Objective To investigate the role of β-arrestin2 in intestinal inflammation and illustrate the mechanisms from the perspective of epithelial barrier function. Methods Dextran sodium sulfate(DSS)is used to induce acute intestinal colitis in mice. The experiment groups are designed as the wild type control(WT),the wild type colitis (WT+DSS) and the β-arrestin2- knockout colitis (KO+DSS). The expression of β-arrestin2 gene by mRNA and protein level is compared between the WT and WT + DSS groups. The difference of weight loss , disease activity index(DAI),spleen weight,colon length,histological score,intestinal permeability and important tight junction proteins (occludin ,claudin1 and ZO-1) were detected in the WT+DSS and KO+DSS groups. Results Compared with the WT group,the expression of β-arrestin2 was significantly higher in the colon of the WT+DSS group. Compared with the WT+DSS group,the KO+DSS group had less weight loss(P < 0.05),lower DAI(P<0.05),smaller spleen,longer colon and lower histological score(P=0.002). The KO+DSS group had a lower intestinal permeability(P = 0.009)and higher protein level of occludin and claudin1.There was no signifi-cant difference of ZO-1 in the two groups. Conclusion β-arrestin2 may promote mouse colitis through impairment of epithelial barrier function.

Alemtuzumab (Campath) was successfully injected in 21 kidney transplant patients,7 islet transplant patients and 1 simultaneous kidney and islet transplant patient for either prevention or treatment of graft rejection.Prophylactic administration was successfully completed in all patients without discontinuation.Adverse events were not observed in 11 patients (38%),but hypertension in 18 patients (62%),shivering in 3 patients (10.3%),high fever in 3 patients (10.3%),and bronchospasm in 1 patient (3%),respectively.All complications alleviated after proper therapy.During the prophylactic administration of alemtuzumab,strict,timely and proper ward-management was needed.Care for lung,perineum,skin,diet and psychological nursing were necessary.Neither graft acute rejection nor graft chronic rejection episode occurred in all patients during 6 months to 2 years follow-up.Therefore,long term effects of Alamtuzumab and consequences of lymphocytopenia need further observation.

Objective To investigate the expressions of Nanog and Oct4 (stem cell transcription factors) in endometriosis and adenomyosis, and to explore their potential functions in the development of endometriosis and adenomyosis. Methods The expressions of Nanog and Oct4 in the ectopic and eutopic endometrium of 50 patients with endometriosis and/or adenomyosis (ectopic endometrium group and eutopic endometrium group), and 21 patients free from endometriosis and adenomyosis (control group) were detected by immunohistochemical SABC methods. Statistical analysis was conducted for the correlation between the expressions of Nanog and Oct4 based on patients′ clinical pathological parameters. Results Nanog and Oct4 protein expressions in ectopic endometrium group were higher than that in control group (P<0.01);the expressions of Nanog and Oct4 in eutopic endometrium group and control group showed no significance (P > 0.05); there was positive correlation between the expressions of Nanog and Oct4 in ectopic endometrium group (r = 0.590, P < 0.01). Conclusion Nanog and Oct4 present high expression in eutopic and ectopic endometrium , which may play a important role in the development of endometriosis and adenomyosis.

Objective:To identify the risk factors for acute kidney injury (AKI) after non-cardiac surgery.Methods:Medical records of patients who underwent non-cardiac surgery with general anesthesia in our hospital from October 1, 2016 to March 31, 2021 were collected.AKI was diagnosed using the Kidney Disease Improving Global Outcomes definition of AKI.Multi-factor logistic regression analysis was used to screen the risk factors affecting AKI.Generalized linear regression was used to analyze the factors influencing the difference in serum creatinine before and after surgery.Results:A total of 2 214 patients were eventually enrolled, and the incidence of AKI was 5.15%.The results of multi-factor logistic regression analysis showed that preoperative hypertension, American Society of Anesthesiologists Physical Status classification ≥ Ⅲ, intraoperative bleeding >300 ml, emergency surgery, and grade IV surgery were independent risk factors for AKI after non-cardiac surgery ( P<0.05). The results of generalized line regression analysis showed that preoperative hyponatremia, intraoperative bleeding >300 ml, emergency surgery, and duration of anesthesia were positively correlated with increased difference in serum creatinine before and after surgery ( P<0.05). Conclusions:Preoperative hypertension, American Society of Anesthesiologists Physical Status classification ≥ Ⅲ, intraoperative bleeding >300 ml, emergency surgery and grade IV surgery are independent risk factors for AKI after non-cardiac surgery.

