To explore the feasibility of mesenchymal stem cells (MSCs) acting as seed cells in tissue engineering, we isolated human bone marrow MSCs and differentiated them into vascular endothelial-like cells (ELCs) in vitro. Bone marrow mononuclear cells (BMSCs) were isolated by the method of percoll density centrifugation, and seeded in Dulbecco Modified Eagle Medium supplemented with 10% fetal bovine serum. MSCs were purified through multiple adherent cultures, and differentiated into ELCs induced by endothelial cell growth medium-2 (EBM-2) medium containing vascular endothelial growth factor (VEGF), human fibroblast growth factor (hFGF), insulin like growth factors 1 (IGF-1), and human epidermal growth factor (hEGF). The relative biologic characteristics of ELCs including cell morphology and phenotype were studied by inverted microscope and flow cytometry. The induced cells were identified by immunofluorescence with CD31 and Von Willebrand factor (vWF). The results showed that the morphology of MSCs was long-spindle and vortex-like growth. After induction of differentiation, the cells were round, and similar to vascular endothelial cells (ECs). Flow cytometric analysis revealed that ELCs expressed ECs specific surface markers of CD31 and vascular endothelial cadherin (VE-cadherin), but not CD133. Immunofluorescence results also confirmed that ELCs expressed CD31 and vWF. The results suggested that ELCs possed similar cell biological characteristics with ECs. In one word, human MSCs derived from bone marrow have the potential to differentiate into ECs in vitro, and show clinical feasibility acting as ideal donor cells of vascular tissue engineering.
Antigens, CD ; metabolism ; Bone Marrow Cells ; Cadherins ; metabolism ; Cell Culture Techniques ; Cell Differentiation ; Cells, Cultured ; Culture Media ; chemistry ; Endothelial Cells ; cytology ; Epidermal Growth Factor ; pharmacology ; Fibroblast Growth Factors ; pharmacology ; Flow Cytometry ; Humans ; Insulin-Like Growth Factor I ; pharmacology ; Mesenchymal Stromal Cells ; cytology ; Platelet Endothelial Cell Adhesion Molecule-1 ; metabolism ; Tissue Engineering ; Vascular Endothelial Growth Factor A ; pharmacology ; von Willebrand Factor ; metabolism

Objective To investigate the relationship between the quality of life and different frequency of sudden death in Yunnan unexplained sudden death disease area.Methods According to the stratified cluster sampling method,728 individuals were selected as the respondents in Heqing County,Eryuan County of Dali Bai Autonomous Prefecture,and in Dayao County of Chuxiong Yi Autonomous Prefecture.According to the random sampling,649 individuals were selected as the control in Yongping County,Dali Bai Autonomous Prefecture.Data of the quality of life (WHOQOL-BREF version) were collected through household surveys.Analysis method including ANOVA,Chi-squared test and multilinear regression were used.Results Compared with the control population,the household income of population in the diseased area was not significantly different statistically (x2 =7.052,P ＞ 0.05).But the differences in education level and chronic disease situation were statistically significant (x2 =35.727,9.810,all P ＜ 0.05).According to the frequency of the sudden death,from one to four,the total scores of the quality of life,the scores of the physiology domain,the scores of the psychological domain,the scores of the environmental domain and the scores of the social relations domain (1 time:54.30,13.74,14.43,11.21,14.91;2 times:54.75,13.86,14.65,11.12,15.10;3 times:52.40,13.21,13.76,11.00,14.41;4 times:49.21,12.15,12.54,9.87,14.64)were all lower than those of the control group (56.03,14.11,14.78,11.88 and 15.26),the differences between two groups were statistically significant (x2 =41.88,25.75,41.07,35.07,8.08,all P ＜ 0.05);the total scores and each domain score of the quality of life were negatively correlated with the frequency of sudden death (the multi-variables regression coefficient were as follows:-1.195,-0.341,-0.356,-0.314,and-0.185,all P ＜ 0.05).Conclusions The quality of life of those who have lived in Yunnan unexplained sudden death area is associated with the outbreak frequency of sudden death.Following increasing of the outbreak frequency of Yunnan unexplained sudden death,the quality of life of the population living in Yunnan unexplained sudden death area has decreased.

Objective To evaluate the relieving effects of vanillin sinffing on depression-like behaviors in depressed rats and to explore the possible underlying mechanism.Methods Depression animal model established by chronic unpredictable medium intensity stress combined with isolation and destroy the olfactory bulb.The depressed rats were divided randomly into vanillin inhalation group,fluoxetine hydrochloride group,depression model group,olfactory bulbectomy with the vanillin inhalation treatment group and sham-operated group.Nervous behavioral changes had been observed at different time after the administration of 5 weeks.The concentration of brain derived neurotrophic factor(BDNF) in the brain homogenate and the positive expression of BDNF in hippocampus had been also measured.Results Two weeks after the intervention,the immobility time of vanillin group((12.78 ±7.50) s) was lower than that of the model group((57.33±32.16) s) (P＜0.05).The consumption of saccharose in vanillin group((52.88±25.18)g) was higher than that of model group((37.40±19.33) g) (P＜0.05).BDNF of the brain homogenate in vanillin group (0.54±0.13) was significantly increased compared with model group (0.36± 0.06) (P＜0.01).When compared with the OBX group (0.40±0.06),similar result was obtained.Immunohistochemistry and the average density of image analysis revealed that the expression of BNDF of hippocampal CA3 in vanillin group (0.40±0.03)was significantly increased compared with model group (0.25±0.04) and OBX group (0.28±0.03) (P＜0.01).Conclusion Vanillin inhalation significantly relieves depression-like behaviors in depression rats.The possible mechanism may increase hippocampal neurogenesis by raising brain derived neurotrophic factor in brain.

