Objective To study the role Erk/Slug signal pathway in the radiosensitivity of MDAMB-231 cells.Methods MDA-MB-231 cells were transfected with NF-κBp65 siRNA,PUMA siRNA and Slug siRNA respectively or treated with U0126 for 24h,then the cells were irradiated with 4 Gy γ-ray.At different time points post-irradiation,the expressions of Erk1/2,NF-κBp65,Slug and PUMA protein were detected.The cell survival rate and apoptotic index were detected by MTT and TUNEL methods.Resluts Compared with 4 Gy irradiated group,the expression of PUMA was reduced in NF-κB p65 siRNA/4 Gy group,the expressions of Slug and Erk1/2 were obviously decreased but PUMA increased in U0126/4 Gy group,the expression of Erk1/2 had no change but the expression of PUMA increased significantly in Slug siRNA/4 Gyγ group.Meanwhile,at 48h post-irradiation,for U0126/4 Gy group and Slug siRNA/4 Gy group,cells survival rates were decreased to 19.78 ±2.71 (F=11.39,P＜0.05) and 17.41 ±4.58(F=15.31,P＜0.05),cell apoptosis rates were 28.61 ±4.70 (F=9.84,P＜0.05) and 27.55±6.41(F =10.31,P ＜ 0.05),respectively.At 24 h post-irradiation,for NF-κB p65 siRNA/4 Gy group and PUMA siRNA/4 Gy group,cell survival rates approached to 85.65 ± 9.60 (F =12.31,P ＜ 0.05) and 87.53±11.50 (F=13.68,P＜0.05),and cell apoptosis rates declined to 3.28 ±0.78 (F=10.83,P ＜ 0.05) and 3.46 ± 0.84 (F =9.92,P ＜ 0.05).Conclusions The radiosensitivity of MDA-MB-231cells was relative to the induction of NF-κB up-regulated PUMA,and the radioresistance was caused by the up-regulation of Slug induced by Erk1/2,which inhibited the expression of PUMA.

Purpose To study the clinical features,pathological manifestation and immunohistochemical phenotype and improve the diagnosis and treatment of myoepithelial carcinoma in salivary glands.Methods Histomorphology and immunohistochemical phenotype were analyzed after the sections were stained with routine HE and immunohistochemical methods,and the relevant literatures were reviewed.Results The tumours were predominantly composed of pale-stained clear cells.In some cases,plasma-like cells,epithelioid cells and spindle cells were also seen.The cells were arranged in nest,solid or cords.Mitosis was easily seen,cytological atypia was obvious and necrosis existed in 4 cases.The results of immunohistochemical staining showed that CK was expressed in all cases.EMA was expressed in 8 cases.p63 and CK5/6 were expressed in 11 cases.S-100 was expressed in 10 cases.vimentin was expressed in 4 cases.Calponin was expressed in 2 cases.SMA was expressed in one case.The proliferation index of Ki-67 was 5％ to 40％.Conclusion The histological changes of myoepithelial carcinoma cells are diverse,and pathological and immunohistochemical methods are helpful for improving the rate of right diagnosis.Sugery is the main treatment for myoepithelial carcinoma.

Objective To investigate the effect of anti-proliferation on pioglitazone of human gastric cancer SGC-7901 cells. Methods Different concentrations of PGZ groups (0.01,0.1,1.0,10.0,50.0 and 100.0 mol/L) were set up, con-trol group were without drugs. The anti-proliferation effect of PGZ on human gastric cancer SGC-7901 cells were de-tected by MTT method. Results Compared with the control group,the OD value of gastric cancer SGC-7901 cells through different concentrations PGZ(0.1,1.0,10.0,50.0 and 100.0 mol/L) incubating 12,24,48,72 h were higher,the differences were significant (P<0.05). The concentration of 0.01 mol/L PGZ had no significant inhibition on the prolif-eration of gastric cancer SGC-7901 cells(P>0.05),the other dose groups had significant effect of inhibition on the pro-liferation of gastric cancer SGC-7901 cells,compared with the control group,the differences were significant (P<0.05). Conclusion The PGZ is capable of inhibiting proliferation of human gastric cancer SGC-7901 cells in vitro significant-ly, and the result of inhibiting proliferation rate shows a significant dose-time relationship.

