Objective: To study the effects of endostatin and doxycycline on microcirculatory patterns in the melanoma transplant model of the chorioallantoic membrane of the chicken embryo and the experimental evidences for melanoma therapy thereof. Methods: Endostatin was dripped on the chorioallantoic membrane after melanoma was transplanted 3 days (A group) and 5 days (B group). Doxycycline was dripped on the chorioallantoic membrane after melanoma was transplanted 3 days (C group) and 5 days (D group). The PBS solution was dripped on the chorioallantoic membrane in control group (E group). The hematoxylin-eosin（HE）staining and immunohistochemical staining were performed to observe microcirculation patterns in sections. Results: Compared with the E group, the area of endothelium-dependent vessels were significantly decreased in A group and B group (P ＜ 0.01). But the areas of vasculogenic mimicry were bigger in the two groups(P ＜ 0.05). Compared with E group, the areas of endothelium-dependent vessels and vasculogenic mimicry were significantly decreased in D group(P ＜ 0.05). Conclusion: Endostatin can inhibit angiogenesis. But it has no effect on vasculogenic mimicry in the transplant melanoma. And Doxycycline can inhibit vasculogenic mimicry and angiogenesis in the transplant melanoma. The results provided experimental evidences for melanoma therapy.

Objective:To investigate the effect of DKK1 on linearly patterned programmed cell necrosis (LPPCN) and vasculogenic mim-icry (VM) and the related molecular mechanism in non-small cell lung cancer (NSCLC). Methods:A total of 173 human NSCLC speci-mens were collected to detect LPPCN by H&E staining, detect VM with CD31/PAS double staining, and investigate DKK1 and related protein expression by immunohistochemistry. The clinical pathological significance of LPPCN, VM, and DKK1 and the correlation of them were analyzed. Human NSCLC H460-DKK1 cells were engrafed in nude mice to evaluate the influence of DKK1 up-regulation on VM and LPPCN in vivo. Results:Approximately, 14.45%(25/173) of NSCLC had VM and 49.71%(86/173) had LPPCN. 25.6%(22/86) of NSCLC cases in LPPCN-positive group formed VM. Both of VM and LPPCN were all correlated with poor differentiation, late TNM stage, easy recurrence and metastasis and poor prognosis in NSCLC. DKK1 expression in the VM-positive group and the LPPCN-positive group was higher than that in the VM-negative group and the LPPCN-negative group, respectively. DKK1, LPPCN, and VM were positive-ly correlated with VE-cadherin, MMP-2,β-catenin nuclear expression and Twist1. H460-DKK1 transplantation tumor model confirmed that DKK1 promotes the expression of VM and LPPCN and related proteins in NSCLC. Conclusion:The increase of theβ-catenin and Twist1 expression induced by DKK1 may promote the formation of LPPCN and VM in NSCLC.

Objective:To discuss the effects of recombinant IFN-α-IL-18 fusion protein on chicken lymphocyte histamine induced and the NF-κB p65 activation and nucleo-cytoplasmic transport.Methods: The healthy chickens blood was sterile adopted with anticoagulant,then separation of the chicken peripheral blood lymphocyte and divided into 10 groups:The yeast expression and purification protein IFN-α-IL-18,IL-18,IFN-α were added with 250 ng/ml,500 ng/ml and 1 000 ng/ml respectively while the control was only added RPMI1640 with 3 repetitions for each group.Then the histidine decarboxylase activity,histamine,IFN-γ,PI3K,MAPK and NF-κB p65 in cell nucleus were detected.Results: The recombinant IFN-α-IL-18 and IL-18 could significantly promote the activity of histidine decarboxylase (P<0.01),increase the contents of histamine (P<0.01),induce IFN-γ (P<0.01),improve the contents of PI3K (P<0.01) and the NF-κB p65 levels in nucleus (P<0.01),and the higher concentration of IFN-α had a similar effect to lymphocytes.The effects of IFN-α-IL-18,IL-18 and IFN-α on MAPK was acratia.Conclusion: IFN-α-IL-18 and IL-18 can stimulate chicken peripheral blood lymphocyte populations increased histamine contents significantly and promote the induction of IFN-γ.IFN-α-IL-18,IL-18 and IFN-α increase PI3K expression in lymphocyte associated with the NF-κB activation and NF-κB p65 nucleo-cytoplasmic transport.The study built foundation for the function of IFN-α-IL-18 and the exploration of the mechanism of controlling epidemic diseases.

Objective To investigate the cerebral haemodynamics change and morbidity of periventricular-intraventricular hemorrhages(PIVH) in premature infants with or without patent ductus ateriosus (PDA).Methods Bedside Echocardiography and transcranial sonography (TCS) were performed on 85 cases of preterm infants in 48 h,48 to 96 h,96 to 120 h,120 to 168 h after birth (gestational age≤34 weeks and birth weight≤2 000 g).PDA,peak systolic velocity (Vs),end-diastolic velocity (Vd),pulsatility index (PI) and resistant index (RI) of anterior and middle cerebral artery(ACA and MCA) and PIVH were monitored simutaneously.Results According to the Echocardiography and clinical symptoms,all the cases were divided into 3 groups:haemodynamic significant PDA group (hsPDA group,n =23),non-hsPDA group(nhsPDA group,n =38) and non-PDA group(nPDA group,n =24).The mean birth weight and gestational age did not differ significantly among the 3 groups.Single and multiple Logistic analysis indicated that invasive mechanical ventilation less than 48 h after birth was related to hsPDA (x2 =11.182,P =0.019; OR =10.06,P =0.039).Repetitive measurement deviation analysis found that:Vd of ACA and MCA were lower in the hsPDA group than those in the nhsPDA group (P =0.000,P =0.001) and the nPDA group (P =0.003,P =0.013),while PI and RI were higher than in the other 2 groups.Compared with non-closed group,at 7 days after birth in hsPDA group,RI and PI of closed group were significantly lower,while Vd was significantly higher.Nevertheless,there was no significant difference in hemodynamic parameters when comparison with nPDA group at 7 days after birth.There was no statistical difference in the grading and severity of PIVH with or without PDA.However,the rate of severe PIVH was found higher in hsPDA group than the other 2 groups(17.39％ vs 8.33％ vs 5.26％,x2 =2.405,P =0.280).Conclusions The invasive mechanical ventilation less than 48 h after birth was probably associated with preterm hsPDA.HsPDA could result in major cerebral haemodynamic changes and increase the morbidity of severe PIVH.

