Objective To investigate the effect of brain microvascular endothelial cells (BMECs) on mesenchymal stem cells (MSCs) expressing fetal liver kinase-1(Flk-1) and von Willebrand factor (vWF) under normal and hypoxic conditions. Methods Human bone marrow MSCs and rat BMECs were isolated, cultured and identified. We used direct and indirect co-culture to induce the differentiation of MSCs. The expression of Flk-1 and vWF after induction was observed and analyzed by flow cytometry and immunofluorescence staining. Results Undifferentiated MSCs did not express Flk-1 or vWF. In indirect co-culture group, indirect co-culture under hypoxic conditions induced (7.58?0.58)% (n=6, P=0.034) MSCs to express Flk-1 but not vWF, while in normoxic indirect co-culture, MSCs expressed neither. In direct co-culture group, after 5-day direct co-culture with BMECs under normoxia or hypoxia, MSCs expressing Flk-1 accounted for (13.76?1.67)% (n=6, P

AIM:To investigate the paracrine actions of mesenchymal stem cells(MSCs)to brain microvascular endothelial cells(BMECs)in hypoxic conditions.METHODS:Human bone marrow MSCs and rat BMECs were isolated,cultured and identified.The contents of VEGF and MMP-9 were confirmed using ELISA assays of the conditioned media.The transendothelial electrical resistance(TEER)was detected to reflect the permeability of the BMECs monolayer.The effects of different conditioned media on BMECs were investigated in hypoxic conditions.RESULTS:The contents of VEGF and MMP-9 in conditioned media were induced to increase significantly by the hypoxic stress,which were much lower in BMECs conditioned media than that in MSCs conditioned media.Hypoxia induced a significant increase in VEGF and MMP-9 in MSCs conditioned media collected in comparison to those in normal conditions.In hypoxic conditions,MSCs conditioned media enhanced the proliferation(optical density values:0.947?0.103 vs 0.254?0.024,P

AIM To assess antidiabetic and insulin sensitizing effect of a novel thiazolidinedione,neoglitazone, in different animal models of type 2 diabetes. METHODS Neoglitazone combined with low-dose insulin(0.2U·d-1 per mouse,sc)were given in streptozotiocin (STZ)-induced diabetic mice for 7 d to inspect its insulin-sensitive improvement. The antidiabetic effect of chronic oral treatment (8 wk) with neoglitazone on spontaneous diabetes mice (DB/DB mice) was examined. The levels of blood glucose were measured by a One-Touch blood glucose meter and pathology of heart, kidney, and pancreas tissues were observed under a light microscope. The hyperinsulinaemic-euglycaemic clamp technique was applied to measure the increase of glucose infusion rate (GIR) of neoglitazone on the immune insulin-resistant rats induced by Bacillus Calmette-Guerin therapy decreased ( 14±s4)%, (55±24)%,and (28±7)% of blood glucose levels compared with CMC-Na+insulin group, respectively (P < 0.01) . In DB/DB mice, neoglitazone showed better reduction in blood glucose levels than those of model animals (P < 0.01) , and pathological studies indicated that neoglitazone attenuated the diabetic kidney and pancreas lesions. During a hyperinsulinaemic-euglycaemic clamp,neoglitaxone(10,30,and 100mg·kg-1·d-1,ig for 2 wk )-treated groups required significantly higher GIR to maintain basal glucose concentrations than model group (P < 0.01 ). CONCLUSION Neoglitazone could directly enhance insulin sensitivity and ameliorate insulin resistance in different diabetic animal models.

Objective:To explore the mRNA differential expression and clinical effect of leptin in different kinds of ovarian cancer.Methods: 129 patients with ovarian cancer and 42 controls were collected from Jun.2015 to Jun.2016 in our hospital.We got a small tissue by ovarian biopsy guided under the ultrasonic processing,detected the total RNA,purified mRNA and analyzed the correlation of the expression of leptin in different kinds and clinical stages of ovarian cancer.Results: Compared with the control groups,the mRNA expression of leptin in each group was increased(P<0.05),and the serous adenocarcinoma group increased obviously(P<0.05).Patients with clinical stage FIGO Ⅲ was significantly higher than other stages in serous adenocarcinoma group(P<0.05).Conclusion: Leptin could express in different kinds and clinical stages of ovarian cancer,and may be a new detection index for early diagnosis of ovarian cancer.

