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Objective To evaluate the imaging characteristics of atypical teratoid/rhab doid tumor (AT/RT) of central nervous system(CNS), and to improve the diagnostic ability of the disease. Methods The clinical and imaging findings of 9 patients were retrospectively analyzed. There were 5 male and 4 female, ages 7 months to 5 years,median age was 1.4 years. MR enhancement studies were obtained in all the cases. One case had CT enhancement examination. Results The lesions were seen in brain in 8 cases and in lumbosacral spinal cord in one case. The tumors size varied from 4.8—7.8 cm, Necrosis was seen in nine cases, cystic change in eight cases and hemorrhage in five cases. The tumors had high signal on DWI, and low signal on ADC map. Dura matter invasion(2 cases), cerebrospinal fluid spread(2 cases)and intracerebral metastasis were seen. Conclusion There are some relatively specific imaging findings of primary CNS AT/RT that could assist their diagnosis.
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To monitor genetic variation of porcine reproductive and respiratory syndrome virus (PRRSV),RT-PCR was used to identify a sample suspected of PRRSV infection.A PRRSV named SC-GY strain was obtained,and its Nsp2,ORF5 and ORF3 genes were used for sequence alignment and phylogenetic tree construction.The results showed that SC-GY strain is highly pathogenic PRRSV American variant strains with Nsp2 gene discontinuous deletion of 30 amino acids,ORF3 gene aa17 a serine (S) insert.Comparing to VR2332,CH-1a,JXA1,HUN4,NADC30,HENAN-XINX and SC2012,the Nsp2,ORF5 and ORF3 of SC-GY shared 70.3％-97.9％,82.4％-97.6％ and 83.1％-98.2％ of nucleotide similarity,and 62.3％-96.3％,78.0％-95.7％ and 81.6％-96.5％ of deduced amino acid similarity;and compared to LV they shared only 18.9％,60.8％ and 63.7％ of nucleotide similarity,and 14.0％,54.9％ and 57.2％ of deduced amino acid similarity.The phylogenetic tree revealed that the SC-GY formed independent small branches although it belonged to the same subgroup as highly pathogenic PRRSV strains.The results showed that in high frequency live vaccine immunization of currently PRRSV,the gene of PRRSV epidemic strain is still in constant variation.Vaccination of live PRRSV vaccines should be reduced and surveillance of PRRSV strains should be enhanced.
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<p><b>OBJECTIVE</b>To investigate the association of gankyrin protein expression in colorectal cancer (CRC) with its prognosis.</p><p><b>METHODS</b>Clinical data and resection samples of 100 colorectal cancer patients identified by pathology undergoing resection in our department from June 2008 to June 2009 were collected. The gankyrin expression in CRC tissues and matched adjacent noncancerous tissues collected during the operation of 100 CRC cases was detected by immunohistochemical staining and Western blotting. The associations of gankyrin expression level with overall survival, clinicopathologic features were analyzed by Chi square test, Cox regression analysis, Kaplan-Meier analysis and log rank test.</p><p><b>RESULTS</b>Immunohistochemical staining showed that the positive brown granules were mainly distributed in the cytoplasm, and nuclear immunostaining was observed in tissue samples of 29 cases, of whom 16 cases had distal metastasis [55.2% (16/29)]. The positive rate of gankyrin and the relative gray value of Western blotting in CRC tissues were 67% (67/100) and 0.69±0.23, respectively, which were significantly higher than those of 2 cm adjacent noncancerous tissues [6% (6/100) and 0.31±0.16] and 10 cm adjacent noncancerous tissues [1%(1/100) and 0.16±0.11] (all P<0.001). Patients with positive expression of gankyrin had worse survival than those with negative ones (41.8% vs. 72.7%, P=0.008). The gankyrin expression was associated to lymph node metastasis (P=0.005), tumor stage (P=0.001) and distal metastasis (P=0.002). Cox regression analysis showed that distal metastasis (P=0.004) and high expression of gankyrin (P=0.038) were independent risk factors for poor prognosis of patients with CRC.</p><p><b>CONCLUSION</b>Up-regulated expression of gankyrin is related to invasion and metastasis of human CRC, and gankyrin may be valuable in predicting prognosis.</p>
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Humans , Blotting, Western , Colorectal Neoplasms , Kaplan-Meier Estimate , Lymphatic Metastasis , Postoperative Period , Prognosis , Proteasome Endopeptidase Complex , Proto-Oncogene ProteinsABSTRACT
