Adult ; Aged ; Carbon Monoxide Poisoning ; blood ; pathology ; Creatine Kinase ; blood ; Female ; Humans ; Male ; Middle Aged ; Myocardium ; pathology ; Natriuretic Peptide, Brain ; blood ; Peptide Fragments ; blood ; Troponin I ; blood ; Young Adult

Objective To investigate the influence of fixation of both lower limbs with negative pressure vacuum cushion and fixation of both ankles with self-made foam mat on setup errors in radiotherapy for rectal cancer.Methods A total of 12 patients with rectal cancer were enrolled in 2014 and randomly divided into group A (using negative pressure vacuum cushion) and group B (using self-made foam mat).An offline registration analysis was performed for the images of 108 times (A,B group of 54 times) of kilovoltage cone-beam CT (CBCT) before and after treatment.Grey scale translation error registration was used,and the results of registration were analyzed.The setup errors in x-axis (left-right direction),y-axis (cranial-caudal direction),and z-axis (anterior-posterior direction) were compared between the two groups.Results There was no significant difference in the absolute setup error in the y-axis between the two groups (2.13±0.64 mm vs.2.61±1.17 mm,P=0.399),while group A showed significantly lower absolute setup errors in the x-axis and z-axis than group B (x-axis:1.51±0.28 mm vs.2.70±1.05 mm,P=0.039;with an error rate of 7.41％ vs.42.59％;z-axis:1.10±0.29 mm vs.2.37±0.71 mm,P=0.002;with an error rate of 1.85％ vs.35.19％).Conclusions In the radiotherapy positioning for rectal cancer,fixation of both lower limbs with negative pressure vacuum cushion effectively avoids the translation and rotation of both lower limbs,reduces absolute setup errors,and has higher accuracy than fixation of both ankles with self-made foam mat.

Background and purpose:Ovarian cancer-associated ifbroblasts (CAF) are known to promote epithelial malignancy. The chemoattractant cytokine growth-regulated oncogene alpha (Gro-α) secreted from CAF has been reported to mediate the stroma-epithelia interaction in tumor microenvironment, leading to the development of epithelial ovarian cancer, however, the detailed mechanism is unknown.This study was to determine whether Gro-αcould promote ovarian tumorigenesis through activating NF-кB nuclear translocation and VEGF expression in stromal ifbroblasts. Methods:ELISA was used to measure the levels of Gro-αin two cancer-associated ifbroblasts (CAF) and normal ifbroblasts (NF) isolated from high-grade serous ovarian cancer or normal ovarian tissues. CAF conditioned medium (CM) or Gro-αwas used to treat NF, while PS1145, the inhibitor of NF-кB, was used as control. NF-кB subunit p65 and vascular endothelial growth factor (VEGF) were detected by Western blot in cells after treatment. Xenograft tumors from nude mice were generated by injection of CAF, NF, or OVCA429 alone or OVCA429 mixed with CAF or NF, and by injection of OVCA429 mixed with NF cells that were treated with or without CAF-CM or Gro-α, or with NF cells that were treated with CAF-CM or Gro-αplus PS1145. The tumor growth curve was measured and the blood vessel density in xenograft tumor tissues was examined by histopathological analysis. Results:The levels of Gro-αwere 5-6 folds higher in CAF than in NF. Treatment of NF with CAF-CM or Gro-αstimulated the nuclear translocation of NF-кB subunit p65, and the expression of VEGF, but suppressed the expression of thrombospondin 1, the anti-angiogenesis factor, compared with control cells. However, treatment of NF with the NF-кB inhibitor PS1145 reversed these results. The animal assay revealed that CAF stimulated tumor growth stronger than NF, and NF treated with CAF-CM or Gro-α, but not along with PS1145, enhanced xenograft tumor growth through promoting angiogenesis. Conclusion:Ovarian CAF promotes the nuclear translocation of NF-кB and the expression of VEGF through Gro-αautocrine in tumor microenvironment to facilitate angiogenesis and ovarian cancer development.

