ObjectiveTo investigate the role of DEAD-box helicase 3 X-linked （DDX3X） in immune-mediated liver injury （ILI）， and to clarify its mechanism by regulating endoplasmic reticulum stress （ERS）-dependent apoptotic pathway and its association with the clinical progression of hepatitis B. MethodsMice were given injection of concanavalin A （ConA） via the caudal vein to establish a model of ILI， PBS （control group） and different concentrations of ConA were injected into the tail vein of hepatocyte-specific DDX3X-knockout mice （DDX3X^ΔHep and DDX3X-flox mice （DDX3X^fl/fl）， respectively.. The log-rank survival analysis， measurement of the serum levels of aspartate aminotransferase （AST） and alanine aminotransferase （ALT）， and HE staining of liver tissue were performed to assess liver injury， and qRT-PCR and Western Blot were used to measure the mRNA and protein expression levels of glucose-regulated protein 78 （GRP78）， CCAAT/enhancer-binding protein homologous protein （CHOP）， and DDX3X in liver tissue. Intraperitoneal injection of 4-phenylbutyric acid （4-PBA， 100 mg/kg） was performed to inhibit ERS. Serum samples （n=30） and liver tissue samples （n=6） were collected from healthy controls， chronic hepatitis B （CHB） patients， and hepatitis B virus-associated liver failure （HBV-LF） patients； ELISA was used to measure the serum level of DDX3X， and qRT-PCR/Western Blot was used to analyze the expression of targets in liver tissue. A one-way analysis of variance was used for comparison of continuous data between multiple groups， and the least significant difference t-test was used for further comparison between two groups. ResultsCompared with the control group of mice， the expression of DDX3X in the liver of mice induced by ConA was significantly increased after liver injury （P<0.05）， and hepatocyte-specific DDX3X knockout increased the 72-hour survival rate of mice by 55% （compared with 20% in the DDX3X^fl/fl group）， with significant reductions in the serum levels of ALT and AST （P<0.000 1） and the expression levels of the ERS markers GRP78 and CHOP （P<0.05）. After ERS was inhibited by 4-PBA， there was alleviation of liver injury （with reductions in ALT and AST， P <0.001） and a reduction in DDX3X expression （P<0.01）. The analysis of clinical samples showed that the mRNA and protein expression levels of liver DDX3X in CHB patients and HBV-LF patients were significantly higher than those in healthy controls （all P<0.01）， and there was a significant increase in the serum level of DDX3X in HBV-LF patients （P<0.000 1）. ConclusionDDX3X exacerbates ILI by regulating the ERS-dependent apoptotic pathway （GRP78/CHOP）， and its expression is associated with the progression of hepatitis B. Therefore， it can be used as a potential therapeutic target.

With the advent of era of tumor immunotherapy,effect of intestinal flora in immunotherapy has attracted much attention.A large number of studies have confirmed the role of traditional Chinese medicine in regulating intestinal flora.Based on this,this paper discusses and summarizes the effect and mechanism of traditional Chinese medicine intervention on tumor immune checkpoint inhibitors(ICIs)treatment,providing a theoretical basis for in-depth understanding of the correlation between traditional Chinese medicine,intestinal flora and immunotherapy,and also providing new ideas for improving the efficacy of tumor immunotherapy and reducing immune-related adverse reactions.

