?AIM:To investigate the effects of Bevacizumab on the proliferation and the expression of E -Cadherin and fibronectin in human retinal pigment epithelial cell ( ARPE-19) in vitro.?METHODS: Different concentrations (0, 0.625, 1.25, 2.5, 5.0mg/mL) of bevacizumab were exposed to ARPE-19 cells, then cell viability was analyzed by CCK-8, cell cycle was determined by flow cytometry, and the expression of E-Cadherin and fibornectin was detected by Western blot and RT-PCR.?RESULTS:The concentration as 2.5mg/mL or 5.0mg/mL of bevacizumab was shown to effectively suppress the proliferation and cell cycle of ARPE-19 cell (P<0.05). In addition, 2.5mg/mL or 5.0mg/mL of bevacizumab could downregulate the expression of E-cadherin and promote the transcription of fibronection gene (P<0.05).?CONCLUSION:High concentration of bevacizumab was able to inhibit ARPE-19 proliferation, downregulate E-Cadherin expression and promote fibronectin expression, indicating epithelial-mesenchymal transition induced by bevacizumab in ARPE-19 cell.

Objective To observe the effects of CO2 pneumoperitoneum on the enteric motor function and acetylcholine esterase(AchE) neuron and nitric oxide synthase(NOS) neuron in the enteric nervous system,and explore the neuromechanism of the CO2 pneumoperitoneum on renteric motor function.Methods Thirty-six Sprague Dawley rats were randomly divided into experiment group(n=24) and control group(n=12).The experiment group was divided into two subgroups namely pneumoperitoneum 30min group and pneumoperitoneum 60min group(12 each) based on the maintenance time of pneumoperitoneum.Rats in each group were gavaged with medicinal carbon powder,and then the transmission of carbon powder in small intestine was determined.The spreading specimens of intestinal myenteric plexus of small intestine were prepared and the stained AchE and NOS neurons were observed and compared.Results The propellant velocity of carbon powder was slower in pneumoperitoneum 60min group than that in pneumoperitoneum 30min group and control group(28.55%?3.45% vs 45.90%?6.30%,48.25%?5.28%,P0.05).The number of positive expression of AchE neurons in intestinal myenteric plexus decreased in pneumoperitoneum 60min group compared with that in pneumoperitoneum 30min group and control group(48.00?3.16 vs 58.82?4.62,61.83?4.17,P0.05).The number of positive expression of NOS neurons in intestinal myenteric plexus increased in pneumoperitoneum 60min group compared with that in pneumoperitoneum 30min group and control group(42.17?4.45 vs 32.50?4.34,30.83?3.6,P0.05).Conclusions Prolonged CO2 pneumoperitoneum can affect or damage cholinergic neurons and nitroxidergic neurons in the enteric nervous system to some extent,and it may be the underlying mechanism of the intestinal motor dysfunction after operation.

PURPOSE: With the process of neoangiogenesis being linked to the growth and metastasis of various tumors, anticancer therapeutics with a basis in the suppression of neoangiogenesis has recently been receiving attention. In this study, we tried to clarify the immunoreactivities of vascular endothelial growth factor (VEGF), major angiogenic inducer and thrombospondin-1 (TSP-1), major angiogenic inhibitor in human Wilms' tumor and its clinicopathological significance. MATERAILS AND METHODS: Utilizing immunohistochemical staining, we assessed the immunoreactivities of VEGF and TSP-1 in archival tissues of 29 Wilms' tumors and 25 normal kidneys. Also, we assessed the relationship between expression of each factor and clinicopathological parameters in 29 cases of Wilms' tumors. RESULTS: Immunoreactivities of VEGF and TSP-1 were detected mainly in the cytoplasm of the tubular cells in normal kidneys. In Wilms' tumors, whereas VEGF was detected in the cytoplasm of the tumor cells and peritumoral stromal tissues, but TSP-1 only in the peritumoral stromal tissues. Immunohistochemical expression patterns of each factor were divided into two groups according to the area of immunoreactivity (negative:<10%, positive: > OR =10%). VEGF immunoreactivity was detected in 25 (100%) normal kidneys and in 20 (69%) Wilms' tumors. However, TSP-1 immunoreactivity was detected in 24 (97%) normal kidneys and in 3 (10%) Wilms' tumors. Therefore, although no significant difference was observed between the expressions of VEGF and TSP-1 in normal kidney, the TSP-1 immunoreactivity was significantly lower than VEGF immunoreactivity in Wilms' tumors. A relatively higher rate of positive expression of TSP-1 was observed in the patients with no demonstrable lymph node metastasis. Also, as for the VEGF, maximal diameter of the tumor was larger in the positive expression group. However, it proved otherwise for TSP-1 as the negative expression group demonstrated tumors with larger maximal diameters. CONCLUSIONS: Our study demonstrated that the TSP-1 immunoreactivity was significantly lower than VEGF immunoreactivity in Wilms' tumors, and disease progression has a tendency to be found in the VEGF-positive cases and TSP-1 negative cases. We suggest that the growth and metastasis of Wilms' tumor may be influenced mainly by TSP-1 decrease rather than VEGF increase.
Cytoplasm ; Disease Progression ; Humans ; Kidney ; Lymph Nodes ; Neoplasm Metastasis ; Thrombospondin 1 ; Vascular Endothelial Growth Factor A* ; Wilms Tumor*