Background and purpose: Cancer-associated fibroblasts (CAFs) are known to promote the invasion and metastasis of epithelial cancers. The cytokine IL-6 may mediate the interaction between stromal cells and epithelia in tumor microenvironment to facilitate the invasiveness and metastasis of cancer, however, such mechanism has not been fully covered yet.Methods:We used cervical cancer cell line HeLa as a model for this study. ELISA was used to measure the levels of IL-6 in CAFs and normal ifbroblasts (NFs) isolated from squamous cervical cancer or normal cervical tissues. CAFs conditioned medium or IL-6 was used to treat cervical cancer HeLa cell line. The epithelial-mesenchymal transition (EMT) markers such as N-Cadherin and Vimentin were detected by Western blot in cells before and after treatment. Scratches and transwell chambers were used to test the abilities of cell migration and invasion. Results:The levels of IL-6 were 4-5 folds higher in CAFs than in NFs. Treatment of HeLa cells with CAF conditioned medium or IL-6 upregulated N-Cadherin and Vimentin, but down-regulated E-Cadherin and cytokeratin, compared with control cells, indicating that IL-6 may stimulate HeLa cells to EMT. Further study found that Snail 1, the featured transcription factor for stem cells, was increased along with the enhanced phosphorylation of STAT3. Meanwhile, the migration and invasion of HeLa cells treated with IL-6 or CAF conditioned medium were markedly increased. Conclusion:CAF induces the EMT of cervical epithelial cancer cells through IL-6/STAT3/Snail pathway, which thereby promotes the invasiveness and metastasis cervical epithelial cancer.

A reporter system for φC31 integrase was developed in NIH3T3 cells. The reporter plasmid coding green fluorescent protein (GFP) coupled with red fluorescent protein (RFP) was eo-transfected with the plasmid coding φC31 integrase, to show the activity of integrase in the cells. Fluorescence activated cell sorter (FACS) was used to measure the proportion of the cells containing red and green fluorescence. The increment of green cells was positively related to the increase in the transfection with plasmid coding φC31 integrase. Approximately 90% of green cells were observed under a ratio of [plasmid-φC31-integrase]/[reporter plasmid] at 10 : 1. This suggests that the φC31 integrase reporter system provides a probe for the function of φC31 integrase in cells.

Objective To study the value of CT on defining the gross tumor volume (GTV) in three-dimensional conformal radiotherapy (3DCRT) and intensity-modulated radiation therapy(IMRT), by comparing the GTV contoured on preoperative CT with postoperative pathology guided GTV in patients with non-small cell lung cancer (NSCLC). Methods Thirty-three patients with histologically proved NSCLC had had CT scan of the thorax before sugery. We contoured the GTV for both primary tumor (GTV-T) and the regional metastatic lymph nodes (GTV-N) on preoperative CT scan. The GTV-T was defined on lung window while the GTV-N on mediastinal window. The metastatic lymph node was defined as≥ 10mm in diameter of short axis,while

Objective:To study the expressione of autophagy-related protein Beclin-1 and LC3 in cortex and hippocampus of rats after cerebral ischemia reperfusion injury and the efficacy of Edaravone.Methods:Sprague Dawley rats were randomly divided into sham group,model group and Edaravone group.The cerebral ischemia reperfusion model was induced via Zea Longa with blocking the middle cerebral artery of 2 h and reperfusing of 24 h.Animals assigned to sham group were only separated left common carotid artery.Edaravone was injected intraperitoneally at a dose of 1 0 mg/kg at 15 min before reperfusion.The condition of nerve injury of rats was conducted by Neurobehavioral score.The degree of brain injury and success of model were determined based on 2,3,5-triphenyltetrazolium chloride(TTC)staining.The changes of neuron stained by hematoxylin and eosin (HE) in cortex and hippocampus were observed.The expression of Beclin-1 and LC3 was measured by immunohistochemistry.Results:After cerebral ischemia reperfusion the neurobehavioral score of edaravone group was(2.00± 0.67),which was obcviously less than(2.50± 0.53) of model group(P＜0.05).The infraction focus and the neuron injury in cortex and hippocampus neurons were also observed in model group,and the edaravone group reduced above expression.The positive rate of Beclin-1 of each group in cortex were (1 1.08± 0.85)％,(33.42± 1.57)％ and (25.61± 1.39)％,there was significant difference between model group with sham group and edaravone group (P＜0.05).The positive rate of Beclin-1 of each group in hippocampus were (10.34± 0.21)％,(31.82± 1.73)％ and (22.74± 1.26)％,there was significant difference between model group with sham group and edaravone group(P ＜ 0.05).The positive rate of LC3 of each group in cortex were (15.33± 0.47)％,(39.72± 1.73)％ and (28.53± 1.61)％,there was significant difference between model group with sham group and edaravone group(P＜0.05).The positive rate of LC3 of each group in hippocampus were (13.74± 0.37)％,(32.53± 1.43)％ and(25.38± 1.23)％,there was significant difference between model group with sham group and edaravone group (P＜0.05).Conclusion:The expression of Beclin-1 and LC3 was increased after cerebral ischemia reperfusion.Edaravone may reduce autophagy and brain injury through downregulation the expression of Beclin-1 and LC3.