Objective:To discuss the associations of mismatch repair(MMR)in gastric cancer with preoperative inflammatory indicators and clinicopathological features in the gastric cancer patients,and to construct a gastric cancer MMR predictive model based on preoperative inflammatory indicators and clinicopathological features of the gastric cancer patients,and to provide new ideas for evaluation of MMR status in gastric cancer.Methods:The data of 254 gastric cancer patients who underwent surgical treatment from September 2020 to October 2023 were included.According to the expression of MMR protein,the patients were divided into MMR normal(proficiout MMR,pMMR)group and MMR deficient(dMMR)group.The preoperative inflammatory indicators and clinicopathological features data of the gastric cancer patients in two groups were collected.The associations between inflammatory indicators,clinicopathological features,and MMR in dMMR group and pMMR group were analyzed using Chi-square test.The independent predictive factors for dMMR were selected to construct the nomogram.Receiver operating characteristic(ROC)curve and calibration curve were used to evaluate the predictive efficacy,and decision curve was used to evaluate the practicality of the predication model.Results:A total of 254 gastric cancer patients were included in the study,with 221 patients(87％)in pMMR group and 33 patients(13％)in dMMR group.There were statistically significant differences(P＜0.05)in age,tumor location,tumor differentiation degree,maximum tumor diameter,platelet-to-lymphocyte ratio(PLR),alkaline phosphatase(AKP),alkaline phosphatase-to-albumin ratio(AAR),fibrinogen(FB)-to-lymphocyte(FLR),FB-to-albumin(AL)(FAR),D-dimer(D-D),and FB of the gastric cancer patients between dMMR group and pMMR group.Univariate and multivariate Logistic regression analysis revealed maximum tumor diameter[odd ratio(OR)=2.958,95％confidence interval(CI):1.196-7.314,P=0.019],tumor location(OR=4.013,95％CI:1.596-10.089,P=0.003),tumor differentiation(OR=3.006,95％CI:1.250-7.230,P=0.014),FAR(OR=2.793,95％CI:1.179-6.616,P=0.020),and carbohydrate antigen 199(CA199)(OR=0.279,95％CI:0.084-0.929,P-0.038)were the independent predictors of dMMR.The area under the ROC curve(AUC)value of the gastric cancer MMR prediction model constructed based on inflammatory indicators and clinical pathological characteristics was 0.800 with the sensitivity of 0.851 and the specificity of 0.606.The calibration curve of the nomogram was found to fit the ideal curve well,and in Hosmer-Lemeshow test P=0.412,the clinical decision curve showed a better net benefit.Conclusion:The preoperative inflammatory indicators and clinicopathological features are associated with MMR in gastric cancer;maximum tumor diameter,tumor location,tumor differentiation,CA199,and FAR are the independent predictors of dMMR.The prediction model based on the above predictors could predict the MMR status of the dMMR gastric cancer patients.

Objective To explore the expression profile of adenosine A1 receptor in the paraventricular thalamic nucleus(PVT)in wakefulness/sleep state and its role in regulating sleep.Methods Male C57BL/6J mice(6～12 weeks old,weighing 22～28 g)were randomly divided into ZT0,ZT6,ZT10,ZT12,and sleep deprivation and recover0y sleep groups(n=16 or n=4).RT-qPCR and fluorescence in situ hybridization were used to observe the changes of adenosine A1 receptor in PVT.Whole-cell patch-clamp recording was conducted to determine the effects of adenosine on the membrane potential and discharge frequency of PVT neurons.The experimental animals were divided into adenosine A1 receptor interference group(n=8)and interference control group(n=7).After RNA interference,electroencephalography(EEG)and electromyography(EMG)were applied simultaneously to observe the changes in wake and sleep time between the 2 groups during sleep deprivation for 6 h and sleep recovery for 4 h.Results RT-qPCR and fluorescence in situ hybridization confirmed that the expression of adenosine A1 receptor in PVT was affected by wakefulness/sleep state,with the level in the ZT0 group(active stage)significantly higher than that in the ZT12 group(non-active stage,P<0.05).Whole-cell patch clamp recording indicated that adenosine exerted inhibitory effects on PVT neurons through 2 distinct response types(P<0.05),and the inhibition was in a diminishing trend with a decrease in the expression level of adenosine A1 receptor.After sleep deprivation,the expression level of adenosine A1 receptor showed significant intergroup differences:the level was significantly higher in the sleep deprivation group than the recovery sleep group(P<0.05),while that of the recovery sleep group was higher than that of the ZT6 group and that of the ZT10 group(P<0.05).After knocking down adenosine A1 receptor(shRNA-A1R)in the PVT,the wakefulness timing of the shRNA-A1R group was significantly increased,while the sleep timing was significantly decreased within 1 h after sleep recovery when compared with the interference control group(P<0.05).Conclusion The expression of adenosine A1 receptor in the PVT is dynamically regulated by sleep pressure,which was increasing as sleep pressure rises.

