Background and purpose: The morbidity of colorectal cancer in China increased year by year. This study aimed to explore the signiifcance of ADAMTS9 protein levels and promoter methylation in colorectal cancer onset and progression. Methods:ADAMTS9 promoter methylation status was detected by methylation speciifc PCR method in 162 colorectal cancer patients’ peripheral blood DNA samples; Plasmatic ADAMTS9 protein levels was detected by enzyme-linked immunosorbent assay method in 162 colorectal cancer patients and 150 healthy subjects. Results: Compared with healthy people, patients with colorectal cancer had a significant lower ADAMTS9 protein level in plasma [(65.25±9.70)μg vs (50.28±9.66)μg, P<0.001];ADAMTS9 gene promoter methylation was observed in 66 among 162 colorectal cancer patients (40.7%);The plasma level of ADAMTS9 protein in patients with methylated ADAMTS9 gene had signiifcantly reduced (P<0.001), while the plasma level of ADAMTS9 protein in patients with low ADAMTS9 protein had signiifcantly increased (P=0.007);ADAMTS9 methylation is closely related to tumor size (larger, P=0.017) and tumor differentiation degree (P=0.029), ADAMTS9 protein low expression is closely related to invasion depth (P=0.020), lymph node metastasis (P=0.019) and Dukes staging (P=0.002).Conclusion: ADAMTS9 protein downregulation induced by DNA promoter methylation may be involved in the pathogenesis, invasion and metastasis, and promote the progression in colorectal cancer.

Objective To investigate the value of single photon emission computed tomography (CT) imaging and transmis‐sion CT imaging (microSPECT‐CT ) bremsstrahlung imaging for the solid tumor mesenchymal implantaion of 32 P‐chromic phos‐phate‐paclitaxel‐poly‐L‐lactic acid (32 P‐CP‐PSP‐PLLA) sustained release seeds and to investigate the 32 P in vivo biodistribution and degradation sustained release churacteristics .Methods The animal model of prostate cancer subcutaneously transplanted tumor was established .32 P‐CP‐PSP‐PLLA sustained‐release seeds were intratumorally implanted by the mediation of microSPECT‐CT brems‐strahlung imaging and the 32 P distribution in bearing tumor mouse was verified by the imaging and biological distrubtion tests .The ultrastructural changes of 32 P seeds were observed by the scanning electron microscope .Results The MicroSPECT/CT brems‐strahlung imaging could effectively guide the intratumoral implantation operation of the 32 P sustained‐release seeds with clear visu‐alization .Partial sustained‐release 32 P was remained in the tumor tissues with little distribution in important organs of spleen and liver ,which was proved by the biodistribution results .The particle surface and inside micropores and tunnels formation ,their pro‐gressive increase ,fusion and connection were found by the electronic microscope after the 32 P sustained‐release seeds intratumoral implantation .Conclusion The MicroSPECT/CT bremsstrahlung imaging can effectively monitor the 32 P sustained‐release seeds and their in vivo biodistribution and lays a foundation for the sustained‐release seeds prostatic targeted implantation .

Objective To study the effect of specific hammerhead ribozyme (Rz) to hepatitis C virus (HCV) in vitro. Methods Rz 1 Rz 2 were designed to cleavage at 5′-NCR nucleotide positions under 136～160 and 313～337, Rz 3 was designed to cleavage at C region nucleotide position under 373～388. As a control, cleavage deficient Rzm that have A→G point mutations in the catalytic loop of the hammerhead domain. 32P-labeled transcript of target HCV RNA was incubated with gel-purified Rz ( Rz 1, Rz 2, Rz 3 and Rzm ) respectively at different concentration based on specified condition and autoradiographed after denaturing gel-electrophoresis. Results Except Rzm, Rz 1 Rz 2 Rz 3 were active at 37 ℃ and more so at higher concentration, and more so with cleavage site nearly to the HCV initial code. Conclusions The HCV specific hammerhead ribozyme can be designed in vitro, further study about cleavage in vitro and in vivo will continue.

BACKGROUND: Previous studies have found that nerve growth factors play an important role in the process of wound healing, but there is less research for the low-affinity nerve growth factor receptor p75 and sortilin in fibroblasts, and no reports on whether there are differences in expression of p75 and sortilin in the scar fibroblasts and normal skin fibroblasts. OBJECTIVE: To study the expression of low-affility nerve growth factor receptor p75 and sortilin in the normal human skin fibroblasts and the human keloid fibroblasts. METHODS: The keloid fibroblasts and normal hunman skin fibroblasts were cultured in vitro, and the immortalized epithelial cells HaCaT were used as the positive control. The real-time PCR was used to detect the mRNA expression of the p75 and sortilin in the keloid fibroblasts and normal human skin fibroblasts, and western blot and immunocytochemical staining were used to detect the protein expression of p75 and sortilin. RESULTS AND CONCLUSION: The real-time PCR and western blot results showed that in the protein and mRNA levels, p75 and sortilin showed positive expression in the keloid fibroblasts and normal human skin fibroblasts, and there was no significant difference in the expression of p75 between keloid fibroblasts and normal human skin fibroblasts, and the expressions of p75 and sortilin in the keloid fibroblasts and normal human skin fibroblasts were significantly lower than those in HaCaT. There was no significant difference of p75 expression between keloid fibroblasts and normal human skin fibroblasts, and the expression of sortilin in the keloid fibroblasts was significantly lower than that in the normal human skin fibroblasts (P < 0.05). Immunocytochemical staining result showed that the expression of p75 and sortilin in the keloid fibroblasts and normal human skin fibroblasts were distributed in the membrane and cytoplasm. Precursor nerve growth factor combined with high-affinity p75 receptor could promote the apoptosis of the cells with the help of sortilin, and the expression of sortilin in the keloid fibroblasts was significantly lower than that in the normal human skin fibroblasts, which may associated with the high proliferation of the keloid fibroblasts. The results provide a new target for the prevention and treatment of pathological scars.

