[ ABSTRACT] AIM: To investigate the differential expression of human leukocyte antigen-G ( HLA-G) isoforms and its receptors in human monocyte line THP-1 after human cytomegalovirus ( HCMV) infection for exploring the role of HLA-G in HCMV escaping the immune response of the organism .METHODS: THP-1 cells were infected with HCMV Towne strain.The expression of HLA-G isoforms at mRNA and protein levels was determined by RT-PCR and Western blot, respectively.The surface expression of HLA-G and its receptors ILT2/ILT4 and the cell viability were analyzed by flow cytometry.The levels of soluble HLA-G (sHLA-G) and IL-10 were measured by ELISA.RESULTS:After infection of the THP-1 cells with HCMV , no obvious apoptosis in the cells was observed , and the viability of the cells was high .A significant up-regulation of HLA-G1,-G3,-G4 and-G5 at mRNA expression level 1 d after infection was found , while the protein expression of HLA-G1 and HLA-G5 isoforms was mainly detected .The expression of HLA-G/ILT2/ILT4 was evi-dently up-regulated 1 d after infection .The level of sHLA-G was significantly increased 1 d after infection as compared with control group (P<0.01).The expression of IL-10 was obviously up-regulated 1 d post-infection as compared with control group.CONCLUSION:The differential expression of HLA-G isoforms and secretion of the receptors ILT 2/ILT4 and IL-10 in the THP-1 cells are induced after HCMV infection .This study provides experimental evidence for evaluating the immune mechanism of HCMV infection .

Objective To investigate the effect of glucagon-like peptide-1 (GLP-1) receptor agonist liraglutide on hypoxia/reoxygenation (H/R)-induced cardiomyocytes death under high glucose condition and the potential mechanisms.Methods H9C2 cardiomyocytes were divided into 4 groups:normal glucose (N,5 mmol/L),high glucose (G,20 mmol/L),high glucose in combination with liraglutide (L,100 nmol/L),high glucose in combination with liraglutide and wortmannin (W,25 nmol/L).The apoptosis of H9C2 was detected by TUNEL assay.Nitric oxide synthetase(eNOS),nitric oxide (NO) and reactive oxygen(ROS) in supernatants were measured by enzymatic analysis,p-PI3K,PI3K,p-Akt,Akt,Bcl-2,caspase-3 were examined by western blotting.Results Compared with cells in N group,the apoptosis of H9C2 cells induced by H/R was markedly increased [(15.79 ± 3.92) ％ vs (9.74 ± 1.14) ％,P =0.028] in G group.The same was true for ROS [(489.63 ±21.01) U/ml vs (338.50 ±43.60) U/ml,P ＜0.001] and caspase-3 levels (1.87 ±0.03 vs 1.15 ±0.04,P ＜0.001),but not for Bcl-2 protein expression (1.79 ± 0.06 vs 1.89 ±0.03,P =0.047).Pretreatment of cells with liraglutide (100 nmol/L) prevented the cell death induced by high glucose and H/R together with decrease of ROS and caspase-3 levels and increase of Bcl-1 expression.Moreover,treatment of cells with liraglutide also significantly increased phosphorylation of PI3K and Akt (p-PI3K/PI3K:0.87 ± 0.07 vs 0.59 ± 0.09,P =0.002;p-Akt/Akt:0.34 ± 0.01 vs 0.08 ± 0.01,P＜0.001),eNOS[(41.29 ±0.56) μmpl/L vs (37.20 ±0.52)μxmpl/L,P ＜0.001]and NO [(31.24 ±0.40) μmpl/L vs (26.66 ±0.53) μmpl/L,P ＜0.001] levels.Furthermore,addition of PI3K/Akt inhibitor wortmanin markedly inhibited the expression of p-PI3K/PI3K,p-Akt/Akt,reversed the changes of eNOS,NO,caspase-3 and Bcl-2 by liraglutide,and abolished the protective effect of liraglutide on cell apoptosis.Conclusions GLP-1 receptor agonist liraglutide treatment could alleviate cardiomyoeytes apoptosis induced by high glucose and H/R through the activation of PI3K-Akt-eNOS-NO signaling pathway and inhibition of oxidative stress.