Objective To explore the molecular mechanism of Atractylodes macrocephala in the treatment of Ulcerative colitis(UC)based on network pharmacology,and verify it with animal experiments.Methods The active components of Atractylodes macrocephala was screened from the TCMSP database,the TCM-ID database,and in combination with relevant references,and the corresponding targets were obtained through Swiss database.The relevant targets of UC were obtained from GeneCards database,construct the"drug-component-target-disease"network diagram and"pathway-active ingredient-target"network diagram and draw PPI network diagram;GO function enrichment analysis and KEGG signal pathway annotation analysis were carried out.Autodock software is used for molecular docking of active components and targets.Then,the experimental validation of the network pharmacology prediction was carried out.The mouse UC model was induced by dextran sodium sulfate(DSS).The pathological changes of the colon tissue,the number of goblet cells,and the positive expression of inflammatory factorswere detected by HE staining,AB-PAS staining and immunohistochemistry in colon tissue of UC mice.Results The results have shown 30 active ingredients including atractylolactone I,II and III were screened,and 591 corresponding targets were obtained,of which the key target was IL-1β、TNF-α and so on.Molecular docking show that the core components had good binding affinity with the key targets.And the results of animal experiments showed that the alcohol extract of Atractylodes macrocephala could significantly increase the colon length,reduce the DAI score,improve the pathological changes of colon tissue of UC mice,increase the number of goblet cells,and inhibit the expression of IL-1β,TNF-α in colon tissue.Conclusion This study indicated that Atractylodes macrocephala could regulate the release of inflammatory factors through multiple components,multi-target and multi-channel,which could inhibit inflammatory reaction and play a role in improving UC.

Objective:To summarize the case data of endometrial cancer (EC), analyze the related factors of lymph node metastasis, and establish the prediction model, so as to provide reference for clinical practice.Methods:191 patients with endometrial cancer who were diagnosed and treated in department of gynecology of Baoding Maternal and Child Health Hospital from January 2010 to December 2019 were selected as the research objects. The demographic and surgical pathological information of the patients were analyzed retrospectively. The risk factors of lymph node metastasis were analyzed by univariate and logistic regression analysis, and the predictive model was established.Results:A total of 191 patients with EC, aged 26-76(53.1±9.5)years old, body mass index (BMI)18.70-40.20(25.84±3.94)kg/m 2, 13 cases (6. 81%) had lymph node metastasis. Univariate analysis showed that lymph node metastasis was associated with obesity (BMI≥28 kg/m 2), pathological type (non endometrioid adenocarcinoma), degree of differentiation, depth of myometrial invasion (>1/2) and vascular invasion ( P<0.05). Logistic multivariate analysis showed that low differentiation ( OR=9.475, 95% CI: 1.840-48.799), vascular invasion ( OR=6.614, 95% CI: 1.457-30.024) and deep muscle invasion ( OR=4.997, 95% CI: 1.342-18.600) were independent risk factors ( P<0.05). The regression equation: Logit P=-4.488+ 1.609× myometrial infiltration depth+ 1.889×vascular infiltration+ 2.249×degree of tissue differentiation. The area under the receiver operating characteristic (ROC) curve (AUC) of EC lymph node metastasis probability P was 0.813 (95% CI: 0.688-0.938). The cut off value of 0.56 was ideal. At this time, the prediction sensitivity was 76.9% and the specificity was 79.2%. Conclusions:In clinical practice, gynecologists should consider the condition of EC patients and make operation plan to avoid over treatment or under treatment.

