Background and aims:Hepatocellular carcinoma(HCC)is a tumor of high heterogeneity and complexity,which poses significant challenges to effective treatment and patient prognosis because of its immune evasion characteristics.To address these issues,single-cell technology and machine learning methods have emerged as a promising approach to identify genes associated with immune escape in HCC.This study aimed to develop a prognostic risk score model for HCC by identifying intrinsic immune-evasion genes(IIEGs)through single-cell technology and machine learning,providing insights into immune infiltration,enhancing predictive accuracy,and facilitating the development of more effective treatment strategies.Materials and methods:The study utilized data from The Cancer Genome Atlas database to analyze gene expression profiles and clinical data related to intrinsic immune evasion in patients with HCC.Various tools,including the Human Protein Atlas,cBioPortal,single-cell analysis,machine learning,and Kaplan-Meier plot,were used to analyze IIEGs.Functional enrichment analysis was conducted to explore po-tential mechanisms.In addition,the abundance of infiltrating cells in the tumor microenvironment was investigated using single-sample gene set enrichment analysis,CIBERSORT,xCELL,and tumor immu-nophenotype algorithms.The expression of glycosylphosphatidylinositol anchor attachment 1(GPAA1)was examined in the clinical sample of HCC by quantitative real-time polymerase chain reaction,Western blotting,and immunohistochemical staining.Results:Univariate Cox analysis identified 63 IIEGs associated with the prognosis of HCC.Using random forest,least absolute shrinkage and selection operator regression analysis,and support vector machine,a risk score model consisting of six IIEGs(carbamoyl-phosphate synthetase 2,aspartate transcarbamylase,and dihydroorotase(CAD),phosphatidylinositol glycan anchor biosynthesis class U(PIGU),endoplasmic reticulum membrane protein complex subunit 3(EMC3),centrosomal protein 55(CEP55),autophagy-related 10(ATG10),and GPAA1)developed,which was validated using 10 pairs of HCC and adjacent non-cancerous samples.Based on the calculated median risk score,HCC samples were categorized into high-and low-risk groups.The Kaplan-Meier curve analysis showed that the high-risk group had a worse prognosis compared with the low-risk group.Time-dependent receiver operating characteristic analysis demonstrated the accurate predictive capability of the risk score model for HCC prognosis.Furthermore,immune infiltration analysis showed a positive correlation between the risk score model and 40 immune checkpoint genes as well as Th2 cells.Conclusions:A prognostic risk score model was formulated by six IIEG signatures and showed promise in predicting the prognosis of patients diagnosed with HCC.The utilization of the IIEG risk score as a novel prognostic index,together with its significance as a valuable biomarker for immunotherapy in HCC,provides benefit for patients with HCC in determining therapeutic strategies for clinical application.

Objective:To investigate the effect and mechanism of short-chain fatty acids (SCFAs) on the side-effect of tacrolimus on blood glucose.Methods:The C57BL/6 mice were treated with tacrolimus orally (10 mg/kg, tacrolimus group), tacrolimus plus 150 mmol/L sodium butyrate and isovalerate mixed solution (SCFAs group), broad-spectrum antibiotics (antibiotic group), and tacrolimus plus broad-spectrum antibiotics (tac&abx group). After 8 weeks intervention, the fasting blood glucose (FBG), oral glucose tolerance test (OGTT), hemoglobin A1C (HbA1c) were tested as indicators of glucose metabolism, and the gut microbiota, SCFAs concentration in the ileocecal, serum glucagon-like peptide-1 (GLP-1), fasting serum insulin, and GLP-1 expression in intestinal mucosa were performed for intestinal-glucose metabolism mechanism.Results:The FBG and HbA1c were significantly increased in tacrolimus group[(7.31±0.97)mmol/L, (8.34±1.12)%] than control group [(5.23±0.30)mmol/L, (4.32±0.80)%, all P<0.05], which remained normal in antibiotic group [(4.92±0.31)mmol/L, (5.61±0.98)%)], tac&abx group[(5.95±0.37)mmol/L, (4.56±0.26)%] and SCFAs groups [(5.87±0.68)mmol/L, (5.07±1.79)%]. The OGTT in the tacrolimus group showed glucose tolerance impairment, while other groups remained normal. The ileocecal butyric acid and isovaleric acid concentrations in the tacrolimus group were (722.3±262.2) μg/g and (10.0±5.1)μg/g, lower than the control group[ (1 321.3±165.5) μg/g, (19.7±3.6)μg/g, P<0.05]. The above acids in the SCFAs group remained normal as in the control group [(1 375.7±451.6) μg/g, (24.5±11.5)μg/g)]. The fasting serum insulin in the tacrolimus group decreased significantly to (3.2 ± 0.6)mIU/L, compared with control[ (4.4±0.9) mIU/L]and SCFAs groups [(7.0±1.1) mIU/L]. The GLP-1 test indicated a significant decrease in the tacrolimus group[ (4.7±2.9)pg/ml, P<0.05] compared with the SCFAs group and control group [(42.5±19.9) pg/ml, (33.1±9.1) pg/ml]. Conclusions:Tacrolimus affects glucose metabolism through the SCFAs-associated GLP-1 pathway in the intestine, and oral supplementation with mixed SCFAs provides a new insight for the prevention and treatment of tacrolimus-induced hyperglycemia in transplant recipients.