Objective To investigate the predictive value of serum ferritin(SF)and glycated haemoglobin(HbA1c)in the occurrence of adverse perinatal outcomes in pregnant women with gestational diabetes melli-tus(GDM).Methods A total of 97 pregnant women with GDM who underwent antenatal examination and gave birth in Qionglai Maternal and Child Health Hospital from March 2021 to February 2023 were selected as observation group,and another 116 healthy pregnant women who came to the hospital for prenatal examina-tion were concurrently selected as control group.The fasting blood glucose(FBG),SF and HbA1c were com-pared between two groups,and the clinical characteristics,SF and HbA1c levels were compared among women with different perinatal outcomes.Then the influencing factors of adverse perinatal outcomes in GDM preg-nant women were identified,and the value of serum SF and HbA1c in predicting adverse perinatal outcomes in GDM pregnant women was determined.Results FBG,triglycerides(TG),total cholesterol(TC),low-density lipoprotein cholesterol(LDL-C),SF,and HbA1c were elevated in observation group compared to control group,and the differences were statistically significant(P＜0.05).FBG,TG,TC,SF,and HbA1c were higher in poor perinatal outcome group than in good perinatal outcome group,and the differences were statistically significant(P＜0.05).Multivariate Logistic regression analysis denoted that FBG,TG,TC,SF and HbA1c were independent influencing factors for adverse perinatal outcomes(P＜0.05).Receiver operating character-istic(ROC)curve analysis indicated that the area under the curve of single detection and combined detection of SF and HbA1c in predicting the adverse perinatal outcomes in GDM were 0.741(95％CI:0.638-0.844),0.685(95％CI:0.570-0.800)and 0.874(95％CI:0.797-0.951),respectively.Conclusion SF and HbA1c are abnormally elevated in pregnant women with GDM,and SF combined with HbA1c has certain predictive value for perinatal outcomes of pregnant women with GDM.

Objective:To explore the application value of case-based learning (CBL) and problem-based learning (PBL) dual-track teaching mode in the standardized resident training in laboratory medicine.Methods:The students of Grade 2017 and Grade 2018, who underwent standardized resident training of laboratory medicine in the Second Affiliated Hospital of Wenzhou Medical University from September 2017 to June 2021, were selected in this study. Seven students of Grade 2017 were served as the traditional lecture teaching group (traditional teaching group), and 12 students of Grade 2018 were assigned to the CBL-PBL dual-track teaching mode group (CBL-PBL group). Students of the two groups received 22 months of laboratory specialty training, and underwent one admission theory assessment,two mid-term theory assessments, annual professional level test, final theory assessment, final practical skills assessment as well as questionnaire survey. The questionnaire was distributed and finished anonymously after the final assessment. Survey indicators included ability assessment on solving clinical problem, assessment on the theoretical knowledge, computer operation skill, preparation time before teaching, teaching method satisfaction degree and clinical recognition. The results were divided into 5 categories: extremely agree, agree, general, disagree and extremely disagree, respectively (ranked as 5, 4, 3, 2, and 1). The Cronbach′s α coefficient was used to analyze the reliability of the questionnaire.Results:The mid-term theory assessment, annual professional level test and assessment results of clinical outcome interpretation were significantly higher in CBL-PBL dual-track teaching mode group than those in traditional lecture teaching group (all P<0.05). The performance after standardized resident training was significantly higher in the CBL-PBL group than in the traditional teaching group ( P<0.05). The Cronbach′s α coefficient of questionnaire reliability analysis was 0.938. Parameters assessment results including improved clinical problem-solving skills, computer operation skill, the ability to analyze and solve problems, the ability of innovation and adaptation, the learning interest, the ability of autonomous learning, teaching method satisfaction and conventional application to clinical recognition scores were all significantly better in the CBL-PBL group than those in the traditional teaching group (all P<0.05). Conclusion:The application of CBL-PBL dual-track teaching mode for the standardized resident training in laboratory medicine can effectively improve the assessment results and performance of students undergoing standardized resident training, and help to cultivate high quality medical laboratory professionals.