Objective: To investigate the effects of hesperetin on fine particulate matter (PM_2.5) induced apoptosis in H9c2 cells and related mechanisms. Methods: H9c2 cells were divided into 4 groups: control group (cells were cultured without intervention), PM_2.5 group (cells were treated with 800 µg/ml PM_2.5), hesperetin group (H group, cells were treated by 40 µmol/L hesperetin for 1 h at 37 ℃), and hesperetin+PM_2.5 group (H+PM_2.5 group, cells were pretreated with hesperetin before PM_2.5 treatment). Cells were cultured for corresponding interval. Apoptotic cells were detected by Annexin Ⅴ-FITC/PI apoptosis detection kit and Hoechst staining. The intracellular reactive oxygen species (ROS) levels were measured by DCFH-DA Fluorescence Probe and mitochondrial membrane potential (MMP) was detected with JC-1 staining, respectively in these groups. Apoptotic related protein and phosphorylated MAPK expression levels were determined by Western blot. Results: (1) Flow cytometry results showed that the apoptosis rate of PM_2.5 group ((48.94±3.20)%) was significantly higher than that of control group ((8.13±1.40)%, P<0.01), which was significantly reduced in H+PM_2.5 group ((34.80±2.21)%) (P=0.003 2 vs. PM_2.5 group, P<0.01 vs. control group). The number of Hoechst 33258 positive apoptotic cells was distinctly less in H+PM_2.5 group than in PM_2.5 group. (2) The ROS levels was significantly higher in PM_2.5 group ((49.69±5.05)%) than in control group (10.57±1.33)%, P<0.01), which was significantly reduced in H+PM_2.5 group ((35.08±3.90)%) (P=0.000 2 vs. PM_2.5 group, P<0.01 vs. control group). (3) Green fluorescence indicating the JC-1 monomer form, which represented MMP loss of H9c2 cells, was significantly higher in PM_2.5 group ((20.28±4.69)%) than in control group ((10.50±2.72)%, P<0.01), which was significantly decreased in H+PM_2.5 group ((13.41±2.89)%) (P<0.01 vs. PM_2.5 group, P=0.029 4 vs. control group). (4) The expression levels of Bcl-2 protein of H9c2 cells was lower in PM_2.5 group ((76.94±4.52)%) than in control group (100%, P=0.000 9), which was significantly upregulated in H+PM_2.5 group ((92.95±6.82)%) (P=0.027 5 vs. PM_2.5 group, P=0.15 vs. control group). The expression levels of cleaved caspase-3 protein of H9c2 cells was significantly higher in PM₂.5 group ((243.98±17.94)%) than in control group (100%, P=0.000 2), which was significantly downregulated in H+PM_2.5 group ((200.45±4.31)%) (P=0.015 vs. PM_2.5 group, P<0.01 vs. control group). (5) The expression levels of phosphorylated p38 MAPK protein of H9c2 cells was higher in PM_2.5 group ((118.90±4.78)%) than in control group(100%, P=0.002 7), which could be significantly downregulated in H+PM_2.5 group ((103.30±1.27)%) (P=0.01 vs. PM_2.5 group, P=0.05 vs. control group). The expression levels of phosphorylated ERK protein of H9c2 cells was higher in PM_2.5 group ((163.50±4.98)%) than in control group (100%, P<0.01), which was significantly downregulated in H+PM_2.5 group ((139.10±2.72)%) (P=0.001 6 vs. PM_2.5 group, P<0.01 vs. control group). Conclusions Hesperetin protects H9c2 cells from PM_2.5 stimulation through reducing oxidative stress and protecting mitochondrial function, regulating the expression of apoptotic associated proteins as well as MAPK signal pathway, thus inhibiting H9c2 cells apoptosis.