Objective To investigate the correlation between different postoperative serum cortisol cut-off values measured in different periods and the long-term outcomes in patients with Cushing's disease (CD). Methods The clinical data of 102 CD patients undergoing transsphenoidal surgery (TSS) in Peking Union Medical College Hospital from May 1985 to July 2008 were analyzed retrospectively. The differences of long-term outcomes were compared between patients with cortisol levels below 2 μg/dl (2 μg/dl group) and levels between 2 and 5 μg/dl (5 μg/dl group) in the 1postoperative day and 3 and 6 months after surgery. Results The mean follow-up duration was (10.7±1.7) years (range:5-29.1 years). Among these 102 patients,the disease was cured in 74 patients (72.5%) and recurred in 28 patients (27.5%). On the 1postoperative day,there were 63 patients in the 2 μg/dl group,in which 48 patients (76.2%) achieved long-term cure;there were 39 patients in the 5 μg/dl group,in which 26 (66.7%) achieved long-term cure. The difference was not statistically significant (χ=1.097,P=0.295). Three months after TSS,the long-term cure rate was 84.2% (48/57) in the 2 μg/dl group,which was significantly higher than that (65.0%,26/40) in the 5 μg/dl group (χ=4.795,P=0.029). Six months after TSS,the long-term cure rate was 88.7% (47/53) in the 2 μg/dl group,which was significantly higher than that(69.2%,27/39) in the 5 μg/dl group(χ=5.400,P=0.020). Conclusion The serum cortisol level of below 2 μg/dl is more useful than 2-5 μg/dl 3 months and 6 months after surgery in predicting the prognosis of CD patients.
Follow-Up Studies ; Humans ; Hydrocortisone ; blood ; Pituitary ACTH Hypersecretion ; blood ; surgery ; Postoperative Period ; Prognosis ; Recurrence ; Retrospective Studies

BACKGROUND:Exendin can regulate blood glucose, blood lipid and blood pressure, exert anti-inflammation and anti-oxidative stress effects, improve myocardial infarction and heart failure, and protect heart vessels. However, the effect on the apoptosis of cardiac muscle cels after ischemia/reperfusion injury remains unclear. OBJECTIVE:To observe the effects of Exendin-4 pretreatment on the cardiomyocyte apoptosis and the expression of Bcl-2 and Bax in rats with myocardial ischemia/reperfusion injury. METHODS:The myocardial ischemia/reperfusion injury model was established in rats and then received Exendin-4 pretreatment. Ischemia/reperfusion group and sham operation group were set. RESULTS AND CONCLUSION:Immunohistochemical staining, TUNEL and reverse transcriptase-polymerase chain reaction results showed that Bcl-2 protein and mRNA expression levels in the Exendin-4 group were significantly increased (P < 0.05), while Bax protein and mRNA expression levels were significantly decreased compared with ischemia/reperfusion group (P < 0.05). In addition, apoptosis index was more significantly decreased in the Exendin-4 group than in the ischemia/reperfusion group (P < 0.05). Exendin-4 can protect rat heart muscle against ischemia/reperfusion injury and effectively inhibit the apoptosis of cardiomyocytes, and the underlying mechanism is mediated by up-regulating Bcl-2 expression and down-regulating Bax expression.

Objective To explore the influence of intubation-surfactant-extubation (InSurE)therapy combined with bilevel positive airway pressure (BiPAP)in the use time of mechanical ventilation,and to clarify the value of BiPAP in the treatment of respiratory distress syndrome in the preterm infants.Methods Toral 95 preterm infants with respiratory distress syndrome were treated with InSurE therapy during January 2011 to October 2014. Among them,the preterm infants before January 2013 were selected as control group who were treated with InSurE and nasal continuous positive airway pressure (nCPAP).After January 2013, 60 preterm infants were treated with BiPAP,as BiPAP group.The rates of InSurE failure,the need for mechanical ventilation (MV)on the 7th day after InSurE failure, total non-invasive ventilation time, total mechanical ventilation time, atmospheric oxygen therapy time and incidence of clinical complications were compared between two groups.Results ① There were no significant differences in the clinical data of the preterm infants between two groups, such as gender and age.② Although there was no significant difference in the failure rate of InSurE,but the rate of repeated mechanical ventilation during 1 week in BiPAP group was lower than that in control group (P <0.01).③ The Rank sum test result showed that the total time of non invasive ventilation in BiPAP group was longer than that in control group (P <0.01).The total time of invasive mechanical ventilation and oxygen therapy in BiPAP group was lower than that in control group (P < 0.05).④ The incidence of retinopathy of prematurity (ROP)and bronchopulmonary dysplasia (BPD)in BiPAP group was lower than that in control group.Conclusion BiPAP can significantly reduce the use of invasive mechanical ventilation after the failure of InSurE,thereby decreases the oxygen toxicity and barotrauma hazards.