Objective To observe the value of isotropic volumetric MRI for displaying perineural spread(PNS)of cranial nerve(CN)in nasopharyngeal carcinoma.Methods Eighty-seven patients with pathologically proven nasopharyngeal carcinoma were prospectively enrolled.MR scanning,including three-dimensional liver acquisition with volume acceleration-flexible(3D LAVA_Flex)image,T2WI with fat suppression(T2WI-FS),T1WI,contrast enhancement(CE)T1WI-FS of nasopharynx and neck region were performed.The displaying rates of CN PNS were evaluated and compared between 3D LAVA Flex and T2WI-FS,T1WI,CE-T1WI-FS at patient level,CN group level and neural level,respectively.Results The displaying rate of CN PNS in all 87 nasopharyngeal carcinoma patients by 3D LAVA_Flex sequence was 49.43％(43/87),higher than that of conventional MRI(30/87,34.48％,P=0.001).Among 59 patients with advanced nasopharyngeal carcinoma diagnosed with conventional sequences,the displaying rate of CN PNS was 71.19％(42/59)by 3D LAVA-Flex sequence,higher than that of conventional MRI(30/59,50.85％,P=0.001).At both patient level and posterior CN level,significant differences of the displaying rate of CN PNS were found between 3D LAVA-Flex sequence and T2WI-FS,T1WI,CE-T1WI-FS,while at CN level,the displaying rates of mandibular nerve PNS,CN Ⅸ-Ⅺ PNS in jugular foramen(P＜0.05)and CN Ⅸ-Ⅻ PNS in carotid space of 3D LAVA_Flex sequence were all significantly higher than that of T2WI-FS,T1WI and CE-T1WI-FS(all P＜0.05),of PNS of CN Ⅲ-Ⅴ in cavernous sinus were higher than that of T2WI-FS(P＜0.05),while of PNS of hypoglossal nerve were significantly higher than that of T2WI-FS and T1WI(both P＜0.05).Conclusion 3D LAVA_Flex sequence could be used to effectively display CN PNS of nasopharyngeal carcinoma.

Objective To analyze the levels of IgE,TNF-α and FeNO in children with acute attack of bronchial asthma and their correlation with the severity of bronchial asthma, so as to provide theoretical basis for clinical evaluation of bronchial asthma. Methods A total of 547 children with acute bronchial asthma treated in Chengdu Women and Children's Central Hospital from January 2020 to December 2020 were selected and divided into mild group (n=287), moderate group (n=186) and severe group (n=74) according to the severity of their disease. All the children's symptoms were controlled after treatment. The serum IgE, TNF-α and FeNO levels in the experimental group were compared between the acute attack stage and the clinical control stage. Spearman correlation analysis was used to analyze the correlation between the serum IgE, TNF-α and FeNO levels and the severity of the disease. ROC curve of children with bronchial asthma was drawn to analyze the differential diagnosis value of serum IgE, TNF-α and FeNO levels in children with acute bronchial asthma. Results The levels of IgE, TNF-α and FeNO in acute stage were significantly higher than those in clinical control stage (P<0.05). The levels of serum IgE, TNF-α and FeNO in severe group were higher than those in mild and moderate groups significantly (P<0.05). The levels of serum IgE, TNF-α and FeNO in moderate group were higher than those in mild group significantly (P<0.05). Spearman correlation analysis showed that serum IgE, TNF-α and FeNO water were positively correlated with the severity of bronchial asthma (r=0.419 , 0.438 , 0.502 , P<0.05). ROC curve analysis showed that the AUC, sensitivity, accuracy and specificity of serum IgE, TNF-α and FeNO levels combined in diagnosing the severity of bronchial asthma in patients with acute attack was 0.938 (95% CI: 0.912-0.982 ), 83.47%, 92.06%, 94.28%. Conclusion The level of serum IgE, TNF-α and FeNO in children with acute attack of bronchial asthma is closely related to the severity of the disease, and combined detection of the three can be used to evaluate the severity of the disease in children.