Objective:To investigate the effects and significance of acute and chronic trauma on brain degree centrality (DC) in patients with post-traumatic stress disorder (PTSD) who lost their only child at resting state.Methods:Retrospectively, the study enrolled a total of 51 parents with PTSD, including 35 PTSD parents whose children was lost in emergencies (acute bereaved PTSD group) and 16 PTSD parents whose children was lost of chronic causes such as diseases (chronic bereaved PTSD group). Fifty local adults were also included as healthy controls (HC group). The clinical administered PTSD scale(CAPS) was used to evaluate the severity of the subjects' clinical symptoms.Resting-state functional magnetic resonance imaging(fMRI) data of all subjects were collected and DC values were calculated.SPSS 22.0 software was used for statistical analysis.Covariance analysis was performed among three groups, while post hoc was performed between any two groups.What's more, correlation analyses were utilized between abnormal brain regions and the scores of CAPS.Results:Significant group effects were found in multiple regions, including the right inferior temporal gyrus (MNI: x, y, z=66, -27, -21), right temporal pole (MNI: x, y, z=54, 15, -9), right orbital inferior frontal gyrus (MNI: x, y, z=42, 21, -15), bilateral medial superior frontal gyri (MNI: right x, y, z=6, 63, 12; left x, y, z=-3, 60, 18), left inferior parietal angular gyrus (MNI: x, y, z=-45, -36, 51) and left postcentral gyrus (MNI: x, y, z=-45, -33, 51). Compared with HC group, the DC of two patient groups increased in the right inferior temporal gyrus (MNI: acute x, y, z=63, -27, -21; chronic x, y, z=63, -21, -27); the DC of acute bereaved PTSD group decreased in the right temporal pole (MNI: x, y, z=45, 21, -15) and the right orbital inferior frontal gyrus (MNI: x, y, z=48, 24, -12), while the DC of chronic bereaved PTSD group decreased in the left inferior parietal angular gyrus (MNI: x, y, z=-45, -36, 51) and left postcentral gyrus (MNI: x, y, z=-45, -33, 51). Compared with chronic bereaved PTSD group, the DC of acute bereaved PTSD group increased in the left inferior parietal angular gyrus (MNI: x, y, z=-33, -39, 42) and left postcentral gyrus (MNI: x, y, z=-45, -33, 51), while decreased in the right temporal pole (MNI: x, y, z=51, 12, -9), right orbital inferior frontal gyrus (MNI: x, y, z=42, 21, -15) and bilateral medial superior frontal gyri (MNI: left x, y, z=0, 57, 15; right x, y, z=3, 57, 15). In chronic bereaved PTSD group, the DC of the left postcentral gyrus was negatively correlated with C1 (avoid trauma-related thoughts, feelings) score in CAPS ( r=-0.606, P=0.028). In acute bereaved PTSD group, the DC of the left medial superior frontal gyrus was negatively correlated with D4 (high vigilance) score ( r=-0.416, P=0.020). Conclusion:There exist functional abnormalities of multiple brain regions in acute and chronic bereaved parents with PTSD.The high arousal symptoms of the former may be related with the abnormalities of prefrontal-amygdala neural circuit, while the latter show higher avoidance which may be associated with the dysfunction of somatosensory brain regions such as postcentral gyrus.

Objective:To investigate the effect of coping style on the gray matter volume in patients with post-traumatic stress disorder (PTSD) who lost their only child, and the mediating role of gray matter volume in evaluating the influence of coping style in clinical symptoms of these parents.Methods:A total of 57 parents with PTSD (PTSD group) and 162 parents without PTSD (non-PTSD group) who lost their only child from September 2016 to March 2017 were enrolled from Jiangsu Province, China. Brain MRI data at resting state were collected. Voxel-based multiple regression analysis was performed to confirm the brain areas in which coping style main effect, diagnosis main effect and their interaction had significant influences in gray matter volumes. Correlations among gray matter volume of brain areas related to coping style, coping style scale scores, and clinician-administered PTSD scale (CAPS) scores were analyzed. Structural equation modeling was used to analyze the mediating role of gray matter volume in the influence of coping style in clinical symptoms of parents lost their only child.Results:(1) The coping style main effect did not significantly influence the gray matter volume in all subjects, and the diagnosis main effect had significant influence in gray matter volume in the right lingual gyrus; their interaction had significant influence in gray matter volume in the right peritalar fissure cortex and lingual gyrus. The positive coping style in the PTSD group had significant influence in the gray matter volumes of the right peritalar fissure cortex and lingual gyrus. (2) In the PTSD group, the scores of positive coping style were positively correlated with the gray matter volumes of the right talus fissure and the lingual gyrus ( P<0.05); the scores of positive coping style, and the gray matter volumes of the right talus fissure and the lingual gyrus were negatively correlated with scores of CAPS-C 5 and CAPS-C ( P<0.05). (3) In the PTSD group, positive coping style can positively predict the gray matter volumes of the right talus fissure and the lingual gyrus; the gray matter volumes of the right talus fissure and the lingual gyrus can negatively predict the avoidance-related symptoms. Conclusion:Positive coping style has influence in the gray matter volumes of the right talar fissure and lingual gyrus of PTSD patients lost their only child; and less positive coping style may affect the brain areas related to visual information processing, thus aggravating avoidance-related symptoms of PTSD patients.