Due to the good tumor-targeting and excellent biocompatibility, the drug-loading nanoparticles (NPs) has been widely applied in the diagnosis and treatment of cancer. However, after the NPs are recognized and internalized by cancer cells, the effects of NPs on cell migration behavior were unclear. In the present study, the self-assembly techniques (SAMs) was used to modify gold (Au) nanoparticles (Au NPs) with different chemical functional groups (CH3, OH, COOH and NH2) as model NPs. The dispersion of these groups in solution and the distribution in cells were studied by transmission electron microscope (TEM), respectively, and the proliferation was examined by MTT assay in vitro. The wound-healing and the Transwell assay were used to examine the effect of internalized Au-NPs on HepG2 cells migration. The results showed that different Au-NPs mainly distributed at the edge of the vesicle membrane and the gap between cells. The Au-NPs resulted in decreased cell viability in a concentration-depended manner. In addition, the results of wound-healing and Transwells assay indicated that the internalization of the NH2-NPs and OH-NPs would inhibit cell migration compared with those in the control group.
Carcinoma, Hepatocellular ; metabolism ; Cell Movement ; Cell Proliferation ; Cell Survival ; Gold ; Hep G2 Cells ; Humans ; Liver Neoplasms ; metabolism ; Metal Nanoparticles ; chemistry

This paper is aimed to investigate the effect of fluid shear stress on the tight junction of laryngeal squamous carcinoma (Hep2) cells and to explore the potential molecular mechanism. Hep2 cells were selected and subjected to the fluid shear stress of 1.4 dyn/cm2 for different time, respectively. The morphological changes of Hep2 cells under shear stress were observed using inverted microscope. The cell-cell junctions were examined by transmission electron microscope (TEM). The expressions of tight junction proteins (including Occludin, Claudin-5 and ZO-1) and the distribution of Claudin-5 were examined by Western blot assay and laser scanning confocal microscope, respectively. The results indicated that Hep2 cells turned to spindle-like shapes after exposed to shear stress, and showed the trend of the recovering to original shapes when the shear stress was cancelled. The cell-cell junctions were tight under the shear flow condition, and the permeability was reduced under the condition of 1.4 dyn/cm shear flow. The expressions of tight junction proteins were enhanced with increased duration of shear flow, but reduced after removing shear flow. The result of Claudin-5 expression by immufluorescence assay was consistent with that by Western blot. The Claudin-5 mainly distributed in the cytoplasm under static condition, while it located at the intercellular after shear flow stimulation, and it appeared intercellular and cytoplasm after stopping shear flow stimulation. Therefore, it can be concluded that shear stress changes the morphology of laryngeal squamous carcinoma Hep2 cells, and upregulates the tight junction.
Blotting, Western ; Carcinoma, Squamous Cell ; pathology ; Claudin-5 ; metabolism ; Hep G2 Cells ; Humans ; Laryngeal Neoplasms ; pathology ; Occludin ; metabolism ; Stress, Mechanical ; Tight Junctions ; Zonula Occludens-1 Protein ; metabolism

Objective To explore the value of distinguishment of hepatocellular carcinoma (HCC), cirrhosis nodules and normal liver based on neural networks in the ~(31)P-MR spectroscopy. MethodsA total of 66 data of ~(31)P-MRS were analysed using back-propagation neural network, including 37 samples of liver cirrhosis, 13 samples of HCC and 16 samples of normal liver. ResultsThe cross-valiation experiments showed that diagnostic accuracy rate of HCC increased from 85.47% to 92.31% with neural network model based on the ~(31)P-MR spectroscopy data analysis. Conclusion ~(31) P-MRS data analysis based on neural network model provides a valuable diagnostic tool of HCC in vivo.

To study the potential molecular mechanism of tumor angiogenesis in its microenvironment, we investigated the effects of HepG2 conditioned medium on the proliferation of vascular endothelial cell and vascular angiogenesis in our laboratory. Human umbilical vein endothelial EA. hy926 cells were co-cultured with HepG2 conditioned medium in vitro. The proliferation and the tubulogenesis of EA. hy926 cells were detected by teramethylazo salt azole (MTT) and tube formation assay, respectively. The results showed that the survival rate of the EA. hy926 cells was significantly increased under the co-culture condition. HepG2 conditioned medium also enhanced the angiogenesis ability of EA. hy926 cells. In addition, the expressions of intracellular VEGF and extracellular VEGFR (Flk-1) were regulated upward in a time-dependent manner. In conclusion, the proliferation of vascular endothelial cells and Vascula angiogenesis were improved under the condition of indirect co-culture.
Carcinoma, Hepatocellular ; pathology ; Cell Proliferation ; Coculture Techniques ; Culture Media, Conditioned ; Endothelial Cells ; cytology ; Hep G2 Cells ; Human Umbilical Vein Endothelial Cells ; Humans ; Liver Neoplasms ; pathology ; Neovascularization, Pathologic ; Tumor Microenvironment ; Vascular Endothelial Growth Factor A ; metabolism ; Vascular Endothelial Growth Factor Receptor-2 ; metabolism

