ObjectiveTo explore the application value of probe-based confocal laser endomicroscopy (pCLE) in rapidly detecting and evaluating the morphological characteristics of digestive tract tissues in mice. MethodsTwelve male SPF Kunming mice aged 6 weeks were randomly divided into two groups. Six mice were subjected to gastric gavage with 52% Red Star Erguotou to establish the model, and six were given saline by gastric gavage as a control. After 28 days of modeling, 3 mice were randomly selected from each group. After deep anesthesia induced by inhalation of 3% isoflurane, the mice were sacrificed by cervical dislocation. The stomach, duodenum, jejunum, and rectum tissues were excised and immersed in 1% fluorescein sodium solution for staining. The microstructure of the mucosal surface of each tissue was observed using pCLE. The remaining mice in the model group and the control group were deeply anesthetized by inhaling 3% isoflurane, then cardiac perfusion was performed successively with saline and 4% paraformaldehyde. The stomach, duodenum, jejunum, and rectum tissues were excised for dehydration, section and hematoxylin-eosin (HE) staining, and the morphological changes of the tissues were observed under a microscope. ResultsUnder pCLE imaging, fluorescence staining on the surface of the gastrointestinal mucosa was uniform in the control group; the morphology of gastric pits, intestinal villi, and intestinal crypts was intact, arranged compactly, and had distinct boundaries. In the model group, the gastrointestinal mucosa exhibited mucosal swelling and deformation, with uneven fluorescence staining and fluorescein leakage. Furthermore, some tissues showed defects or cell shedding, and the boundaries between adjacent characteristic structures (e.g., gastric pits, intestinal crypts) were blurred. HE staining showed that the gastrointestinal tissue structure of the control group mice was normal and well-organized, with no structural defects. Moreover, submucosal glands were uniform in size, with no hyperplasia observed, and no obvious inflammatory cell infiltration. In the model group, some gastrointestinal mucosal structures were defective and sparsely arranged; submucosal glands showed atrophy, accompanied by obvious inflammatory cell infiltration. The histological characteristics detected by pCLE were consistent with those of HE staining. ConclusionpCLE can be used to obtain rapid, real-time, large-scale, and high-resolution microscopic imaging of the gastrointestinal mucosa, realistically and comprehensively displaying its physiological and microstructural characteristics. It shows promising prospects and practical utility in the histological evaluation of digestive system injuries in small animals.

The innate immune sensor NLRP3 inflammasome overactivation is involved in the pathogenesis of ulcerative colitis. PGAM5 is a mitochondrial phosphatase involved in NLRP3 inflammasome activation in macrophages. However, the role of PGAM5 in ulcerative colitis and the mechanisms underlying PGAM5 regulating NLRP3 activity remain unknown. Here, we show that PGAM5 deficiency ameliorates dextran sodium sulfate (DSS)-induced colitis in mice via suppressing NLRP3 inflammasome activation. By combining APEX2-based proximity labeling focused on PGAM5 with quantitative proteomics, we identify NEK7 as the new binding partner of PGAM5 to promote NLRP3 inflammasome assembly and activation in a PGAM5 phosphatase activity-independent manner upon inflammasome induction. Interfering with PGAM5-NEK7 interaction by punicalagin inhibits the activation of the NLRP3 inflammasome in macrophages and ameliorates DSS-induced colitis in mice. Altogether, our data demonstrate the PGAM5-NEK7 interaction in macrophages for NLRP3 inflammasome activation and further provide a promising therapeutic strategy for ulcerative colitis by blocking the PGAM5-NEK7 interaction.

Objective To observe the value of motion freeze technology(MFT)for improving the quality of cone-beam CT(CBCT)images of renal artery aneurysms.Methods Twenty patients with renal artery aneurysms who would receive transcatheter arterial embolization were prospectively enrolled.Renal artery angiography was performed under the guidance of CBCT,and motion artifacts were processed using MFT.Then subjective and objective evaluation of images before and after MFT correction were performed,aimed on displaying of renal artery,parent artery and vascular tree in target area,and the mean value(mean)and standard deviation(SD)of unit density pixels,margin sharpening error rate of blood vessels were measured and calculated.Results After MFT correction,the displaying of renal artery on maximum intensity projection images and parent artery on reconstructed CBCT images were significantly better than those before correction(both P＜0.05),clear vascular trees could be automatically extracted using Flightplan for Liver software,SD and margin sharpening error rate of blood vessels of renal artery aneurysm significantly decreased but mean significantly increased on CBCT images(all P＜0.05).Conclusion MFT could effectively improve the quality of CBCT images of renal artery aneurysms.