OBJECTIVE: To investigate the effect of orthodontic traction on the microstructure of dental enamel. METHODS: Forty-eight isolated premolars were randomly divided into 6 groups (=8), including Group A (blank control group), in which the teeth were bonded with the orthodontic brackets without any loading force; Groups B1, B2, and B3 where the teeth were bonded with the orthodontic brackets using clinical adhesives and loaded with 50 g force for 6 months, 200 g force for 6 months, and 200 g force for 1 month, respectively; and Groups C1 and C2, where the teeth were bonded with straight wire brackets using light curing bonding and chemical curing bonding techniques, respectively. All the teeth were embedded with non-decalcified epoxy resin. Scanning electron microscope (SEM), atomic force microscope (AFM), and energy spectrometer (EDS) were used to analyze interface morphology and elemental composition of the teeth sliced with a hard tissue microtome. RESULTS: Compared with those in Group A, the teeth in the other 5 groups showed increased adhesive residue index with microcracks and void structures on the enamel surface under SEM; AFM revealed microcracks on the enamel surface with angles to the grinding direction. A larger loading force on the bracket resulted in more microcracks on the enamel interface. The interface roughness differed significantly between Groups A and C2, and the peak-to-valley distance differed significantly between Groups A, C, and C2. CONCLUSIONS Orthodontic traction can cause changes in the microstructure of normal dental enamel.
Dental Enamel ; Materials Testing ; Orthodontic Brackets ; Resin Cements ; Surface Properties ; Traction

OBJECTIVETo evaluate the application of mismatch repair (MMR) genes proteins expression to screen for Lynch syndrome in colorectal cancer patients.

METHODSOne hundred consecutive colorectal cancers cases collected from 2012 to 2013 were tested immunohistochemically for the protein expression of MLH1, MSH2, MSH6 and PMS2, and also by the ARMS method for the mutation status of BRAF genes in those cases lacking protein expression for MLH1.

RESULTSThe result of MMR immunocytochemistry showed that nine of 100 cases lacked MMR protein expression, including three cases each that were MLH1-/PMS2- and MSH2-/MSH6- respectively, two cases were MLH6- and one case was PMS2-; overall, the majority of these cases lacked protein expression of MLH1 and MSH2. The BRAF genes mutation test showed one case of mutation, indicating that the patient might have MLH1 gene methylation as a result of the mutation of BRAF genes, and that was a sporadic case. The age of onset for patients lacking MMR protein expression was lower than patients with sporadic colorectal cancer (P = 0.011). Colorectal cancers associated with the lack of MMR protein expression mostly occurred in the right colon (P = 0.001), and histomorphologically were often accompanied by mucinous adenocarcinoma (P = 0.010) and tumor lymphocytic infiltration.

CONCLUSIONImmunohistochemical staining for MMR proteins in patients with colorectal cancer, accompanied by testing for BRAF genes mutation, may be an effective approach to screen for Lynch syndrome.

Adaptor Proteins, Signal Transducing ; genetics ; metabolism ; Colorectal Neoplasms, Hereditary Nonpolyposis ; diagnosis ; genetics ; DNA Mismatch Repair ; Humans ; Immunohistochemistry ; MutL Protein Homolog 1 ; Mutation ; Nuclear Proteins ; genetics ; metabolism ; Proto-Oncogene Proteins B-raf ; genetics ; metabolism