PurposePrimary chest wall rhabdomyosarcoma (RMS) is very rare with limited imaging characteristic studies in the literature. This paper analyzes the CT imaging features of chest wall RMS in children to improve the diagnostic accuracy.Materials and MethodsThe imaging data of contrast enhanced CT scan of pathology conifrmed chest wall RMS in ifve children were analyzed.ResultsThe lesion was located in the anterior chest wall in one case, in the posterior chest wall in two cases, and the lateral chest wall in two cases (axillary). The tumors were round or spindle in shape with shallow spiculation. Plain CT showed heterogeneous density with patchy low-density necrotic area in two cases, and homogeneous attenuation in three patients. In all ifve cases there was no calciifcation or fatty tissue. The tumor involvement of adjacent spinal canal was seen in one case. Visceral compression was evident including lung parenchyma in one case, heart and liver in one case. Tumor blood vessel growth was seen in two cases. All ifve lesions were adjacent to the ribs, humerus, scapula and the spine with bone destruction in one case. On contrast enhanced scan, all ifve cases demonstrated heterogeneous mild to moderate enhancement, more prominent in the periphery. There were enlarged feeding arteries. Necrotic areas did not enhance. In two cases there were pulmonary metastases. Pleural effusion and ascites were identiifed in one case. There was lymph node metastasis in one case.ConclusionThe CT manifestation of children's chest wall RMS for chest wall include large soft tissue mass, heterogeneous density, no calciifcation or fatty tissue, partial necrosis, adjacent tissue compression, lymph node or distant metastasis. Combining with clinical manifestations, comprehensive analysis of contrast enhanced CT imaging can improve diagnostic accuracy.
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Objective To investigate the micro-vessel density and vascular endothelial growth factor receptor 2 (VEGFR2) expression in mucosa and submucosa tissue of the patients with hemorrhoids. Methods Sixty hemorrhoids patients were randomly recruited and involved into experiment group and 20 cases of normal patients involved in control group. RASP, MASP, RSTV, RMTV, Expression of VEGFR2 and microvessel density in two groups were detected by immunohistochemistry. The expression abundance of VEGFR2 mRNA was detected by RT-PCR. Results Excessive angiogenesis was found in hemorrhoids tissue (5.59 ± 0.98) in experiment group as compared with that in control group (3.92 ± 0.81), the difference was statistically significant (P < 0.05). Staining score of hemorrhoid tissue and the control tissue were 5.95 ± 1.01, 2.32 ± 0.72, respectively (P < 0.05). Abundance of VEGFR2 mRNA expression in hemorrhoid tissue was significantly higher than that in control tissue (P < 0.05). Conclusion VEGFR2 plays an important role in angiogenesis during the development of hemorrhoids, which may be closely related to the occurrence of hemorrhoid disease and clinical symptoms.
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Objective To investigate the effects of gambogic acid on proliferation,invasion and protein expression of MMP-2 in LoVo human colon cancer cell line.Methods The CCK-8 assay was used to determine the anti-proliferation ability.Transwell chamber assay was used to study the invasion of LoVo colon cancer cells.Western Blot was applied to examine the protein expression of MMP-2.Results CCK-8 assay showed that after cells treated with 1,2,4 μmol/L GA for 24 h,48 h the inhibition rates were(0.16±0.11)％,(0.24±0.08)％,(0.58±0.01)％,(0.67±0.03)％,(0.79±0.01)％ and (0.27±0.05)％,(0.69±0.09)％,(0.85±0.01)％,(0.87±0.01)％,(0.89±0.01)％,and there had statistically significant differences between control group (0 μmo/L) with experimental group (P＜0.05).Gambogic acid can effectively inhibit LoVo cells invasion at a low-dose concentration,which the numbers of cells that digested the matrigel in control group (0 μmo/L)was 120.50± 8.69 and 69.83 ±4.75 in experimental group (1 μnol/L),P＜ 0.05).Western Blot revealed that Gambogic acid can down-regulate the expression of MMP-2,which the levels of down-regulating the expression of MMP-2 after treated with 1,2,4 umol/L GA were 48.67％,74.72％and 82.70％ (P＜0.05) Conclusion Gambogic acid can effectively inhibit the growth and invasion of the LoVo cells,which may be related with down-regulating the expression of MMP-2.
