Objective:To study the effects of exosomes (EXO) released by adenoid cystic carcinoma SACC-83 cells on the expression of fibroblast activation protein (FAP) in normal human salivary gland stromal fibroblasts (hSGSFs).Methods:ACC exosomes were isolated from SACC-83 cell culture supernatant by using Total Exosome Isolation Reagent.The whole-mount EXO were characterized and assessed by transmission electron microscope and Western Blot.The exosomes were labeled with green fluorescent dye PKH67 and co-cultured with hSGSFs for 48h,followed by staining with Alexa Fluor 594 Phalloidin and DAPI.Mterwards,exsosomes uptake was observed under a laser scanning confocal microscope.After a 48-hour co-culture of SACC-83 exosomes with hSGSFs,the expression of FAP in SACC-83-EXO-treated hSGSFs was investigated by qRT-PCR and Western Blot.Results:The vesicles isolated from SACC-83 cell culture supernatant had the reported size range of 30-100 nm,expressed the exosomal marker CD63 and TSG101.Mter co-culture of hSGSFs with PKH67 labeled SACC-83 exosomes,exosomes were taken up by hSGSFs and FAP expression was elevated in hSGSFs.Conclusion:Exosomes derived from SACC-83 cells can be taken up by hSGSFs and can induce the expression of FAP in hSGSFs.These results suggest that exosomes derived from SACC-83 cells might induce the transformation of normal salivary gland strormal fibroblasts to cancer associated fibroblasts.

To explore the status and characteristics of clinical trials of therapeutic cancer vaccines, and provide the overall trend of clinical translational research of therapeutic cancer vaccines.

Objective To investigate the effects of circadian rhythm on interictal epileptiform discharges in patients with localization-related epilepsy.Methods Patients diagnosed with epilepsy in Sanbo Brain Hospital from January 2011 to January 2012 participated in this study.All patients were subjected to comprehensive evaluation,which included prolonged video-electroencephalogram (EEG),magnetic resonance imaging.Intracranial electrodes,PET,SPECT were also adopted if necessary.Circadian rhythm was divided into four stages:REM,NREM Ⅰ-Ⅱ,NREM Ⅲ-Ⅳ,and waking.The amount and distribution of ⅡD were compared by ANOVA.Results Significant differences in the amount and distribution of ⅡD were found among NREM Ⅰ-Ⅱ,NREM Ⅲ-Ⅳ,REM,and waking.However,no differences in the amount and distribution of ⅡD were noted between NREM Ⅰ-Ⅱ and NREM Ⅲ-Ⅳ as well as between REM and waking.Conclusion The amount of ⅡD is higher in NREM than in REM and waking;thus,NREM is more sensitive to diagnose epilepsy.The distribution of ⅡD in REM and waking is more restricted than that in NREM.

Aim To investigate the effect of p38 MAPK AS-ODNs on the expression of GLT-1 during the induction of brain ischemic tolerance induced by cerebral ischemic preconditioning (CIP).Methods Eighty healthy male Wistar rats with permanent occlusion of the bilateral vertebral arteries were randomly divided into 6 groups: ①Sham group (n=10);②CIP group (n=10);③ischemic insult (Ⅱ) group (n=10);④CIP+Ⅱ group (n=10);⑤p38 MAPK AS-ODNs+CIP+Ⅱ group (n=30);⑥p38 MAPK S-ODNs+CIP+Ⅱ group (n=10).Group ⑤ was divided into 5 nmol, 10 nmol and 15 nmol subgroups according to the dose of p38 MAPK AS-ODNs (n=10).The dose of p38 MAPK S-ODNs was 15 nmol.All the rats were sacrificed 6 h and 2 d after the sham operation or the last time of global brain ischemia reperfusion.Western blot and immunohistochemistry analysis were used for detecting the expression of p-p38 MAPK and GLT-1 protein.Results CIP moderately up-regulated the expression of p-p38 MAPK and significantly up-regulated the expression of GLT-1 protein, inhibited the excessively up-regulation of p-p38 MAPK and the down-regulation of GLT-1 induced by ischemic insult.p38 MAPK AS-ODNs significantly inhibited the up-regulation of p-p38 MAPK and GLT-1 protein in a dose-dependent manner during the induction of brain ischemic tolerance by CIP.Conclusion p38 MAPK AS-ODNs inhibit the up-regulation of GLT-1 during the induction of brain ischemic tolerance induced by CIP.