Background and purpose: It has been reported that activation of Notch1 could strongly inhibit proliferation of HPV (human papilloma virus)-positive HeLa cells by down-regulation of the E6 and E7 genes. The aim of this paper was to investigate the role of the Notch signaling pathway in growth arrest of EC109 cells in vitro and the molecular mechanism. Methods: EC109 cell lines, a well differentiated human ESCC (esophageal squamous cell carcinoma) cell line with HPV18-positive, was used in the study. Exogenous intracellular domain of Notch1(ICN) was transfected into cultured EC109 cells by lipofectamine transfection, the proliferation of the transfected cells was measured by an MTT assay. Cell cycle distribution was analyzed by flow cytometry. Human papilloma virus type 18 (HPV18) E6/E7 mRNA expression was detected by RT-PCR, and p53 protein expression was detected by Western blot.Results: Activation of Notchl signaling resulted in inhibition of EC109 cell proliferation with the induction of G_2/ M arrest. There was a significant difference in terms of the percentage of G_2/M phase cells among the ICN-transfected group (42.57±1.57)% and the non-transfected group (1.88±0.66)% or the empty plasmid transfected group (1.99±1.02)% (P<0.01). Down modulation of HPV18 E6/E7 gene expression and upregulation of p53 expression was (2.15±0.23) in ICN-transfected group higher than non- transfected group (0.45±0.07) and empty plasmid transfected group (0.46±0.02) (P<0.01). Conclusion: Repression of HPV18 E6/E7 expression by Notch1 signaling results in growth suppression of HPV18-positive EC109 cells with concomitant activation of p53-mediated pathways.

ObjectiveTo investigate the clinical,myopathological and molecular changes in two Chinese families with oculopharyngodistal myopathy ( OPDM).MethodsWe performed muscle biopsy and histopathologic study on the probands of two families,and further examined molecular genetic testing on PABPN1 and GNE gene. Results Family 1 included 3 affected brothers in the same generation and family 2 involved 4 patients in 2 generations. Dysarthria rather than external ophthalmoplegia was the prominent oculopharyngeal symptoms for Chinese patients. No intranuclear inclusions were observed in ultrastructural examination.The number of GCG repeats in the PABPN1 gene was within normal range and no mutations were identified in the GNE gene.ConclusionsFamily 1 is the first publication on autosomal recessive OPDM in China.The age of onset of two families was comparable with Japanese patients and the pattern of muscle involvement was different. OPDM is a distinct phenotypical,histological,and genetic entity.

Objective:To evaluate the differences of clinical parameters and putative periodontal patho-gens in sites of different probing depth ( PD) reduction after non-surgical periodontal treatment in patients with aggressive periodontitis ( AgP ) .Methods: Clinical examinations including plaque index , probing depth (PD), attachment level (AL) and bleeding index (BI), and full-mouth periapical photographs were collected from 20 patients with AgP .All the patients received non-surgical periodontal treatment , including oral hygiene instruction , supra-gingival scaling , subgingival scaling and root planing ( SRP ) and were followed up for 6 months post-therapy.Gingival crevicular fluids (GCF) were collected at 1 site in each quadrant before and at the end of 6 months post-therapy .Six kinds of putative periodontal patho-gens and 6 kinds of short chain fatty acids ( SCFAs ) were detected in the GCF samples .Results: The baseline clinical parameters of PD , AL and BI , the baseline concentration of succinic acid , acetic acid , propionic acid and butyric acid , and the prevalence of Treponema denticola were significantly higher in sites with PD reduction more than 2 mm sites compared with PD reduction no more than 2 mm sites [(7.7 ±1.2) mm vs.(5.1 ±1.8) mm, (6.3 ±1.9) mm vs.(4.5 ±2.2) mm, 3.8 ±0.4 vs.3.3 ± 0.8, 1.66 mmol/L vs.1.10 mmol/L, 31.67 mmol/L vs.17.78 mmol/L, 3.31 mmol/L vs.1.95 mmol/L, 84.6%vs.56.1%, P<0.05].However, there were no significant differences in the clinical param-eters, the 6 kinds of putative periodontal pathogen detection and SCFAs concentration between the 2 groups at the end of 6 months post-treatment.In sites with PD>5 mm at the end of 6 months post-thera-py , all were found with red complex bacteria infection .Conclusion:The baseline clinical parameters are important factors in predicting PD reduction after non-surgical periodontal treatment in patients with AgP . In sites with deep pockets after non-surgical periodontal treatment , the active control of red complex bac-teria is recommended .