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Developing a high-throughput screening method is of great importance for directed evolution of atrazine chlorohydrolase. A mutagenesis library of atzA from Pseudomonas sp. ADP and Arthrobacter sp. AD1 was constructed using error-prone PCR and DNA shuffling. Candidate mutants were screened through Haematococcus pluvialis expression system, using atrazine as selection pressure. Sequence analysis showed that mutations in the obtained 12 mutants with enhanced activity were all point-substitutions and scattered throughout the gene. Enzymatic activity analysis showed that the mutants all had higher activities than that of the wild type. The activities were 1.8-3.6 fold of the wild-type enzyme when cultured in BBM medium with 1 mg/L atrazine, whereas 1.8-2.6 fold with 2 mg/L atrazine. These results indicated that Haematococcus pluvialis expression system is an ideal high throughput screening system for directed evolution of atrazine chlorohydrolase.
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Amidohydrolases , Genetics , Atrazine , Metabolism , Bacterial Proteins , Genetics , Biodegradation, Environmental , Chlorophyta , Genetics , Metabolism , Herbicides , Metabolism , High-Throughput Screening Assays , Hydrolases , Genetics , Mutagenesis, Insertional , Pseudomonas , GeneticsABSTRACT
The Arabidopsis thaliana glutathione S-transferases zeta class (AtGSTZ) is a multi-functional enzyme, which plays important role in cellular metabolism and environmental purification. Error-prone PCR and cycles of DNA shuffling were used to construct a mutagenesis library of AtGSTZ. The screening of the resultant libraries was carried out by a pH indicator dye-based colorimetric assay. Nine mutants which enhanced the dichloroacetic acid dechlorination activity were obtained. Among them, NN23 contained 25 amino acid substitutions with the activity improving 120%, whereas NN20 contained 24 amino acid substitutions with the activity improving 102%. EC1 contained 2 amino acid substitutions with the activity improving 47%. The rest 6 mutants contained one amino acid substitution with their activity increasing from 9% to 60%. The enzymatic characterization showed that all the evolved enzymes increased their catalytic efficiencies towards dichloroacetic acid and binding affinity towards glutathione whereas some of them increased the renaturability. However there is no obvious change in their thermostability. Based on these data, functional residues related to catalysis and refolding of AtGSTZ were discussed.
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AIM: To investigate the effect of puerarin (Pur) on expression of Fas/FasL mRNA in lung tissue during pulmonary ischemia and reperfusion injury(PIRI) in rabbits.METHODS: Single lung ischemia and reperfusion animal model was used.The rabbits were randomly divided into three groups,sham operated group(sham,n=10),PIR group(I-R,n=30) and PIR+ Pur group(Pur,n=30).Changes of several parameters included apoptotic index(AI),wet to dry ratio of lung tissue weight(W/D) and index of quantitative assessment of histologic lung injury(IQA) were measured at 60,180 and 300 minutes after reperfusion in lung tissue.Meanwhile,the location and expression of Fas/FasL mRNA were observed.Lung tissue was prepared for light microscopic and electron microscopic observation at 60,180,300 minutes after reperfusion.RESULTS: As compared with group I-R,Fas/FasL mRNA slightly expressed in intima and extima of small pulmonary artery,alveoli,and bronchiole epithelia in group Pur.The values of AI,W/D and IQA showed significantly lower than that in group I-R at 60,180,300 minutes after reperfusion in lung tissue(P
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AIM: To investigate the variations of heme oxygenase-1/carbon monoxide system in lung ischemia-reperfusion injury and effects of puerarin on the system.METHODS: The unilateral lung ischemia-reperfusion model was replicated in vivo.Rabbits were randomly divided into three groups(n=10 in each): control group,ischemia-reperfusion(I-R) group and puerarin group.The blood specimens collected at times before ischemia,ischemia 1 h,reperfusion 1 h,3 h and 5 h were tested for the contents of carboxyhemoglobin(COHb) and cyclic guanosine monophosphate(cGMP).The lung tissues sampled at 5 h after reperfusion were assayed for wet/dry weight ratio(W/T),the injured alveoli rate(IAR) and the activity of HO-1.The changes of ultrastructure were observed under electron microscope.The tissue slides were also stained by immunohistochemistry(IHC) and in situ hybridization(ISH) to detect the expression of HO-1.RESULTS: The plasma content of COHb and cGMP in I-R and puerarin group increased in a time-dependent manner after I-R compared with control group,but the increment in puerarin group was higher than that in I-R group(P