OBJECTIVE:To establish a method for the simultaneous determination of paeoniflorin and paeonol in Dirda pill, and provide reference for improving its quality control standard. METHODS:HPLC was performed on the column of ZORBAX C18 with mobile phase of acetonitrile-0.1% phosphoric acid (gradient elutio) at a flow rate of 0.8 ml/min,detection wavelength was 230 nm(paeoniflorin),274 nm(paeonol),columne temperature was 35 ℃,injection volume was 20 μl. RESULTS:The linear range was 0.514 8-1.544 5μg for paeoniflorin(r=0.999 2)and 0.643 2-1.960 4μg for paeonol(r=0.999 8);the limits of quantifi-cation were 0.135 6,0.126 4 μg,limits of detection were 0.067 8,0.063 2 μg;RSDs of precision,stability and reproducibility tests were lower than 2%;recoveries were 95.78%-97.27%(RSD=0.62%,n=6) and 97.38%-98.38%(RSD=0.42%,n=6). CONCLUSIONS:The method is simple with good reprosucibility and high sensibility,and can be used for the simultaneous deter-mination of paeoniflorin and paeonol in Dirda pill.

Objective: To establish a nomograph model for prediction of cervical central lymph node metastasis (CLNM) among patients with thyroid papillary carcinoma (PTC), so as to provide the evidence for designing personalized treatment plans for PTC. Methods : The data of patients that underwent thyroidectomy and were pathologically diagnosed with PTC post-surgery in the Affiliated Traditional Chinese Medicine Hospital of Xinjiang Medical University from 2018 to 2021 were collected. Patients' data captured from 2018 to 2020 and from 2021 were used as the training set and the validation set, respectively. Predictive factors were screened using a multivariable logistic regression model, and the nomograph model for prediction of CLNM risk was established. The predictive value of the model was evaluated using the receiver operating characteristic (ROC) curve and the adjusted curve. Results: Totally 1 820 PTC cases were included in the training set, including 458 cases with CLNM (25.16%), and 797 cases in the validation set, including 207 cases with CLNM (25.98%). The prediction model is p=ey/(1+ey), y=0.761 + 0.525 × sex + (-0.039) ×age + 0.351 × extrathyroid invasion + 0.368 × neck lymph node enlargement + 1.021×maximum tumor diameter + (-0.009) × TT4 + (-0.001) × anti-TPOAb. The area under the ROC curve was 0.732 for the training set and 0.731 for the validation set, and Hosmer-Lemeshow test showed a good fitting effect (P=0.936, 0.722). Conclusion The nomograph model constructed in this study has a high predictive value for CLNM among patients with PTC.

Objective To compare the classification rate of three classifications of epilepsy syndromes proposed by International League Against Epilepsy(ILAE),and analyze their stuctural changes.Methods All patients with epilepsy who consecutively visited the epilepsy center of Peking Union Medical College Hospital between Aug.1st,2007 and Mat.31st.2008 were included.Thtee classifications of epilepsy syndromes were used in order.Results In this study,we could categorize 75.5 % of 1356 patients by applying the 1989 international classification of epilepsy syndromes.89.0 % of them by the 2001 proposed diagnostic scheme and 88.1 % of them by the 2006 report.In this aspect,the 2001 and 2006 classifications were better than the 1989 classification(x2=116.3,P<0.01).However,only 11.6 % (157),12.O % (162)and 11.9 % (160)of patients with specific epilepsy syndromes were identified from the 1356 epileptic patients by three classifications.respectively.This data based on the 2001 and 2006 classifications did not change markedly in comparison with the 1989 classification(x2=0.09,P>0.05).Conclusions The 2006 report involve mole scientific mode of classification and systematic evaluation,and can classify more patients with epilepsy.It can be ased in clinical and scientific research.which can not only accumulate data for developing more scientific classification but also stimulate research especially in the fields of genetics and functional morphology.

Objective To investigate the clinical,electroencephalogram (EEG) and genetic features of nocturnal frontal lobe epilepsy (NFLE) in the Chinese population.Methods Clinical examination,EEG recording,mutation screenings in transmembrane domains 1-3 of neuronal nicotinic acetylcholine receptor (nAChR) α4 (CHRNA4),β2 (CHRNB2) and α2 (CHRNA2) using PCR amplification and sequencing were carried out on 6 patients and some members in 3 families with NFLE.Results Among 6 patients (5 male) with NFLE,the mean age was (20.5±11.5) years and the mean age at onset was (7.3±5.5) years.Clinical features included seizures of dystonic posturing in 2 patients and seizures of hyperkinetic movements in 4 patients with the maximum frequency of 6 seizures within one night.The ictal and interictal video-EEG (VEEG) of frontal lobes showed epileptic discharges,slow wave activity,normal activity or electrode artifacts.There weren' t abnormity in other clinical examination and neuroimagings.No mutations were identified in the genes screened.Conclusion NFLE is a heterogenetic epilepsy syndrome.