Objective:To explore the key genes and its biological functions of aldosterone producing adenoma (APA) using bioinformatics analysis.Methods:Differentially expressed genes of APA were identified from two training datasets GSE60042 and GSE64957 in GEO database. Function and pathway enrichment analyses for differentially expressed genes were performed and transcriptional regulation network among these genes were determined. Hub genes were extracted by node analysis from the protein-protein interaction (PPI) network. The expression of key genes was verified by a testing dataset GSE8514. Receiver operating characteristic(ROC) curve analysis was applied to assess the diagnostic efficiency of key genes in APA. The biofunction of each key gene were determined by gene set enrichment analysis (GSEA).Results:A total of 68 differentially expressed genes, including 33 up-regulated genes and 35 down-regulated genes, were detected from the training datasets. These genes were mainly enriched in aldosterone biosynthetic process, calcium signaling pathway, serotonin receptor signaling pathway, transcriptional activator activity, and regulation of transcription. JUN and VDR were at the center of the transcriptional factor-gene network. Furthermore, we identified nine Hub genes from the PPI network. In testing dataset, CYP11B2 and VDR showed the higher expression, while JUN, NFKBIZ, EGR3, and KLF6 showed lower expression in APA (all P<0.05), and the value of area under ROC curve analysis was 0.936, 0.833, 0.953, 0.854, 0.868, and 0.929, respectively. GSEA indicated the alter of key genes in APA led to up-regulation of the steroid biosynthesis, cell adhesion molecules, immune cells signaling pathway, and complement and coagulation cascades [all normalized enrichment score (NES)>1.5, P<0.05], but down-regulation of the DNA replication, ribosome, and autophagy (all NES<-1.5, P<0.05). Conclusion:Results of bioinformatics indicate that JUN and VDR are key transcriptional factors, and CYP11B2, NFKBIZ, EGR3, and KLF6 are the key genes for APA, which are involved in the steroid biosynthesis, cell adhesion molecules, immune cells signaling pathway in APA.

The antigen gene expression level of a DNA vaccine is the key factor influencing the efficacy of the DNA vaccine. Accordingly, one of the ways to improve the antigen gene expression level of a DNA vaccine is to utilize a plasmid vector that is replicable in eukaryotic cells. A replicative DNA vaccine vector pCMVori was constructed based on the non-replicative pcDNA3.1 and the replicon of porcine circovirus 2 (PCV2) in this study. An EGFP gene was cloned into pCMVori and the control plasmid pcDNA3.1. The two recombinant vectors were transfected into PK-15 cell, and the plasmid DNA and RNA were extracted from the transfected cells. Real-time PCR was used to determine the plasmid replication efficiency of the two plasmids using plasmid before and after Bcl Ⅰ digestion as templates, and the transcription level of the Rep gene in PCV2 replicon was detected by RT-PCR. The average fluorescence intensity of cells transfected with the two plasmids was analyzed with software Image J, and the transcription level of EGFP was determined by means of real-time RT-PCR. The results showed that the replication efficiency of pCMVori in PK-15 cells incubated for 48 h was 136%, and the transcriptions of Rep and Rep' were verified by RT-PCR. The average fluorescence intensity of the cells transfected with pCMVori-EGFP was 39.14% higher than that of pcDNA3.1-EGFP, and the transcription level of EGFP in the former was also 40% higher than that in the latter. In conclusion, the DNA vaccine vector pCMVori constructed in this study can independently replicate in eukaryotic cells. As a result, the expression level of cloned target gene was elevated, providing a basis for developing the pCMVori-based DNA vaccine.
Animals ; Swine ; Circovirus/genetics* ; Vaccines, DNA/genetics* ; Replicon/genetics* ; Genetic Vectors/genetics* ; Plasmids/genetics*

Objective:To explore the survival prognosis of hepatocellular carcinoma patients over 60 years old with comorbidities treated with microwave ablation.Methods:A retrospective analysis of 267 patients with hepatocellular carcinoma aged 60 years or older admitted to the PLA General Hospital from April 2012 to September 2022 were analyzed, including 179 patients with preoperative comorbidities and 88 patients without comorbidities. The overall survival (OS) and progression-free survival (PFS) of the two groups were compared by the Log-rank test, and the Cox proportional hazards regression model was used to evaluate ablation-related risk factors.Results:A total of 267 patients were included (comorbidity group, n=179; no comorbidity group, n=88). There were no statistical differences in OS and PFS between the two groups (all P>0.05). Multivariate Cox regression analysis showed that comorbidities were not risk factors that affected the survival prognosis (OS and PFS) of patients with hepatocellular carcinoma after microwave ablation ( P>0.05). Total bilirubin (hazard ratio 0.356, 95% CI=0.174-0.731, P=0.005) was a risk factor affecting OS; tumor number (hazard ratio 0.538, 95% CI=0.365-0.793, P=0.002) and international coagulation normalized ratio (hazard ratio 1.022, 95% CI=1.001-1.043, P=0.040) were risk factors affecting PFS. Subgroup analysis showed that patients with a maximum diameter of >3 cm and female patients, the OS of the comorbidity group was significantly lower than that of the non-comorbidity group( P<0.05). Conclusions:Microwave ablation therapy remains an effective treatment modality in hepatocellular carcinoma patients over 60 years of age with comorbidities, and its survival prognosis is not inferior to patients with hepatocellular carcinoma without comorbidities.