Objective:To discuss the influence of ALDH1+and CD133+phenotypic breast cancer stem-like cells in TA2 triple negative breast cancer on promoting epithelial-mesenchymal transition (EMT) occurrence in TA2 mice with triple-negative breast cancer and on their biological behavior. Methods:Flow cytometry was performed to analyze the markers ALDH1 and CD133 in TA2 mice triple nega-tive breast cancer and breast cancer stem-like cells with ALDH1+, ALDH1?, CD133+, and CD133?phenotypes, which were sorted out. Then, the TA2 mice were inoculated with sorted tumor cells according to cell type. The mice were divided into ALDH1+, ALDH1?, CD133+, and CD133-groups. The tumor-growing conditions were observed. A tumor tissue was sliced for the immunohistochemical testing of ALDH1?, CD133?, and EMT-related Twist1, E-cadherin, and VE-cadherin proteins. The expression difference of breast cancer stem cell surface markers ALDH1 and CD133 in triple-negative breast cancer and EMT-related proteins Twist1, E-cadherin, and VE-cad-herin was analyzed. Results:The expression rates of breast cancer stem cell markers ALDH1 and CD133 in TA2 mice triple negative breast cancer were 31.2%and 6.5%, respectively. The tumor growth ability of TA2 mice from ALDH1+group was obviously stronger than that from ALDH1?group. The CD133+group was evidently stronger than CD133?group. The immunohistochemical results showed that ALDH1, Twist1, and VE-cadherin expression levels in the ALDH1+group were evidently higher than that in the ALDH1?group (all P<0.05). E-cadherin expression decreased (P<0.05). CD133?, Twist1, and VE-cadherin expression levels in CD133+group were higher than that in CD133?group (all P<0.05). Conclusion:In TA2 mice triple negative breast cancer, ALDH1+and CD133+phenotypic breast cancer stem-like cells may influence the expression of EMT-related proteins, and promote the formation of triple-negative breast cancer.

The main objective of this study was to observe the adhesion, proliferation and differentiation of mouse osteblast-like MC3T3-E1 cells cultured on maleic anhydride-modified poly(D,L-lactic acid) (MPLA) and poly(D,L-lactic acid) (PDLLA) polymers, and to evaluate the cytocompatibility of MPLA polymer. The effects of MPLA and PDLLA polymers on the morphology, adhesion, proliferation, the content of total cellular protein, alkaline phosphatase (ALP) activity and the content of Ca of MC3T3-E1 cells were explored. These results indicated that MC3T3-E1 cells on MPLA polymer adhered and spread more fully. On MPLA polymer, the proliferation, total protein content, ALP activity, Ca content of the cells were significantly higher than those of the cells on PDLLA polymer (P < 0.01). It was concluded that MPLA polymer could promote the adhesion, spreading, proliferation and the synthesis of protein of osteoblasts, and also induced the differentiation and mineralization of osteoblasts, suggesting that MPLA polymer might have the better cytocompatibility than PDLLA.
Animals ; Biocompatible Materials ; chemistry ; pharmacology ; Cell Adhesion ; drug effects ; Cell Differentiation ; drug effects ; Cell Line ; Cell Proliferation ; drug effects ; Embryo, Mammalian ; Lactic Acid ; chemistry ; pharmacology ; Maleic Anhydrides ; chemistry ; pharmacology ; Mice ; Osteoblasts ; cytology ; Polyesters ; Polymers ; chemistry ; pharmacology

Objective: To determine the expression of TAZ and its role in angiogenesis in gastric carcinoma. Methods: Immunohistochemical staining was performed to investigate the expression of TAZ and to determine whether a direct relationship exists between TAZ and β-catenin. Transfection with TAZ overexpression plasmid in MKN28 cells was conducted to induce exogenous expression of TAZ and a TAZ knockdown plasmid was transfected into MGC803 cells to reduce TAZ levels. The effects on endothelial cell formation, proliferation, and migration were determined by Matrigel three-dimensional culture, MTT proliferation assay and Transwell migration assay. In addition, the expression of TAZ and β-catenin in transfected gastric cancer cells was detected by Western blot. Results: Immunohistochemistry showed that TAZ protein was expressed in 64 of 150 gastric cancer sample tissues (43%), TAZ was localized in the nucleus, and its expression was associated with tumor grade, TNM stage, metastasis, and microvessel density (MVD) (P<0.05). In addition, the expression frequency of β-catenin in the TAZ positive group was 67.2%, which was significantly higher than that in the TAZ negative group, and the expression of TAZ was positively correlated with β-catenin. After transfection, TAZ overexpression increased the expression of β-catenin and enhanced HUVECs tube formation, proliferation, and migration. In the MGC803 cells transfected with the knockdown plasmid, β-catenin levels were decreased and HUVECs motility was inhibited. Conclusions: TAZ may promote angiogenesis in gastric cancer by promoting β-catenin expression.