Objective To observe the expression of indoleamine 2,3-dioxy-genase(IDO) in hippocampus of rats with posttraumatic stress disorder (PTSD) and the protective effect of IDO inhibitor on neurons,and to explore the role of IDO in the pathogenesis of PTSD.Methods Adult male Wistar rats were randomly divided into the normal control group,PTSD model group and IDO inhibitor treatment group.The expression of IDO was detected by immunohistochemistry,RT-PCR and Western-blot.The apoptosis of rat hippocampal neurons was assayed by Tunel staining.Tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) were detected by ELISA.Moreover behavioral evaluation was performed,including central residence time,percentage of open arm residence time and stage latency.Results Comparing with the control group,PTSD rats showed decreased central residence time ((22.65± 1.54)s),decreased percentage of open arm residence time((10.55± 1.96) ％),prolonged stage latency ((56.38±4.21) s) (P＜0.05),increased TNF-α ((8.58±0.6) pg/ml),IL-6 ((15.72±1.42) pg/ml) and IDO mRNA (0.8278±0.0796),increased IDO protein (1.2329±0.1148) expression and apoptosis rate ((81.47± 6.86) ％) in hippocampus (P＜ 0.05) (P＜ 0.05).However,rats treated with IDO inhibitor showed increased central residence time((30.78±3.20) s),increased percentage of open arm residence time ((10.55± 1.96)％),shortened stage latency ((56.38 4.21) s),meanwhile reduced expression of TNF-α((3.69±0.41) pg/ml),IL-6((7.45±0.58) pg/ml),IDO mRNA(0.2236 ±0.0387) and IDO protein(0.4235±0.0411) was detected in hippocampus(P＜0.05).Apoptosis rate ((42.54± 3.98)％) was also decreased in hippocampus(P＜0.05).Conclusion The content of TNF-α,IL-6 and IDO are increased significantly in the hippocampus of PTSD rats.IDO may participate in the pathogenesis of PTSD,and the IDO inhibitor may play a neuroprotective role in hippocampus of PTSD.

Objective To investigate the effects of miR?23a?3p on proliferation, migration and apoptosis on human acute myeloid leukemia ( AML) cells by targeting SMC1A. Methods Microarray analysis was used to screen differentially expressed microRNAs and mRNAs in human AML cells. Real?time fluorescence quantitative PCR (RT?qRCR) was used to detect the expressions of miR?23a?3p and SMCA in human AML cell line U937. TargetScan database was used to analyze the correlation between miR?23a?3p and SMC1A. Double luciferase reporter gene was used to detect the interaction between miR?23a?3p and SMC1A. The effect of miR?23a?3p expression on the proliferation of U937 cells was detected by clonal assay. The migration, apoptosis, cell cycle and caspase?3 activity of U937 cells regulated by miR?23a?3p were detected by cell scratch assay and flow cytometry, respectively. Western blot was used to detect the expressions of Bax and Bcl?2 in U937 cells. Results Compared with human normal monocyte SC group (1.00), the expression of miR?23a?3p in U937 cells was up?regulated (2.56±0.78) ( P＜0.01), while the expression of SMC1A was down?regulated (0.48±0.56, P＜0.01). miR?23a?3p specifically bond to SMC1A 3′UTR and regulated the expression activity of SMC1A. Overexpression of miR?23a?3p promoted the proliferation and migration of U937 cells and inhibited the apoptosis of U937 cells, while up?regulation of SMC1A inhibited the proliferation and migration of U937 cells and promoted the apoptosis of U937 cells. The percentages of G0／G1 phase, G2／M phase and S phase cells in the negative control group were ( 37.48 ± 0.21)%, (16.78±0.18)% and (45.74±0.15)% respectively, and those in the miR?23a?3p mimics group were (19.96±0.11)%, (41.69±0.24)% and (38.24±0.34)%, respectively. The difference was statistically significant (all P＜0.05). The proportions of G0／G1 phase, G2／M phase and S phase cells in the group of miR?23a?3p mimics+pcDNA3.1?SMC1A were (36.88± 0.21)%, ( 30.44± 0.33)% and ( 32.88± 0.16)%, respectively, without significant difference when compared with those of the miR?23a?3p mimics group ( P＞0.05). The relative expression levels of Bax and Bcl?2 protein in the negative control group were 0.55±0.45 and 0.31±0.54, respectively. Overexpression of miR?23a?3p inhibited the expression of Bax protein in U937 cells (0.23± 0.13, P＜0.001), promoted the expression of Bcl?2 protein ( 0.50 ± 0.23, P＜0.01), while SMC1A increased the expression of Bax protein in U937 cells (0.40± 0.11, P＜0.01), and inhibited the expression of Bcl?2 protein (0.37± 0.15). In the negative control group, caspase?3 activity was (25.82± 0.89)%.Overexpression of miR?23a?3p inhibited caspase?3 activity in U937 cells (3.64±0.56)%, P＜0.01, while up?regulation of SMC1A promoted caspase?3 activity in U937 cells ( 15.29 ± 0.85)%, P＜0.01. Conclusion miR?23a?3p can inhibit the proliferation and migration and promote apoptosis of human AML cells by targeting SMC1A.