Amino Acids ; urine ; Animals ; Biogenic Monoamines ; metabolism ; urine ; Male ; Microwaves ; Rats ; Rats, Wistar

Endophthalmitis ; diagnosis ; drug therapy ; etiology ; Eye Infections, Fungal ; diagnosis ; drug therapy ; etiology ; Humans ; Infant, Newborn ; Infant, Premature ; Male

Objective To investigate the feasibility and value of CT perfusion imaging in combination of CTA 320-slice CT with dual-low-dose in detection of acute cerebral ischemia.Methods Forty patients with acute stroke underwent perfusion imaging and CTA using a single scan with 320-slice CT with dual-low-dose schedule.CTA,4D-CTA,4D-perfusion imaging and fusion images were generated at a post-processing workstation.All patients also underwent a 3.0T magnetic resonance imaging with DWI for a comparison.Quality of the images,degree of vascular stenosis,and location of ischemic lesions were evaluated.Results The vascu-lar stenosis or occlusion was found in 33 patients,8 of whom were confirmed by the DSA.The ratio of better images of CTA was 82.5%.CTP showed 297 lesions with abnormal perfusion.202 lesions were confirmed as infarct by DWI,while 95 ones were nor-mal on DWI but with increased DLY and TTP and slightly decreased CBF and CBV in 49,increased DLY and TTP and normal CBF and CBV in 21,and increased DLY and TTP and slightly increased CBF and CBV in 25.Conclusion CT perfusion in combination with CTA by 320-slice CT with dual-low-dose is feasible and useful to observe accurately the vascular structures and the ischemic status of the whole brain at early stage.

ObjectiveTo investigate the effect of immunoglobulin G (IgG) extravasated from blood circulation on the expression of toll-like receptor 4 (TLR4) induced by peripheral lipopolysaccharide (LPS) in rat brain. Methods The rats were divided into four groups in random, 5 rats in each. Group one received LPS 100μg/kg by intraperitoneal administration, normal saline was given by intravenous injection 6 hours later; group two was injected with adrenalin (AD) 15μg/kg intravenously; group three was treated with LPS intraperitoneally, AD was injected 6 hours later; group four was injected normal saline intravenously as control. For all groups, the animals were sacrificed 30 min after the last injection, and the brains were taken for investigation of the TLR4 expressions by immunofluorescence staining and RT-PCR. Result Immunofluorescence staining showed that IgG immunoreactive product was patch-like, distributed in the brain parenchyma in all the animals that received AD. In the LPS+normal saline group, IgG was found merely around the blood vessels. Meanwhile, in LPS+AD animals, TLR4 immunoreactive product coexisted with microglia marker Iba-1 within the IgG extravasated area. The double-labeled cells dispersed in the brain parenchyma and near to the cerebral vessels. In the LPS+saline group, TLR4 positive cells were endothelial-like. RT-PCR results indicated that the expression level of TLR4 in the LPS+AD group were significantly higher than that in the LPS+saline group or AD group or the saline control (P<0.01). Conclusion Extravasated circulating IgG may enhance the TLR4 expression in the rat brain induced by peripheral LPS.

Objective Study the relationship between HLA antibodies production and the number of Gravidity.Methods A total of 856 pregnant women of first pretanal in Xiasha Hospital of Sir Run Run Shaw Hospital of Zhejiang University from Nov 2016 to Mar 2017 were recruited.The blood samples were collected during 24-28 weeks of pregnancy,and the expression of platelet-specific alloantibody was detected by solid-phase red cell adherence assay.Results There were 67 positive HLA antibody in pregnant women,and the antibody positive rate was 7.8％.There were 15 positive HLA antibody in 334 first pregnant women,14 in 214 two or more pregnancy pregnant women without abortion,10 in 107 pregnant women with once abortion,28 in 223 pregnant women with two or more abortion.The antibody positive rate respectively is 4.5％,6.5％,9.3％ and 12.6％.The production of HLA antibody was significantly associated with the numberof gravidity.Conclusion The antibody positive rate of two or more pregnancy is significantly higher than the fisrt pregnancy.