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AIM: AIM: To explore the relationship between apoptosis in the lung tissues and lung ischemia/reperfusion injury, and observe effects of panax notoginseng saponins (PNS) on apoptosis in lung ischemia/reperfusion injury. METHODS: Single lung in situ ischemia/reperfusion animal model was used. Eighty four Japanese white rabbits were randomly divided into control group (control), ischemia/reperfusion 1 h group (IR1h), IR3h, IR5h, Panax Notoginseng Saponins 1 h group (PNS1h), PNS3h and PNS5h. TUNEL, immunocytochemistry and in situ hybridization techniques were used to observe apoptosis and Fas/FasL expression in various phases of lung ischemia/reperfusion. RESULTS: Cell apoptosis in lung tissues were significantly high, Fas/FasL mRNA and its protein were up-regulated in lung tissues of lung ischemia/reperfusion injury compared with control (all of P
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AIM: To investigate the effect of heme oxygenase-1 (HO-1)/carbon monoxide (CO) system on pulmonary ischemia-reperfusion injury (PIRI) in rabbits. METHODS: Single lung ischemia and reperfusion animal model was used in vivo. The rabbits were randomly divided into three groups (n=10 in each), control group (C), PIR group (I-R), PIR+ hemin group (H) and PIR+zinc protoporphyrin IX (ZnPP) group (Z). Changes of several parameters which included plasma carboxyhemoglobin (COHb) at different time points, wet to dry ratio of lung tissue weight (W/D), the injured alveoli rate (IAR) and the HO-1 enzymatic activity were measured at 180 min after reperfusion in lung tissue. The tissue slides were also stained by immunohistochemistry (IHC) and in situ hybridization (ISH) for HO-1 to detect the expression of HO-1 in lung and to analyze the optical density. The lung tissue was prepared for electron microscopic observation at 180 min after reperfusion. RESULTS: The plasma content of COHb in I-R, H, and Z group increased in a time-dependent manner after I-R. But the increment of H group was higher than that of I-R group, while that of Z group was lower. The HO-1 activity in lung tissue was highest in H group, followed by IR group, Z group, and C group (P
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Objective To evaluate the diagnosis and treatment of severe pancreatic trauma(SPT). Methods The clinical data of 32 patients with transected pancreatic trauma(grade 3-5) admitted into our hospital in recent 20 years were retrospectively analyzed. Results Twenty patients were in grade 3, 10 in grade 4,and 2 in grade 5. Operations were performed on all 32 patients. Twenty-five recovered, and 7 died. The mortality was 21.9%.Before the operation, peritoneal lavage fluid amylase rose in 10 patients(50%),and serum amylase rose in 12 patients(60%). Imaging examination included ultrasonography in 10 (diagnosed in 3),and CT in 8 (diagnosed in 3). Diagnosis was established only in 10 patients before the operation. Complications included fistula, abscess and pseudocyst. Conclusions Incidence of associated trauma is high. The preoperative diagnosis of pancreatic trauma is difficult. The rates of morbidity and mortality are high. Early diagnosis, careful exploration and proper surgery are very important. Intraoperative pancreatography or ultrasonographic scanning is useful in diagnosis. Somatostatin and human growth hormone are useful in the treatment.
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AIM: To investigate the effect of propofol on expression of protein kinase C (PKC) mRNA during pulmonary ischemia and reperfusion injury (PIRI) in rabbits. METHODS: Single lung ischemia and reperfusion animal model was used in vivo. The rabbits were randomly divided into three groups ( n =9 in each): sham operated group (sham), PIR group (I-R) and PIR+ propofol group (PPF). Changes of several parameters including malondialdehyde (MDA), superoxide dismutase (SOD), wet to dry ratio of lung tissue weight (W/D) and index of quantitative assessment of histologic lung injury (IQA) were measured at 60 minutes after reperfusion in lung tissue. Meanwhile the location and expression of PKC mRNA were observed. Lung tissue was also prepared for light microscopic and electron microscopic observation at 60 minutes after reperfusion. RESULTS: As compared with group I-R, PKC mRNA strongly expressed in intima and extima of small pulmonary artery as well as thin-wall vessels (mostly small pulmonary veins) in PPF group. The average optical density values of PKC-?、? and ? mRNA in small pulmonary veins PPF in group showed significantly higher than that in I-R group (all P
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Objective:To study the effect of pig interleukin 2(IL 2) eukaryon expression plasmid on cellular immune responses of BALB/c mice immuned with pcDNA PRRSV ORF5 DNA vaccine.Methods:BALB/c mice were immunized with pcDNA PRRSV ORF5 DNA vaccine and pig interleukin 2(IL 2) eukaryon expression plasmid by the routes of co injection and DNA vaccine injection alone respectively, with PBS and pcDNA3 1(+) as controls. Fluoresecence Activated Cell Sorter(FACS),T lymphocyte proliferation test(MTT) were used to detect the number of CD4 +、CD8 + and the T lymphocyte proliferation in peripheral blood of mice vaccinated.Results:ConA response of T lymphocytes in blood was higher in experiment group than the control group ( P