and provide novel insights and reference basis for clinical syndrome differentiation.

This study investigates the feasibility of the VP8*protein as a subunit vaccine target for porcine rotavirus A(PoRVA),a major causative agent of diarrhea in piglets.The VP8* genes of PoRVA P[13]and P[23]genotype strains were amplified by RT-PCR.These genes were then liga-ted into the pET-28a(+)vector,yielding recombinant plasmids pET-28a-XJWF1-VP8*-P[23]and pET-28a-ShXYW13-VP8*-P[13].These plasmids were subsequently transformed into BL21(DE3)competent cells.The VP8*protein,induced by IPTG,was purified using affinity chroma-tography,and its expression and purification were verified by SDS-PAGE and Western blot.The purified VP8* protein was used to immunize mice,and serum samples were collected after three immunizations.Cross-neutralization assays were conducted to evaluate the ability of the VP8*protein immune serum to neutralize different genotype strains.The results demonstrated the ex-pression of soluble VP8*protein,with SDS-PAGE and Western blot analyses showing that the purified VP8*protein existed in both monomeric(27 kDa)and homodimeric(54 kDa)forms.ELISA results indicated that high levels of antibodies were produced in mice immunized with VP 8*-P[13]and VP8*-P[23]after three immunizations.Serum cross-neutralization assays revealed that the neutralizing titers of PoRVA VP8*-P[13]and VP8*-P[23]immune sera against homol-ogous genotype strains ranged from 1∶4 800 to 1∶19 200,significantly higher than those against heterologous genotype strains(1∶1 200).This suggests that the VP8*protein of different geno-type strains exhibits both antigenic conservation and distinct variability.The data obtained in this study provide a solid foundation for further exploration of the antigenic structure of the PoRVA VP8* protein and the development of novel subunit vaccines.

This study investigates the feasibility of the VP8*protein as a subunit vaccine target for porcine rotavirus A(PoRVA),a major causative agent of diarrhea in piglets.The VP8* genes of PoRVA P[13]and P[23]genotype strains were amplified by RT-PCR.These genes were then liga-ted into the pET-28a(+)vector,yielding recombinant plasmids pET-28a-XJWF1-VP8*-P[23]and pET-28a-ShXYW13-VP8*-P[13].These plasmids were subsequently transformed into BL21(DE3)competent cells.The VP8*protein,induced by IPTG,was purified using affinity chroma-tography,and its expression and purification were verified by SDS-PAGE and Western blot.The purified VP8* protein was used to immunize mice,and serum samples were collected after three immunizations.Cross-neutralization assays were conducted to evaluate the ability of the VP8*protein immune serum to neutralize different genotype strains.The results demonstrated the ex-pression of soluble VP8*protein,with SDS-PAGE and Western blot analyses showing that the purified VP8*protein existed in both monomeric(27 kDa)and homodimeric(54 kDa)forms.ELISA results indicated that high levels of antibodies were produced in mice immunized with VP 8*-P[13]and VP8*-P[23]after three immunizations.Serum cross-neutralization assays revealed that the neutralizing titers of PoRVA VP8*-P[13]and VP8*-P[23]immune sera against homol-ogous genotype strains ranged from 1∶4 800 to 1∶19 200,significantly higher than those against heterologous genotype strains(1∶1 200).This suggests that the VP8*protein of different geno-type strains exhibits both antigenic conservation and distinct variability.The data obtained in this study provide a solid foundation for further exploration of the antigenic structure of the PoRVA VP8* protein and the development of novel subunit vaccines.

At present,some new tongue manifestations with distinct characteristics and high clinical value have not yet gained widespread consensus,which is unfavorable to their research and promotion.This paper representatively explores four types of tongue texture manifestations(liver gall line,bulging vein tongue,concave tip tongue,convex surface tongue)and two types of tongue coating manifestations(cracked tongue coating,white saliva line)among them.It is found that the liver gall line manifests as dark stagnation of various forms on the tongue surface,indicating liver qi stagnation and blood stasis as well as heat-toxin accumulation;the bulging vein tongue is characterized by clearly raised blood vessels on the tongue surface,suggesting blood stasis due to yang deficiency or cold congelation;the concave tip tongue is characterized by a depressed tip,indicating an abnormal tongue frenulum or malnutrition of heart,lung,and kidney;the convex surface tongue features a broad and thick tongue body with an overall"convex"shape,which is observed in healthy individuals or suggests qi movement disorder and spleen and stomach stagnation;the cracked tongue coating is characterized by cracks only on the tongue coating,indicating qi deficiency with disordered fluid or yin consumption due to intense heat;the white saliva line presents as strip-like white foam bands on the tongue margin,suggesting liver qi stagnation and dampness stagnancy due to spleen deficiency.No consensus exists regarding issues such as naming and description,reflecting the necessity of expanding and standardizing the theory of tongue diagnosis.This paper aims to emphasize the value and standardization of such new tongue manifestations,enrich the theoretical system of tongue diagnosis in traditional Chinese medicine,and provide novel insights and reference basis for clinical syndrome differentiation.