A reporter system for ?C31 integrase was developed in NIH3T3 cells.The reporter plasmid coding green fluorescent protein(GFP) coupled with red fluorescent protein(RFP) was co-transfected with the plasmid coding ?C31 integrase, to show the activity of integrase in the cells.Fluorescence activated cell sorter(FACS) was used to measure the proportion of the cells containing red and green fluorescence.The increment of green cells was positively related to the increase in the transfection with plasmid coding ?C31 integrase.Approximately 90% of green cells were observed under a ratio of plasmid-?C31-integrase/reporter plasmid at 10∶1.This suggests that the ?C31 integrase reporter system provides a probe for the function of ?C31 integrase in cells.

AIM: To investigate the role of p38 mitogen-activated protein kinase (p38MAPK) signaling pathway in the Kupffer cells (KCs) production of pro-inflammatory cytokines, tumor necrosis factor-?(TNF-?) and interleukin-1?(IL-1?), in severe acute pancreatitis (SAP) rats. METHODS: Sprague-Dwaley rats were randomized into three groups: ①sham operation rats, ②SAP rats, ③SAP rats given the p38 MAPK inhibitor CNI-1493(10 mg/kg, iv). The SAP model was induced by the bili-pancreatic duct infusion with 5% sterile soduim taurocholate solution. Rats from each group were killed at 12 h after sham operation or SAP and Kupffer cells (KCs) were isolated. The mRNA expressions of TNF-? and IL-1? (by quantitative real-time RT-PCR) and p38 MAPK activity (by Western blot analysis) in KCs were examined. The levels of TNF-? and IL-1? in plasma were determined by ELISA. RESULTS: There was a significant acvitation of p38 MAPK in KCs harvested from SAP rats than those from sham operation rats. SAP also promoted the mRNA expressions of TNF-? and IL-1? in KCs and the plasma levels of TNF-? and IL-1?. These events were significantly inhibited by treatment with CNI-1493.CONCLUSIONS: p38 MAPK activation is one important aspect of the signaling events that may mediate the KCs production of pro-inflammatory cytokines, TNF-? and IL-1?, in SAP rats. The inhibition of the p38 MAPK may be a potential target in the prevention and treatment of SAP.

We produced (S)-4-cyano-3-(4-chlorophenyl)-butyrate by highly stereoselective biocatalyst in this study. A nitrilase-producing strain, named Gibberella intermedia WX12, was isolated by 3-(4-chlorophenyl)-glutaronitrile as substrate in the screening with phenol-sodium hypochlorite method. The fermentation conditions and catalytic properties of this strain were investigated. The preferred carbon and nitrogen sources for nitrilase production were lactose (30 g/L) and peptone (20 g/L). After being cultivated for 96 h, the cells were collected for use in biotransformation. The hydrolysis of 3-(4-chlorophenyl)-glutaronitrile was performed at 30 degrees C in phosphate buffer (pH 8.0, 50 mmol/L) for 24 h to give (S)-4-cyano-3-(4-chlorophenyl)-butyric acid with 90% yield and > 99% of ee, which can be used for the synthesis of (R)- and (S)-baclofen. The configuration of product was determined by chemically converting it to baclofen and comparison with the authentic sample by chiral HPLC analysis.
Aminohydrolases ; metabolism ; Baclofen ; chemical synthesis ; chemistry ; Biocatalysis ; Chlorophenols ; chemistry ; Gibberella ; enzymology ; Hydrolysis ; Nitriles ; chemistry ; Prodrugs ; chemical synthesis ; chemistry