Objective:To investigate the incidence of collateral circulation in high-risk patients with sleep apnea and stroke treated by continuous positive airway pressure （CPAP） ventilation and to analyze the influencing factors. Methods:A total of 152 patients diagnosed with obstructive sleep apnea-hypopnea syndrome （OSAHS） combined with acute ischemic stroke （AIS） who were admitted to our hospital from January 2020 to June 2022 were selected for this study. Based on the apnea-hypopnea index （AHI）, the patients were divided into three groups: mild （n=44）, moderate （n=72）, and severe （n=36）. After treatment, the patients were further classified into a group without collateral circulation （n=30） and a group with collateral circulation （n=26）, which included those with moderate collateral circulation （n=69） and good collateral circulation （n=27）. Clinical data across the different groups were compared, and multiple factor analysis was performed to identify factors affecting the occurrence of collateral circulation. Results:The AHI and IL-6 levels in the severe group were significantly higher than those in the mild and moderate groups, while the levels of NO and PO2 were significantly lower in the severe group compared to the mild and moderate groups, with statistically significant differences among the three groups （P<0.05）. After treatment, all groups showed improvement, and the proportion of patients with collateral circulation was 84.09% in the mild group, 81.94% in the moderate group, and 72.22% in the severe group. Significant differences in age, AHI, NIHSS, NO, MoCA, and MMSE scores were observed between the groups with and without collateral circulation （P<0.05）. In the group with collateral circulation, the scores for age, AHI, and NIHSS in the good collateral circulation subgroup were significantly lower than those in the poor collateral circulation and moderate collateral circulation subgroups, while the scores for NO, MoCA, and MMSE were significantly higher in the good collateral circulation subgroup. Multi-factor analysis revealed that age, AHI, and NIHSS were independent risk factors for collateral circulation, whereas NO, MoCA, and MMSE served as protective factors that were negatively correlated with collateral circulation. Classification tree model results indicated that AHI had the greatest influence on the occurrence of collateral circulation among the five influencing factors, demonstrating good predictive capability. Conclusion:Most high-risk patients with sleep apnea and stroke are likely to develop collateral circulation following continuous positive airway pressure ventilation. Factors such as age, AHI, NIHSS, NO, MoCA, and MMSE are important determinants affecting the occurrence of collateral circulation.
Humans ; Collateral Circulation ; Continuous Positive Airway Pressure ; Stroke/physiopathology* ; Sleep Apnea, Obstructive/physiopathology* ; Risk Factors ; Male ; Incidence ; Female ; Middle Aged ; Aged ; Sleep Apnea Syndromes/physiopathology* ; Interleukin-6/blood*

Evidence-based treatment strategies for patients with cardiovascular disease and diabetes have been updated in recent years. However, substantial gaps remain between guideline recommendations and clinical practice, which justify the urgent need to improve the quality of care for patients with these conditions. The Chinese Society of Cardiology and the Chinese Society of Diabetes, in collaboration with the American Heart Association and the American Diabetes Association, designed the China Diabetes Cardiovascular project. The China Diabetes Cardiovascular project is a nationwide registry study aimed at improving the quality of care for patients with acute coronary syndrome and diabetes in China. Launched in 2021, this project has enrolled 36 hospitals across mainland China. Patients with a primary discharge diagnosis of comorbid acute coronary syndrome and diabetes will be eligible to participate. Pre-defined performance measures will be adopted to evaluate the quality of care for these patients. Multiple quality improvement strategies will be adopted, including providing monthly quality reports based on these measures, conducting a series of training courses, and distributing educational materials. A comprehensive dataset, encompassing patients' characteristics, medical history, treatment before and during the current hospitalization, and discharge medications for secondary prevention, will be collected through a web-based data collection platform. This project has the potential to improve the quality of care and reduce the care disparities in the management of patients with these diseases. Moreover, with its comprehensive data collection, this project will provide a strong foundation for exploring key clinical questions.

Objective To explore the anesthesia management experience in the interventional treatment of pediatric congenital heart diseases (CHD) percutaneously guided by transthoracic echocardiography (TTE) on a mobile operating platform. Methods From March to July 2023, a total of 13 patients from remote areas underwent interventional treatment for CHD on the mobile operating platform of Fuwai Yunnan Cardiovascular Hospital. Patients who received non-tracheal intubation general anesthesia were retrospectively included. Results Eight children who had difficulty cooperating with the surgery (due to young age, emotional tension, crying) received monitored anesthesia care with local anesthesia supplemented by sedative and analgesic drugs while maintaining spontaneous breathing under the monitoring and management of an anesthesiologist (i.e., non-tracheal intubation general anesthesia). Among them, there were 5 males and 3 females, with an age of (6.95±3.29) years and a body weight of (19.50±6.04) kg. Through transthoracic echocardiography, they were diagnosed with atrial septal defect (6 patients), residual shunt after patent ductus arteriosus ligation (1 patient), and severe pulmonary valve stenosis (1 patient). The surgery proceeded smoothly, with satisfactory anesthesia and surgical effects, complete analgesia, and satisfactory postoperative recovery. There was 1 patient of body movement and 1 patient of respiratory depression during the operation, and both patients completed the surgery successfully after treatment. All children had no serious surgery- and anesthesia-related complications. The anesthesia time was 40.5 (34.5, 47.5) min, the surgery time was 39.0 (33.0, 45.5) min, and the recovery time was 43.0 (28.0, 52.5) min Conclusion Interventional surgery for CHD guided by TTE at a mobile platform is a minimally invasive approach without radiation damage. Non-tracheal intubation general anesthesia with spontaneous breathing can be safely and effectively implemented in children who cannot cooperate.