Objective To establish an early cognitive disorder model in rats and investigate the early cognitive functioning after ischemic hypoxic brain injury during the neonatal period. Methods Forty-six newborn Sprague-Dawley rats were randomized into a 21-d-old group and a 31-d-old group. These 2 groups were then subdivided into model and sham-operated subgroups (M21, n=12; SH21, n=11; M31, n=12; SH31, n=11). A model of neonatal early cognitive disorder was established in the rats of the M21 and M31 groups using a modification of Rice's method. Rats in the SH21 and SH31 groups received skin incisions and common carotid artery separation without ligation or hypoxia. Each group was tested with a Morris water maze. The rats were sacrificed after testing, and brain tissue was examined under the electron microscope. Nissl staining allowed Nissl body quantification and neurocyte acin the M21 group was significantly longer than in the SH21 group. The 31-d-old subgroups had shorter average escaping latencies than the corresponding 21-d-old subgroups. (b) Spatial memory: The average platform times, Ⅰ region times and Ⅰ region distances showed no significant differences among groups. ②Brain pathology (a) Gross appearance: Obvious ischemic hemisphere atrophy was observed in the M group, and no abnormality was observed in the SH group. (b) Electron microscopic observation: In the SH group cell ultrastructures in the ischemic hippocampus were normal. Karyopyknosis and dilated endoplasmic reticulums were found in the M group. More mitochondria were found in the presynaptic membranes of the ischemic hippocampus in the M group than that in the SH group. (c) Nissl body quantification and neurocyte activity analysis: Significantly less activity in the ischemic cortex was found in the M21 group compared to the SH21 group. More activity was observed in the 31-d-old subgroups than in the corresponding 21-d-old subgroups. Conclusions ①The neonatal rats with ischemic hypoxic brain injury had prolonged average escaping latency and depressed neuronal activity. ②The 31-d-old rats had better spatial localization learning ability than the 21-d-old rats.

AIM:To study the effect of lutein intervention on visual function of patients with early age-related macular degeneration (AMD).METHODS:Totally 200 early AMD patients were divided into lutein intervention group (20mg/d) and placebo group by a randomized,double-blind,placebo-controlled trail.Questionnaire investigation,serum lutein concentration and visual function were conducted at baseline,12,24,36 and 48wk respectively.RESULTS:The serum lutein concentration in lutein intervention group was higher than the baseline (P＜0.05) and those of placebo group at 12,24,36 and 48wk after intervention (P＜ 0.05).The spatial frequencies of contrast sensitivity (CS) at low (3 cycles/degree) and medium (6 cycles/degree) in lutein intervention group,macular pigment optical density (MPOD) and best-spectacle corrected visual acuity (BCVA) were higher than those of placebo group (P＜ 0.05).Contrasted to baseline,the low and medium spatial frequencies of CS in lutein intervention group were higher (P＜ 0.05),while,there were no statistical significance in MPOD and BCVA between before and after treatment (P＞0.05).CONCLUSION:Lutein intervention can improve the visual function of patients with early AMD.

OBJECTIVETo investigate allelic frequencies of interluekin-10 (IL-10) gene promoter in Miao, Dong and Buyi ethnics of Guizhou.

METHODSTaqMan MGB-based real-time PCR was used to determine the genotypes of IL-10 -819 and IL-10 -592 in 589 Miao, Dong and Buyi ethnics of Guizhou.

RESULTSThe allelic frequency of IL-10 -819 in Miao ethnics was significantly different from those in Dong or Buyi ethnics. Allelic frequencies of IL-10 -592 in Miao ethnics was significantly different from those in Dong or Buyi ethnics. In Miao, Dong and Buyi ethnics, the distributions of genotype frequencies of IL-10 -819 and IL-10 -592 were statistically different from Han ethnics from Guizhou and Taiwan of China as well as South Koreans.

CONCLUSIONThere is a heterogeneity in the frequencies of polymorphisms of IL-10 promoter among different ethnic groups.

