Background and purpose:PTEN mutation has been found in 20%-40% of malignant gliomas.The common mutant epidermal growth factor receptor(EGFR vⅢ)was reported to coexpress in PTEN-deficient EGFR-expressing tumor.PTEN has been shown to interact directly with FAK and reduce its tyrosine phosphorylation levels to inhibit cell invasion.The invasion of glioma cells with EGFRvⅢ expression and PTEN deficiency is increased.This study was to observe whether PTEN inhibits glioma cell invasion even in the presence of strong pro-invasive signals provided by constitutive EGFR activity.Methods:U87?EGFR cells were transfected with pcDNA3.1 constructs encoding PTEN and the cells invasion levels were detected by transwell invasion assay.The expression of FAK was detected by immunoblotting.FAK expression vector was transfected into U87?EGFR-wtPTEN cells and the change of cells invasion was documented.Results:PTEN and PTEN(G129E)could inhibit cell invasion induced by EGFRvⅢ.PTEN and PTEN(G129E)could decrease the FAK phosphorylation at Tyr397.Over expression of FAK in U87?EGFR-PTEN abrogated PTEN-induced down-regulation of the phosphorylation status of FAK and rescued cell invasion.Conclusions:PTEN could inhibit cell invasion induced by EGFRvⅢ by dephosphorylating FAK.

Objective To investigate the association between the functional polymorphisms of Foxp3 gene and unexplained recurrent spontaneous abortion (URSA).Methods PCR-restriction fragment length polymorphism (rs3761548,rs2294021 ) and PCR with sequence-specific primers (rs2232365,rs5902434) were used to detect four polymorphisms of Foxp3 in 146 URSA cases and 112 normal controls.Results ( 1 ) The frequencies of rs3761548A/C were 10.3％,22.3％ in genotype C/C,38.4％,40.2％ in genotype A/C and 51.4％,37.5％ in genotype A/A between URSA patients and normal controls; the frequencies of rs2232365A/G were 5.5％,15.2％ in genotype A/A,47.9％,50.0％ in genotype A/G,46.6％,34.8％ in genotype G/G between URSA patients and normal controls; they all reached statistical difference ( P＜0.05 ).The carriers of rs3761548A allele and rs2232365G allele increased the risk of URSA (OR=1.73,1.61 ; all P ＜ 0.05 ).(2) There was no difference in the genotypic distribution of rs5902434del/ArTTpolymorphism between cases and controls ( P =0.10),but the frequency of del allele in URSA was statistically increased than that of controls (71.2％,62.5％ ; OR =1.49,P =0.04 ).(3) There was no different distribution in 3 genotypes (C/C,T/C,T/T) and 2 alleles (T and C) of rs2294021T/C between URSA patients and normal controls (P =0.18 and 0.08 ).(4) Estimated haplotype frequency distribution of rs5902434del/ATT,rs3761548A/C and rs22323565A/G showed haplotype del-A-G conferring the susceptibility to URSA ( OR =2.51,P ＜ 0.01 ) but haplotype del-C-G and ATT-A-A could provide protection on URSA ( OR =0.18,0.22 ; all P ＜ 0.01 ).Conclusion Functional polymorphisms of Foxp3 gene could probably confer the susceptibility to URSA,by altering Foxp3 function and (or) its expression.

Objective To compare the application and clinical significance of the liquid based cytology examination and the DNA quantitative analysis in female cervical lesions.Methods The cervical cell samples were collected from 879 women participating in the comparison by the cervical brush and performed the the liquid-based thin layer section preparation for conducting Papanicolaou staining and DNA staining respectively.The liquid based cytology examination was performed on the Papanicolaou staining section and the fully automatic scanning diagnosis was performed on the DNA staining section.Results The cases of above atypical squa-mous cells of undetermined significance(ASCUS)detected by the liquid based cytology examination and the partial cases of hetero-ploid cell detected by the fully automated DNA ploidy analysis system were recommended to further perform colposcopy and cervi-cal biopsy.28 women were performed the pathological biopsy.With the cytological examination result as the standard,the detection rate of above ASCUS cervical lesions detected by the cellular DNA quantitative analysis was calculated.Conclusion The combined application of the cellular DNA quantitative analysis method and the liquid based cytology examination can obviously increase the positive detection rate of cervical cancer and precancerous lesion,which has important significance for the prevention and treatment of female cervical cancer in our country.