ObjectiveTo investigate the clinical features of patients with acute-on-chronic liver failure （ACLF） and bacterial infection and early warning indicators associated with multidrug-resistant infections. MethodsA retrospective analysis was performed for 130 patients with ACLF and bacterial infection who attended The Second Affiliated Hospital of Air Force Medical University from January 1， 2010 to December 31， 2021， and according to the drug susceptibility results， the patients were divided into multidrug-resistant （MDR） bacterial infection group with 80 patients and non-MDR bacterial infection group with 50 patients. General information and laboratory examination results were compared between the two groups to screen for the early warning indicators associated with MDR bacterial infection. The Student’s t-test was used for comparison of normally distributed continuous data with homogeneity of variance between two groups， and the Mann-Whitney U test was used for comparison of non-normally distributed continuous data or continuous data with heterogeneity of variance between two groups； the chi-square test or the Fisher’s exact test was used for comparison of categorical data between two groups. The binary logistic regression analysis and the receiver operating characteristic （ROC） curve were used to assess the predictive value of early warning indicators. ResultsAmong the 130 patients with ACLF and bacterial infection， sputum （27.7%） was the most common specimen for detection， followed by blood （24.6%）， urine （18.5%）， and ascites （17.7%）. Bacterial infections were dominated by Gram-negative bacteria （58.5%）. Of all bacteria， Escherichia coli （18.5%）， Klebsiella pneumoniae （14.6%）， and Enterococcus faecium （13.8%） were the most common pathogens. Gram-positive bacteria had a high resistance rate to the antibacterial drugs such as erythromycin （72.2%）， penicillin （57.4%）， ampicillin （55.6%）， and ciprofloxacin （53.7%）， while Gram-negative bacteria had a high resistance rate to the antibacterial drugs such as ampicillin （73.3%）， cefazolin （50.0%）， and cefepime （47.4%）. The patients with ACLF and bacterial infection had a relatively high rate of MDR bacterial infection （61.5%）. Comparison of clinical data between the two groups showed that compared with the patients with non-MDR bacterial infection， the patients with MDR bacterial infection had significantly higher levels of alanine aminotransferase （Z=2.089， P=0.037）， aspartate aminotransferase （Z=2.063， P=0.039）， white blood cell count （Z=2.207， P=0.027）， and monocyte count （Z=4.413， P<0.001）. The binary logistic regression analysis showed that monocyte count was an independent risk factor for MDR bacterial infection （odds ratio=7.120， 95% confidence interval ［CI］： 2.478‍ ‍—‍ ‍20.456，P<0.001） and had an area under the ROC curve of 0.686 （95%CI： 0.597‍ ‍—‍ ‍0.776） in predicting ACLF with MDR bacterial infection（P<0.001）， with the optimal cut-off value of 0.50×10⁹/L， a sensitivity of 0.725， and a specificity of 0.400. ConclusionACLF combined with bacterial infections is mainly caused by Gram-negative bacteria， with the common pathogens of Escherichia coli and Klebsiella pneumoniae and a relatively high MDR rate in clinical practice. An increase in monocyte count can be used as an early warning indicator to distinguish MDR bacterial infection from non-MDR bacterial infection.

Objective:To investigate the correlation between fibrinogen/albumin ratio (FAR) and overall burden of cerebral small vessel disease (CSVD).Methods:Patients with CSVD who visited the Department of Neurology, Zhengzhou Central Hospital Affiliated to Zhengzhou University from May 2023 to May 2024 were included retrospectively. All patients underwent MRI to evaluate the total CSVD burden score (range: 0-4) by cerebral microbleeds, white matter hyperintensities, lacunar and enlarged perivascular spaces. 0-1 point was the mild burden group, and 2-4 points were the moderate-to-severe burden group. FAR was calculated as serum fibrinogen (g/L)/albumin (g/L)×100%. Spearman correlation analysis and multivariate logistic regression analysis were used to determine the relationship between FAR and total CSVD burden. Results:A total of 261 patients with CSVD were enrolled, including 145 males (55.6%), aged 64.08±8.74 years. There were 110 patients (42.1%) in the CSVD mild burden group and 151 (57.9%) in the moderate-to-severe burden group. Spearman correlation analysis showed that there was a significant positive correlation between FAR and total CSVD burden scores ( r=0.385, P<0.001). Univariate analysis showed that the proportion of patients with diabetes, history of stroke or transient ischemic attack, as well as age, FAR, homocysteine and fibrinogen in the CSVD moderate-to-severe burden group were significantly higher than those in the mild burden group, while serum albumin and high-density lipoprotein cholesterol were significantly lower in the mild burden group (all P<0.05). Multivariate logistic regression analysis showed that higher FAR (odds ratio 1.433, 95% confidence interval 1.201-1.710; P<0.001) and homocysteine (odds ratio 1.093, 95% confidence interval 1.026-1.164; P=0.005) were significantly independently associated with the moderate-to-severe total CSVD burden. Conclusion:The level of FAR is closely associated with the severity of the total CSVD burden in patients with CSVD.