Alleles ; Asian Continental Ancestry Group ; ethnology ; genetics ; China ; ethnology ; Gene Frequency ; Genetics, Population ; Genotype ; Humans ; Interleukin-10 ; genetics ; Polymorphism, Single Nucleotide ; Population Groups ; genetics ; Promoter Regions, Genetic

OBJECTIVETo study the frequency of a 9 bp deletion polymorphism of mitochondrial DNA (mtDNA) in ethnic Miao, Buyi and Dong populations from Guizhou province.

METHODSPolymerase chain reaction-polyacrylamide gel electrophoresis (PCR-PAGE) was used to detect the 9 bp deletion. The result was verified with DNA sequencing.

RESULTSTwo polymorphisms, including a standard pattern and a short pattern (the 9 bp deletion), were found among the three ethnic groups. The frequency of short pattern in 304 males was 23.0%. Respectively, those of Miao, Buyi and Dong ethnics were 28.6%, 26.8% and 13.7%. A statistically significant difference was detected among the three groups (P<0.05).

CONCLUSIONThe frequencies of the 9 bp polymorphism were relatively high among ethnic Miao, Buyi and Dong populations from Guizhou, and there was a significant difference between the three.

Base Sequence ; China ; ethnology ; DNA, Mitochondrial ; genetics ; Gene Deletion ; Humans ; Male ; Molecular Sequence Data ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Sequence Analysis, DNA

OBJECTIVETo understand the function of Sonic hedgehog in chiasm development in mouse embryos of embryonic day 13 (E13) to E15.

METHODSBrain slices of E13-E15 mouse embryos containing the optic pathway from the eyes to the optic tract were prepared and cultured in DMEM/F12 in the presence of 10% fetal bovine serum at 37 degrees in a rolling incubator for 5 h. The antibody to Shh was added into the culture medium of the slices in the treatment group, while no additional chemical or only normal mouse IgG was added in the control groups. After culture, the brain slices were fixed and a DiI granule was inserted into the optic disc in one eye. Seven days later, the tissue overlying the chiasm was removed to expose the DiI-labeled chiasm for observation under confocal microscope, and the images were analyzed by METAMORPH software.

RESULTSShh antibody treatment produced a reduction of crossing of the earliest retinal axons at the midline of E13 chiasm, and the uncrossed axons were also influenced by Shh antibody at E15.

CONCLUSIONShh executes a transient but important function in axon decussation in the early stage of mouse optic chiasm development and signals axon turning in the later stage.

Animals ; Antibodies ; immunology ; metabolism ; Hedgehog Proteins ; immunology ; metabolism ; Mice ; Neural Pathways ; embryology ; metabolism ; Optic Chiasm ; embryology ; metabolism ; Retina ; embryology ; metabolism ; Signal Transduction ; Tissue Culture Techniques

Objective To establish a method for determination of the content of Forsythoside A in Yinqiaosan granules . Methods HPLC method was adopted to carry on the determination .Welch Ultimate XB‐C18 column (250 mm × 4 .6 mm ,5μm) was used with the mobile phase of methanol‐0 .2% Glacial acetic acid solution by gradient elution . The flow rate was 1 .0 ml/min ,the detection wavelength was 330 nm ,and the column temperature was 20 ℃ .Results Forsythoside A showed a good linear relationship at the range from 0 .080 3~3 .211 μg (r=1 .000) .The average recovery rates were 103 .78% ,and RSD=0 .82% .Conclusion The method was convenient and accurate .Meanwhile ,it also has good specificity and reproducibili‐ty .It can be used effectively for the quality control of Yinqiaosan granules .

Objective To study the genotype distribution of insulin receptor substrate-2(IRS-2)gene 1057G/A polymorphism in Han population from Southwest China,and to explore its association with the metabolism of glucose and lipids,insulin resistance and islet?-cell function in type 2 diabetic patients and subjects with impaired glucose tolerance(IGT).Methods A total of 929 Hans[462 patients with type 2 diabetes(DM group) and 164 subjects with IGT(IGT group)and 303 normal controls(NC group)]from Chongqing and nearby regions were screened for 1057G/A polymorphism of IRS-2 gene by PCR-RFLP assay.Body mass index(BMI),plasma glucose,serum insulin and lipid profile,high-sensitive C-reactive protein(hsCRP)and non-esterified fatty acid were measured.Homeostasis model assessment of insulin resistance(HOMA-IR)and disposition index(DI)were used to estimate insulin resistance and?-cell function respectively.Results In DM group,A allele frequency was significantly lower than that in NC group(0.326 vs 0.388,X~2=6.19,P=0.01).Compared with NC group,AA genotype frequeney was lower and GG genotype frequeney was higher in DM group(0.104 vs 0.135 and 0.452 vs 0.360 respectively,X~2=6.80,P