RNA modifications affect many biological processes and physiological diseases. The 5-methylcytosine (m⁵C) modification regulates the progression of multiple tumors. However, its characteristics and functions in hepatocellular carcinoma (HCC) remain largely unknown. Here, we found that HCC tissues had a higher m⁵C methylation level than the adjacent normal tissues. Transcriptome analysis revealed that the hypermethylated genes mainly participated in the phosphokinase signaling pathways, such as the Ras and PI3K-Akt pathways. The m⁵C methyltransferase NSUN2 was highly expressed in HCC tissues. Interestingly, the expression of many genes was positively correlated with the expression of NSUN2, including GRB2, RNF115, AATF, ADAM15, RTN3, and HDGF. Real-time PCR assays further revealed that the expression of the mRNAs of GRB2, RNF115, and AATF decreased significantly with the down-regulation of NSUN2 expression in HCC cells. Furthermore, NSUN2 could regulate the cellular sensitivity of HCC cells to sorafenib via modulating the Ras signaling pathway. Moreover, knocking down NSUN2 caused cell cycle arrest. Taken together, our study demonstrates the vital role of NSUN2 in the progression of HCC.
Humans ; ADAM Proteins ; Apoptosis Regulatory Proteins ; Carcinoma, Hepatocellular/metabolism* ; Liver Neoplasms/metabolism* ; Membrane Proteins ; Methyltransferases/metabolism* ; Phosphatidylinositol 3-Kinases/metabolism* ; Repressor Proteins ; RNA, Messenger/metabolism* ; Sorafenib ; RNA Methylation/genetics*

Objective:To evaluate the utility of CT perfusion (CTP) for assessment of effect of encephalo-duro-arterio-synangiosis (EDAS) on moyamoya disease (MMD).Methods:Thirty-eight adult MMD patients, who underwent EDAS in our hospital from March 2014 to October 2019, were chosen in our study. All patients received CTP and digital subtraction angiography (DSA) before and after surgery; cerebral blood flow (CBF), cerebral blood volume (CBV), mean transit time (MTT) and time to peak (TTP) in the operative cerebral cortices and their contralateral mirror areas, as well as brainstems were measured respectively; their relative parameter values (rCBF, rCBV, rMTT and rTTP) were calculated with the brainstem as the reference; perfusion changes of cerebral tissues were observed before and after surgery, and efficacies of these parameters in evaluating the surgical treatment effect were compared. According to the degrees of collateral formation detected by postoperative DSA, the 42 hemispheres of these patients were divided into group of good collateral formation and group of poor collateral formation, and the differences of perfusion changes in the two groups were further discussed.Results:CTP revealed that cerebral perfusion at surgical side after EDAS was significantly improved in all the 42 hemispheres; significantly increased rCBF, and significantly decreased rMTT and rTTP were noted as compared with those before surgery ( P<0.05). Receiver operating characteristic (ROC) curve showed that the area under the curve of rTTP (0.897) was the largest among all parameters, with diagnostic sensitivity of 87.5% and specificity of 80.8%. Twenty-six patients (61.9%) were into the group of good collateral formation and 16 (38.1%) were into the group of poor collateral formation; changed values of rCBF and rMTT in the group of good collateral formation were significantly better than those in the group of poor collateral formation ( P<0.05). Conclusion:CTP can quantitatively evaluate cerebral hemodynamic changes after EDAS in moyamoya disease patients, and postoperative rTTP changes can reflect the degrees of collateral vessel formation.

Hypothyroidism is a common adverse reaction in clinical multi-target tyrosine kinase inhibitor (TKI) and may be used as a predictor of efficacy of TKI drugs.Studies show that hypothyroidism is significantly associated with good prognosis in patients with renal cell carcinoma treated with multi-target TKI,which may be related to inhibition of vascular endothelial growth factor receptor pathway,potential off-target effects and inhibition of cell proliferation.Whereas in the patients with hepatocellular carcinoma who received multi-target TKI therapy,hypothyroidism is an indicator of poor prognosis,which is related to the T3/TR pathway regulating liver cancer growth and invasion.The predictive function and mechanism of thyroid hormone levels in different tumor patients receiving multi-target TKI drugs are different,and further research is still needed.

Objective:To investigate whether facial expression recognition correct rates were changed in stable patients with major depressive disorder.Methods:A cross-sectional study was performed in patients with major depressive disorder (n=80) (according to the DSM-IV) and matched normal controls by age, gender and education year (n=41) in this study.All subjects would recognize five different expression (anger, contempt, fear, happiness and sadness).These emotions were displayed on four different male and female computerized cartoon faces, and each expression was shown once in a static format and once in a dynamic format.The facial expression recognition correct percentage would be compared between the two groups.Results:The correct identification rate of contempt was lower in the patients than in the normal controls (93％ vs.94％, P＜0.05).The correct identification rate of dynamic contempt was lower in the patients than in the normal controls (93％ vs.95％, P＜0.05).The correct identification rate of fear was lower in the patients than in the normal controls (89％ vs.93％, P＜0.05).The correct identification rate of dynamic fear was lower in the patients than in the normal controls (90％ vs.94％, P＜0.05).Regarding error rates of misattribution, compared with normal controls, depressive patients over-identified fear to anger (3.7％vs.1.8％) and contempt (2.9％vs.0.9％), sadness to contempt (5.0％ vs.1.7％) and happiness (2.0％ vs.0.2％) (Ps＜0.05).Conclusion:Patients with MDD have significantly great difficulties identifying negative expressions.