Objective To establish a method for acquisition, perfusion, preservation and transportation of the genetically modified pig kidneys. Methods An eight genetically modified pig was utilized as experimental subject. Prior to kidneys procurement, the health status of the pig was assessed through hematology examination, and the vascular structure of the kidneys was examined using imaging techniques. Following kidneys acquisition, the pig kidneys were perfused and subsequently packaged into the cryogenic storage container labeled "For Organ Transportation Only" for interprovincial transport after communicating the transportation process with transportation department. To evaluate pathological damage to the pig kidneys, a serious of methods were employed such as hematoxylin-eosin (HE) staining, real-time fluorescent quantitative polymerase chain reaction (RT-qPCR), terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) fluorescence staining and enzyme-linked immune absorbent assay (ELISA). Results The preoperative examination of the eight genetically modified pig showed that the serum creatinine was 73.2 μmol/L, blood urea nitrogen was 2.8 mmol/L and hemoglobin was 116 g/L, all within the normal range, indicating normal renal function. CT angiography revealed no lesions in the pig kidneys, and no dilation, stenosis or premature branching of the blood vessels. The total time of obtaining the left and right kidneys from the eight genetically modified pig was (125 ± 10) min, with a blood loss of (20 ± 2) mL. The warm ischemia times were 3 min and 7 min, respectively. The perfusion and trimming times of the left and right kidneys were 36 min and 41 min, respectively. After perfusion, both kidneys were white and moist. The cold preservation and transportation time was 8 h. HE staining showed that some glomeruli were shrunk, and the lumens of the surrounding renal tubules were slightly depressed and swollen with partial inner membrane shedding and microvacuoles formed when the kidneys were preserved for 8 h. The level of cysteinyl aspartate-specific proteinase-3 messenger RNA in the kidneys tissue gradually increased with the extension of cold preservation time after 2 h (P<0.05). TUNEL fluorescence staining showed that only a small number of cells underwent apoptosis after 8 h of cold preservation, which was not significantly different from that at 0 h (P>0.05). ELISA results showed that the contents of lactate dehydrogenase (LDH) and creatinine in the preservation solution remained relatively stable, but the content of kidney injury molecule 1 (KIM-1) gradually increased with the extension of preservation time, suggesting that the pig kidneys had mild injury. Conclusions By establishing methods for acquisition, perfusion, preservation and transportation of the kidneys from genetically modified donor pig, it is possible to effectively and reliably use genetically modified pig kidneys for xenotransplantation.

[Objective] To establish a method for detecting the potency of recombinant human coagulation factor Ⅶa for injection. [Methods] By adding the sample and factor Ⅶ deficient plasma to the sample cup and activating the reaction with prothrombin time assay reagent (PT reagent), the coagulation time of the sample was determined by the change in magnetic bead swing amplitude in the sample cup. The logarithm of coagulation time was inversely proportional to the logarithm of human factor Ⅶa potency. [Results] Under the experimental conditions, the specificity of the methodology was evaluated through spiked recovery, and the recovery rates ranged from 90.0% to 110.0%. Within the range from 0.125 to 1.000 IU/mL, there was a good linear response between the potency and coagulation time of the standard and sample, with correlation coefficients r>0.99. As for the accuracy and repeatability, the recovery rates of various concentrations detected in the stock solution were 101.0%, 100.0% and 112.0%, respectively, with RSD values of 2.6%, 4.0% and 0.0%, respectively. The recovery rates of various concentrations in finished product testing were 104.0%, 94.7% and 112.0%, respectively, with RSD values of 1.9%, 2.4% and 0.0%, respectively. As for the intermediate precision, the RSD were 4.5% and 3.7%, respectively. After treated with sample diluent, the sample was tested at room temperature for 6 hours and still exhibited relatively stable biological activity. [Conclusion] This detection method is accurate, stable, easy to operate and highly automated, and is suitable for detecting the potency of recombinant human coagulation factor Ⅶa for Injection.