BACKGROUND:At present, there is no effective treatment strategy for cavernous transformation of portal vein and basic research about its etiology is rarely reported.
OBJECTIVE:To establish the models of cavernous transformation of portal vein, detect the expression of matrix metal oproteinase-2,-9 (MMP-2, MMP-9) and tissue inhibitors 1, 2 of metal oproteinase (TIMP-1, TIMP-2) in rat portal vein and peripheral tissue, and discuss the roles in the process of peripheral angiogenesis.
METHODS:Eighty Sprague-Dawley rats were randomly divided into three groups. The rat models of cavernous transformation of portal vein were established with partial coarctation in portal vein by using 21 G blunt pinhead. Control group was normal rats without operation (samples were harvested after portal vein radiography). Model group and sham operation group were divided into three groups respectively according to different time points, namely 2, 4 and 6 weeks after operation. Rats of each group were randomly chosen at week 2, 4 and 6 after operation to observe the formation of col ateral circulation of portal vein and its peripheral tissues by performing portal vein radiography. CD31 was detected by immunohistochemistry. The expression of MMP-2, MMP-9, TIMP-1, TIMP-2 mRNA and protein in portal vein and peripheral tissue were determined by RT-PCR and immunohistochemistry respectively.
RESULTS AND CONCLUSION:Peripheral angiogenesis of model group was increased obviously by portal vein radiography and immunohistochemistry. RT-PCR and immunohistochemistry results demonstrated that, compared with the control group and sham operation group, the expression of MMP-2 mRNA and protein in model group were significantly increased at weeks 2, 4, and 6 (P<0.01, P<0.05), while expression of MMP-9 mRNA and protein at week 2 in model group were significantly higher than that in the control group and sham operation group. Expression of TIMP-1 and TIMP-2 in model group showed no significant difference compared with control group and sham operation group at weeks 2, 4, and 6 (P>0.05). Ratio of MMP-2/TIMP-2 of model group was significantly higher than that of control group and sham operation group (P<0.05) at week 2. the rat models of cavernous transformation of portal vein have low mortality, high success rate and are stable. Upregulation of the expression of MMP-2, MMP-9 and the disbanlance of the ratio of MMP-2/TIMP-2 might contribute to the peripheral angiogenesis in rats with cavernous transformation of portal vein.

Objective To compare the effects of two skin sampling methods (negative pressure suction blister and skin shaving) on the physical status of autologous epidermal keratinocytes transplanted to patients with vitiligo. Methods Skin samples were obtained from the normal skin of 32 patients with stable vitiligo by suction blister under negative pressure and skin shaving alone or in combination. Immunohistochemistry was performed to examine the expression of proliferating cell nuclear antigen (PCNA) and caspase-3 in these samples.Skin tissues resected from 15 normal human subjects served as the control. Results There was an expression of PCNA and caspase-3 at different degrees in all the skin tissues obtained by the two sampling methods from the 32 patients. Most PCNA-positive cells were focally distributed at the basal layer in epidermis obtained from suction blisters, and a few PCNA-positive cells were observed in the middle and lower part of the prickle cell layer of epidermis from shaved skin. There was a significant difference in the percentage of PCNA-positive cells between the epidermis from suction blisters and shaved skin as well as between the epidermis from suction blisters and normal control skin (x2 = 10.99, 14.08, both P < 0.05), but not between the epidermis from shaved skin and normal control skin (x2 = 1.31, P > 0.05). The expression of caspase-3 was predominantly located in the cytoplasm of keratinocytes in the basal layer as well as middle and lower part of prick cell layer, and no difference was observed in the percentage of caspase-3-expressing keratinocytes between the epidermis from shaved skin, suction blisters and normal control skin (x2 = 1.41, 2.89, 1.91, all P > 0.05). Conclusions The proliferation activity of epidermal cells seems important to the survival of grafted skin, and compared to the suction blister technique, skin shaving appears to have less influence on the proliferation of keratinocytes.

Six atrazine-degrading strains, Pseudomonas spp. AD1, AD2, AD6, Agrobacterium sp. AD4, Xanthomonas AD5, and Erwinia sp. AD7, were isolated from industrial wastewater. These strains are able to grow on atrazine as sole nitrogen source. Strain AD1 is able to degrade atrazine of 0. 3g/L in minimal medium at a percentage of 99.9% in 72 hours. PCR products that are homologous to the atrazine chlorohydrolase gene atzA)from Pseudomonas sp. strain ADP were obtained by PCR method using total DNA of the strains AD1 ,AD4,AD5,AD6,and AD7 as templates.