Objective:To investigate the natural course and psychosocial risk factors of depressive symptoms in college freshmen.Methods:Changes in depressive symptoms among college freshmen were observed for two year periods,self-reporting questionnaires were used to collect data every half yearly.The depressive symptoms were assessed with the Centre for Epidemiological Study-Depression Scale,and psychosocial factors were tested as the potential risk factors,rating by Zung Self-Rating Anxiety Scale,Eysenck Personality Questionnaire-Neuroticism Subscale,Rosenberg Self-Esteem Scale,Frost Multidimensional Perfectionism Scale,Parental Rearing Scale,Adolescents Self-Rating Life Events Checklist,Epworth Sleepiness Scale,daytime sleep time,Godin Leisure-Time Exercise Questionnaire.Generalized estimating equations model was used to analyze the date.Results:Totally 1339 college students were recruited in the survey.It was found the depressive scores significantly increased in the second and third surveys,then decreased in the fourth and fifth surveys.Male (Coef=-1.01,SE =0.42,P =0.017),not interested in the major (Coef=3.89,SE =1.42,P =0.006),neuroticism(Coef =0.79,SE =0.23,P =0.001),self-esteem(Coef =-1.57,SE =0.25,P ＜ 0.001),anxiety(Coef =4.79,SE =0.16,P ＜ 0.001),life events (Coef =0.08,SE =0.01,P ＜ 0.00l),daytime sleepiness (Coef =0.80,SE =4.56,P ＜ 0.001) were significantly correlated with depressive symptoms.Conclusion:The first year of college is a critical time in the prevention depression.Not interested in the major,high neuroticism,low self-esteem,anxiety,life events,daytime sleepiness may be risk factors of depressive symptom in college students.

Objective To evaluate the application value of glycerol blood chocolate soup in the bacteria preservation of Haemophilus influenzae.Methods Haemophilus influenzae strains ATCC10211,ATCC49247 and ATCC49766 were saved into glycerol blood chocolate soup,then it was stored in refrigerator at below -35℃.The preserved strains from the refrigerator at 1 month,3 months,6 months,1 year,2 years were taken out,and dissolved it at 35℃ for 5min,and then immediately transferred it to chocolate agar plate,placed it in the 35℃,CO2 environment overnight.Observed the survival of strains,colony morphology and whether the demand factor test had change.Saved the standard strains ATCC49247 and ATCC49766 2 years for the paper dispersion method drug sensitive test.Results After 2 years of observation,3 strains of Haemophilus influenzae strains grew well in the survival rate of 100%,the colony was colorless,transparent and flat moist,it could be observed gram small negative bacilli by Gram staining, blood agar satellite test was positive,no hemolysis,Columbia satellite test was negative.All traits were the characteristics of Haemophilus influenzae.The drug sensitivity characteristics of ATCC49247,ATCC49766 standard strains were still in line with the requirements of CLSI.Conclusion Glycerol blood chocolate soup is suit for Haemophilus influenzae bacteria preservation.

Objective To compare the clinical value of two kinds of imunoassay method including CLIA and ELISA in the detection of related antibody of patients with HCV.Methods 60 patients with HCV were chosen as study subjects,and the related antibodies of patients were detected by CLIA and ELISA.The positive rates of anti HCV antibody,anti-3E antibody,anti-GP210 antibody,anti-SP100 antibody,anti-PML antibody and anti-AMA -M2 antibody detected by CLIA and ELISA were compared.Results The positive rates of anti-HCV antibody detected by CLIA and ELISA were 83.33％ (50/60),98.33％ (59/60),respectively.The anti-HCV antibody detected by CLIA was significantly higher than by ELISA (x2 =4.73,P =0.03).The positive rates of anti-3E antibody,anti-GP210 antibody,anti-SP100 antibody,anti-PML antibody and anti-AMA-M2 antibody detected by ELISA were 11.67 ％ (7/60),6.67 ％ (4/60),11.67 ％ (7/60),10.00 ％ (6/60),11.67 ％ (7/60),respectively,those detected by CLIA were 73.33 ％ (44/60),50.00％ (30/60),38.33 ％ (23/60),58.33 ％ (35/60),80.00％(48/60),respectively.The positive rates of anti-3E antibody,anti-GP210 antibody,anti-SP1O0 antibody,anti PML antibody and anti-AMA-M2 antibody detected by CLIA were significantly higher than by ELISA(x2 =18.81,11.51,13.23,15.08,13.75,all P＜0.05).Concelusion Compared with ELISA,CLIA in the detection of related antibodies of patients with HCV can efficiently higher the positive rate of anti-HCV and autoantibody.