ObjectiveTo investigate the effects of Aconiti Coreani Radix and Typhonii Rhizoma on the urinary metabolites of gerbils with stroke by non-targeted metabolomics technique， and then to clarify the mechanism of the two， as well as their similarities and differences. MethodTwenty-four gerbils were randomly divided into control group（CG）， model group（MG）， Aconiti Coreani Radix group（RA） and Typhonii Rhizoma group（RT）. Except for the CG， ischemic stroke model was constructed using right unilateral ligation of gerbil carotid artery in the remaining groups. Except for the CG and MG， rats in the other groups received whole powder suspension（0.586 mg·g^-1） was administered for 14 days. The neurological deficit in each group was scored by Longa scoring on days 0， 3， 7 and 14. After the end of administration， the serum， brain tissue and urine of gerbils in each group were collected， and the rate of cerebral infarction was detected by 2，3，5-triphenyltetrazolium chloride（TTC）， and the levels of interleukin（IL）-6， tumor necrosis factor（TNF）-α， malondialdehyde（MDA）， superoxide dismutase（SOD）， glutathione（GSH）， and nitric oxide（NO） in serum and brain tissue were determined by enzyme-linked immunosorbent assay（ELISA）. The urine metabolomics of gerbils in each group was studied by ultra performance liquid chromatography-quadrupole-electrostatic field orbitrap high resolution mass spectrometry（UPLC-Q-Orbitrap-MS）， and the data were processed by multivariate statistical analysis， and differential metabolites were screened based on value of variable importance in the projection（VIP） of the first principal component>1 and t-test P<0.05. Metabolic pathway analysis of the screened differential metabolites was performed using Kyoto Encyclopedia of Genes and Genomes（KEGG） database and Metaboanalyst 5.0. ResultCompared with the CG， the neurological deficit score was significantly increased in the MG（P<0.05）， compared with the MG， the neurological deficit scores in the RA and RT were significantly reduced after 7 d and 14 d（P<0.05）. Compared with the CG， the rate of cerebral infarction was significantly increased in the MG（P<0.05）， compared with the MG， the rates of cerebral infarction in the RA and RT were significantly reduced（P<0.05）. Compared with the CG， the levels of IL-6， TNF-α， and MDA in the serum and brain tissue of gerbils from the MG were significantly increased（P<0.05）， and the levels of SOD， GSH and NO were significantly reduced（P<0.05）. Compared with the MG， Aconiti Coreani Radix and Typhonii Rhizoma could down-regulate the levels of IL-6， TNF-α and MDA， and up-regulated the levels of SOD， GSH and NO. A total of 112 endogenous differential metabolites were screened by urine metabolomics， of which 16 and 26 metabolites were called back by Aconiti Coreani Radix and Typhonii Rhizoma， and could be used as potential biomarkers for both treatments in stroke gerbils， respectively. The results of the pathway analysis showed that both Aconiti Coreani Radix and Typhonii Rhizoma had regulatory effects on arginine and proline metabolism， pyrimidine metabolism， and aminoacyl-tRNA biosynthesis. In addition， Aconiti Coreani Radix could also regulate riboflavin metabolism， Typhonii Rhizoma could also regulate purine metabolism， glycine， serine and threonine metabolism， arachidonic acid metabolism， biosynthesis of pantothenate and coenzyme A， and β-alanine metabolism. ConclusionBoth Aconiti Coreani Radix and Typhonii Rhizoma have better therapeutic effects on stroke， with Aconiti Coreani Radix having stronger effects. From the metabolomics results， the main metabolic pathways regulated by Aconiti Coreani Radix involve amino acid metabolism， oxidative stress and so on， while Typhonii Rhizoma mainly involve amino acid metabolism， lipid metabolism， energy metabolism， etc.