Dimerization among the EGFR family of tyrosine kinase receptors leads to allosteric activation of the kinase domains of the partners. Unlike other members in the family, the kinase domain of HER3 lacks key amino acid residues for catalytic activity. As a result, HER3 is suggested to serve as an allosteric activator of other EGFR family members which include EGFR, HER2 and HER4. To study the role of intracellular domains in HER3 dimerization and activation of downstream signaling pathways, we constructed HER3/HER2 chimeric receptors by replacing the HER3 kinase domain (HER3-2-3) or both the kinase domain and the C-terminal tail (HER3-2-2) with the HER2 counterparts and expressed the chimeric receptors in Chinese hamster ovary (CHO) cells. While over expression of the intact human HER3 transformed CHO cells with oncogenic properties such as AKT/ERK activation and increased proliferation and migration, CHO cells expressing the HER3-2-3 chimeric receptor showed significantly reduced HER3/HER2 dimerization and decreased phosphorylation of both AKT and ERK1/2 in the presence of neuregulin-1 (NRG-1). In contrast, CHO cells expressing the HER3-2-2 chimeric receptor resulted in a total loss of downstream AKT activation in response to NRG-1, but maintained partial activation of ERK1/2. The results demonstrate that the intracellular domains play a crucial role in HER3's function as an allosteric activator and its role in downstream signaling.
Amino Acid Sequence ; Animals ; CHO Cells ; Cell Movement ; Cell Proliferation ; Cricetinae ; Cricetulus ; Extracellular Signal-Regulated MAP Kinases ; metabolism ; Humans ; Intracellular Space ; enzymology ; MAP Kinase Signaling System ; Models, Molecular ; Molecular Sequence Data ; Phosphatidylinositol 3-Kinases ; metabolism ; Protein Multimerization ; Protein Structure, Quaternary ; Protein Structure, Tertiary ; Proto-Oncogene Proteins c-akt ; metabolism ; Receptor, ErbB-2 ; chemistry ; Receptor, ErbB-3 ; chemistry ; genetics ; metabolism ; Recombinant Fusion Proteins ; chemistry ; genetics ; metabolism ; Signal Transduction

Objective: To study the incidence and clinicopathological features of intermediate fibroblastic/myofibroblastic tumors(IF/MFT) in infants and the young children. Methods: All available cases with soft tissue tumors in infants and children were retrieved from the files of Women and Children′s Hospital Affiliated to Qingdao University, from January 2012 to December 2017.The incidence rate of IF/MFT was observed.Cases of IF/MFT were identified and investigated by light microscopy and immunohistochemistry by reviewing the related literature. Results: Among 290 soft tissue tumors, 15 cases(5.2%) were IF/MFT, accounted for 88.2%(15/17 cases) of borderline soft tissue tumors.Twelve cases were male, 3 cases were female, the median age was 8 months, and 4 cases were congenital.Clinically, 11 cases were presented with slow-growing painless masses located in the trunk or extremities.According to histopathology, 9 cases(60.0%) were categorized as infantile fibromatosis(IFM), including 5 cases(33.3%) desmoid-type and 4 cases(26.7%) diffuse-type; 3 cases(20.0%) as lipofibromatosis(LFM); 2 cases(13.3%) as infantile fibrosarcoma(IFS) and 1 case(6.7%) as giant cell fibroblastoma(GCF). All 15 tumors were characterized by the presence of spindle fibroblasts and myofibroblasts with infiltration of the surrounding structures.Immunohistochemically, all the 15 cases were diffusely positive for Vimentin(Vim), but negative for Myogenin, MyoD1, Desmin and S-100.Smooth muscle actin(SMA), β-catenin and Bcl-2 were positive in some cases to a certain degree.The Ki-67 proliferation index was higher in diffuse-type IFM and IFS, the former was 5.0%-20.0%, and the latter was about 20.0%, however, the other cases showed Ki-67 <5.0%.The main clinical treatment was complete or extensive excision. Conclusions IF/MFT accounts for a high proportion of intermediate soft tissue tumors in infants and young children, mostly seen in male children, and IFM and LFM are the main types.The clinical signs and symptoms associated with these tumors are often nonspecific, and their histopathologic manifestations may overlap.The final diagnosis of IF/MFT must depend on the characteristics of age, location, histopathologic changes and immunohistoche-mical findings.