Objective:To investigate the expression of interleukin-10 (IL-10) and chemokine receptor 7 (CXCR7) in papillary thyroid carcinoma (PTC) tissues and their correlation with invasion and metastasis.Methods:According to the inclusion and exclusion criteria, 68 cases of cancer tissues (cancer tissue group) and para-cancer tissues (para-cancer tissue group) collected from PTC patients in Wenzhou Hospital of Traditional Chinese Medicine from Jun. 2015 to Jun. 2018 were selected. Meanwhile, pathological specimens from 32 cases of thyroid follicular adenoma patients (thyroid follicular adenoma group) undergoing thyroidectomy during the same period were also enrolled. Immunohistochemical SP staining was used to observe the expression of IL-10 and CXCR7 in different tissues, and the correlation between the expression of IL-10 and CXCR7 and the pathological characteristics of tumor was analyzed.Results:The positive expression rates of IL-10 and CXCR7 in cancer tissues were significantly higher than those in para-cancer tissues and thyroid follicular adenoma group (88.24% vs. 14.71% vs. 0.00%, 75.00% vs. 32.35% vs. 15.63%; P<0.05) . Among 68 PCT patients, 38 (55.88%) had high expression of IL-10, 30 (44.12%) had low expression of IL-10, 32 (47.06%) had high expression of CXCR7, and 36 (52.94%) had low expression of CXCR7. IL-10 expression was positively correlated with tumor multifocal, TNM stage, central lymphatic metastasis, central + cervical lymphatic metastasis, invasion degree and tumor maximum diameter ( r=0.486, 0.509, 0.468, 0.422, 0.629, 0.674, P<0.05) . CXCR7 expression was positively correlated with tumor multifocal, TNM staging, central lymphatic metastasis, central + cervical lymphatic metastasis, and infiltration (r=0.425, 0.563, 0.490, 0.287, 0.481, P<0.05) . Conclusions:IL-10 and CXCR7 are highly expressed in cancer tissues of PTC patients, among which IL-10 expression level is closely related to tumor differentiation degree, TNM stage, cervical lymphatic metastasis, infiltration degree and tumor maximum diameter. CXCR7 is related to TNM stage, cervical lymphatic metastasis and infiltration degree, both of which may be involved in the occurrence and development of tumor.

Objective To explore the clinicopathological characteristics,recurrence,metastasis and survival of triple negative breast cancer (TNBC),and to analyze the correlation factors affecting the prognosis.Methods Data of 378 breast cancer patients treated from Jan.2008 to Dec.2011 were retrospectively analyzed.According to immunohistochemical staining of estrogen of receptor (ER),progesterone receptor (PR) and human epidermal growth factor receptor 2 (HER2),they were divided into TNBC group (61 cases) and non-triple negative breast cancer group (non-TNBC,317 cases).The two groups were compared in terms of clinicopathological characteristics,prognosis and survival.Results Patients in TNBC group had significant differences in the following aspects:ratio of patients with newly diagnosed age ＜35 years old,patients with family history of breast cancer,the maximum diameter of tumor more than 5 cm,positive preoperative axillary lymph node status,tumor in clinical stage m,histological grade of tumor in level Ⅲ,Ki67 overexpression and P53 overexpression,while there was no statistical difference in the aspects of menstrual status,pathological type or surgical method between them.The local recurrence and distant metastasis rate were obviously higher in TNBC group than in non-TNBC group.5-year diseasefree survival (DFS) and 5-year overall survival (OS) were significantly lower in TNBC group than in non-TNBC group.Univariate analysis showed that factors related to 5-year DFS in TNBC group were:age,the maximum diameter of tumor,the preoperative axillary lymph node status,clinical staging of tumor,and P53 overexpression.The maximum diameter of tumor,the preoperative axillary lymph node status and clinical staging of tumor were recognized as the influence factors of 5-year OS.Independent factors affecting 5-year DFS in TNBC group were:the maximum diameter of tumor,and the preoperative axillary lymph node status.The maximum diameter of tumor and the preoperative axillary lymph node status were the independent factors influencing 5-year OS.Conclusions The clinicopathological characteristics of TNBC include:younger onset age,family clustering of breast cancer,the larger maximum of tumor diameter,larger portion of positive preoperative axillary lymph node,later clinical staging of tumor,higher histological grade of tumor,easier local recurrence and distant metastasis,lower 5-year DFS and 5-year OS.The factors of age,the maximum diameter of tumor,the preoperative axillary lymph node status,clinical staging of tumor,P53 overexpression especially the maximum diameter of tumor and the preoperative axillary lymph node status play the important clinical roles in judging the prognosis of TNBC.