Objective This study explored the expression of polyclonal protein SUZ12 in patients with intrahepatic cholangiocarcinoma(ICC),and its role in predicting the survival and treatment of ICC patients.Methods The expression of SUZ12 and p16INK4a was detected by immunohistochemical assay in 207 liver tissue samples including ICC patients,BilIN-1,-2,-3 and non-tumor-like cholangiocarcinoma.The expression of these proteins was assessed to be related to the pathological characteristics of the ICC patients receiving chemotherapy and the outcome of survival as well as the subsequent chemotherapy response.Results The expression level of SUZ12 was gradually increased from non-neoplastic bile duct tissue to BilIN-1,-2,-3 and ICC.The expression of p16INK4a protein was expressed in non-neoplastic-like cholangiocarcinoma,but it decreased gradually in BilIN-1,-2,-3 and ICC tissues.SUZ12 expression was associated with undifferentiated ICC,lymph node metastasis and advanced cancer.Kaplan-Meier curve analysis showed that ICC patients with high expression of SUZ12 had a significant reduction in overall survival and disease-free survival in comparison with ICC patients with the low expression of SUZ12.SUZ12 expression was significantly associated with overall survival of patients receiving adjuvant gemcitabine-based chemotherapy(AGC).Conclusion SUZ12 expression is able to predict the overall survival and disease-free survival of ICC patients with adjuvant gemcitabine-based chemotherapy.

Objective:To analyze the serum IgG titers to Aggregatibacter actinomycetemcomitans ( Aa ) and associated factors in patients with aggressive periodontitis (AgP).Methods:Venous blood samples were collected from 62 AgP patients and 45 periodontal healthy controls , unstimulated whole saliva and pooled subgingival plaque samples of AgP patients were also collected for the detection of Aa ( PCR method) .Serum IgG titers to Aa serotype c were measured by enzyme-linked immunosorbnent assay ( ELISA) .Results:The detection rates of serum IgG to Aa serotype c in the AgP patients and the healthy controls were both 100%.The AgP patients exhibited significantly higher IgG titers to Aa serotype c than the healthy controls (11.1 ±1.9 vs.9.1 ±1.8, P<0.01).There was no significant difference in serum IgG levels to Aa serotype c and in the prevalence of high-responding patients to Aa serotype c between the incisor-first molar type AgP patients and generalized AgP patients .Serum IgG titers to Aa serotype c in the Aa-positive AgP patients ( the patients who were Aa-positive in subgingival plaque or saliva ) were sig-nificantly higher than those of the Aa-negative patients (11.9 ±1.3 vs.10.7 ±2.1, P<0.05).Con-clusion:Serotype c was the main serotype of Aa in Chinese patients with AgP .Serum IgG responses in generalized AgP patients were comparable to those in incisor-first molar type AgP patients .

Objective To observe the seroreversion of serological tests for syphilis in neonates born to mothers with syphilis and to evaluate the influence of antisyphilitic treatment on the time of seroreversion in uninfected neonates.Methods A total of 115 neonates born to mothers with syphilis were recruited in this study,and underwent toluidine red unheated serum test (TRUST),Treponema pallidum particle assay (TPPA)and 19S IgM-TPPA at birth.Follow up was scheduled at 1,3,6,9,12,18 and 24 months of age,and cancelled as soon as both TRUST and TPPA became negative during the 24 months.Both TRUST and TPPA were carried out at each follow-up visit.Kaplan-Meiter survival analysis and Wilcoxon rank-sum test were conducted to assess these data by using SPSS 13.0 software.Results At birth,TPPA was positive in all (100％) of the neonates,TRUST in 97 neonates (84.3％) with the titer ranging from 1 ∶ 1 to 1 ∶ 256,19S IgM-TPPA in 3 neonates who were diagnosed with congenital syphilis (CS).CS was ruled out in the other 112 neonates with negative 19S IgM-TPPA after follow up.Of the 3 neonates with CS,all became negative for TRUST within 12-18 months after birth,2 remained positive for TPPA at 24 months of age,1 remained positive for TPPA at 20 months of age,and all were still followed at the time of this writing.The cumulative seroreversion rate was 82.2％,98.9％ and 100％for TRUST at 0-3,4-6 and 7-9 months of age,respectively in 90 uninfected neonates,3.1％,30.2％,85.4％ and 100％ for TPPA at 0-3,4-6,7-9 and 10-12 months of age,respectively in 96 uninfected neonates.Of 16 neonates who missed some follow-up visits,all turned negative for TRUST and TPPA within 15 months after birth.As far as the TP-seropositive uninfected neonates were concerned,no statistical differences were observed in the seroreversion time of TRUST or TPPA between neonates receiving antisyphilitic treatment and those without antisyphilitic treatment (x2 =0.54,2.41,respectively,both P ＞ 0.05).The seroreversion of TRUST occurred earlier than that of TPPA (Z =10.45,P ＜ 0.01).Conclusions Most uninfected neonates born to syphilitic mothers turn negative for TRUST within 6 months after birth,and for TPPA within 12 months.And the seroreversion of TRUST usually occurs earlier than that of TPPA.Antisyphilitic treatment shows no significant influence on the seroreversion time of TRUST or TPPA in these seropositive uninfected infants.