Objective: To establish a simple, reliable and reproducible animal model of neonatal hypoxic-ischemic encephalopathy (HIE) with similar clinical pathological process to neonates. Methods: Seven days after birth, 180 Sprague-Dawley (SD) rats were randomly divided into six groups: blank control group, experimental control group and four hypoxia groups (8, 10, 12 and 14 min hypoxia groups). Those in the experimental groups were locally anesthetized with 5% lidocaine to separate their tracheas through blunt dissection, followed by tracheal clamping with vascular clamp for 8, 10, 12 and 14 min, respectively. Rats in the experimental control group were only treated with blunt dissection of trachea. No intervention was given to the blank control group. Due to significant reduction in rat survival rate after 14 min of hypoxia, no further morphological or behavioral examination was performed in this group. Rat brain tissue sections were stained with hematoxylin-eosin (HE) 12 h after modeling. Three days after modeling, the rat brain was weighted and the apoptosis of neural cells was detected with terminal deoxynucleotidyl transferase(TdT) mediated dUTP nick end labeling (TUNEL). Morris water maze was used to screen cognitive impairment in these rats at the age of two months. One-way analysis of variance was used for statistical analysis. SNK test and Dunnett 's T3 test were performed to compare homogeneous and non-homogeneous data between groups. Results: Systemic cyanosis, loss of consciousness, paled body, urinary and fecal incontinence, twitching of the limbs and tail and other abnormal behavior were induced by hypoxia. Ischemic necrosis, bleeding, nucleus shrinkage in a large number of neurons and hyperchromatic nuclei were observed in the 8, 10 and 12 min hypoxia groups. Three days after modeling, brain weights of rats in the 8, 10 and 12 min hypoxia groups were lower than those of the blank control group and experimental control group [(1.16±0.07), (1.04±0.06), (0.97±0.12), (1.31±0.06) and (1.28±0.09) g, F=36.437, P<0.001]. However, the numbers of apoptotic cortical [(22.83±4.52), (30.25±3.02), (39.18±5.04), (7.96±2.24) and (8.86±2.49)/400 scope field, F=164.532, P<0.001] and hippocampal CA3 neurons of that three hypoxia groups were higher than those of the two control groups [(14.63±2.26), (20.25±3.02), (24.81±1.98), (4.75±2.66) and (6.67±1.78)/400 scope field, F=141.026, P<0.001]. At the age of two months, rats in the 8, 10 and 12 min hypoxia groups had longer escape latency [(17.99±6.48), (23.07±9.90), (38.94±32.46), (14.37±6.06) and (12.78±7.21) s, F=26.912, P<0.001] and fewer times of platform crossings than those in the control group and experimental control group [(5.00±1.41), (4.90±1.29), (3.75±1.83), (7.57±1.16) and (7.14±1.15) times, F=14.336, P<0.001]. Conclusions Pathological changes in brain tissues and behaviors of rats after modeling are in line with the characteristics of classic animal model of HIE and similar to the clinical pathology and physiology of HIE, and this could be a new, simple, reliable and reproducible animal model of HIE, being capable of controlling the duration of hypoxia accurately.

Objective: To verify the safety and efficacy of IONTRIS particle therapy system (IONTRIS) in clinical implementation. Methods: Between 6.2014 and 8.2014, a total of 35 patients were enrolled into this trial: 31 males and 4 females with a median age of 69 yrs (range 39-80). Ten patients had locally recurrent head and neck tumors after surgery, 4 cases with thoracic malignancies, 1 case with hepatocellular carcinoma, 1 case with retroperitoneal sarcoma, and 19 cases with non-metastatic prostate carcinomas. Phantom dose verification was mandatory for each field before the start of radiation. Results: Twenty-two patients received carbon ion and 13 had proton irradiation. With a median follow-up time of 1 year, all patients were alive. Among the 16 patients with head and neck, thoracic, and abdominal/pelvic tumors, 2, 1, 12, and 1 cases developed complete response, partial response, stable disease, or disease progression, respectively. Progression-free survival rate was 93.8% (15/16). Among the 19 patients with prostate cancer, biological-recurrence free survival was 100%. Particle therapy was well tolerated in all 35 patients. Twenty-five patients (71.4%) experienced 33 grade 1 acute adverse effects, which subsided at 1 year follow-up. Six (17.1%) patients developed grade 1 late adverse effects. No significant change in ECOG or body weight was observed. Conclusions IONTRIS is safe and effective for clinical use. However, long term follow-up is needed to observe the late toxicity and long term result.