Objective To establish a method for detecting K-rag gene point mutation in pancreatic adenocarcinoma based on the pyrosequencing and to compare its performance with that of Sanger sequencing.Methods Genomic DNA was extracted from formalin-fixed,paraffin-embedded pancreatic tissues including 49 pancreatic adenocarcinoma,10 normal pancreas,11 chronic pancreatitis,18 benign pancreatic tumor,7 insulin carcinoma,9 ampullary carcinoma,7 bile duct carcinoma and 7 duodenum papillary adenocarcinoma tissues.K-rag gene point mutations at codon 12 were analyzed by pyrosequencing and Sanger sequencing,respectively.Results No mutant K-ras gene Wag detected in normal pancreas,chronic pancreatitis,benign pancreatic tumor,insulin carcinoma,ampullary carcinoma,bile duct carcinoma and duodenum papillary adenocarcinoma tissues by either pyrosequencing or Sanger sequencing.K-rag gene point mutation was detection rate in pancreatic adenocarcinoma tissues was 71.4%(35/49)by pyrosequencing and 61.2%(30/49)by Sanger sequencing,respectively.Conclusions Pyrosequencing is more sensitive than Sanger sequencing and is also accurate,rapid and of high throushput in detecting mutant K-ras gene.It may serve as a practical method for fast batch analysis of clinical samples.

Objective To investigate the safety of thalidomide in the treatment of immune-related bowel diseases for providing clinical reference.Methods Thirty-five patients with immune-related bowel diseases (31 Crohn's disease,2 ulcerative colitis and 2 Behcet's disease) treated with thalidomide were enrolled in this study.The incidence,type,severity,duration of thalidomide related adverse drug reaction (ADR) and the dose-effect relationship of neurotoxicity were analyzed.Results All the patients were treated with a mean dose of thalidomide (109.29 ± 30.37) mg/d for (18.8 ± 12.4) months,and 33 occurred ADR.The three most frequent ADR were numbness [51.4％ (18/35)],somnolence [48.6％ (17/35)] and dermatitis [37.1％ (13/35)].The median time to development of these three ADR were 6.50,0.25,and 1.00 months,respectively.Severe ADR leading to withdrawal accounted for 20.0％ (7/35),including reasons of peripheral neuritis (3/7),dermatitis (2/7) and myelosuppression (2/7).The incidence of peripheral neuritis was not significantly related to the maximal and initial dose of thalidomide (P ＞ 0.05).Conclusions Although the incidence of ADR was relatively high during the treatment of thalidomide,most of them were mild and well tolerated.Thalidomide can be safely used in patients with immune-related bowel diseases under close monitoring.

Objective:To investigate the value of MRI and MSCT in TNM staging of laryngocarcinoma.Method:Thirty-seven patients with laryngocarcinoma were underwent by contrast enhanced scan and muhiplanar reconstruction.Thirty-five patients with laryngocarcinoma were underwent contrast enhanced muhislice spiral CT,which of them were done by MPR.There are 28 cases which were scan by MRI and MSCT in the two former nd we contrasted the accuracy rating in laryngeal manifestation of abnormality.In the former two groups,we observed them the variability in the aspect of lymphaden metabasis and TNM staging.Result:In all 28 cases,the MRI had better accuracy rating in displaying the parts of preepiglottic space,larynx side interspace,lingual root,eck tissue,vocal cord.In the TNM staging,there was no difference in stage one in accuracy rating,as the stag stepping up,the accuracy rating of MRI had became better.The last result was that the two methods had difference in staging.In the two methods,MSCT had better sensitivity,specificity and accuracy rating.Conclusion:RI and MSCT had good accuracy rating in TNM staging,MRI has better accuracy rating in some of laryngeal,but as the lymphaden metabasis,the MSCT was better.There were variability in staging,and the MRI was better.

Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; CD2 Antigens ; metabolism ; Child, Preschool ; Chromosome Breakage ; Cyclophosphamide ; therapeutic use ; Doxorubicin ; therapeutic use ; Female ; Follow-Up Studies ; Humans ; Ki-1 Antigen ; metabolism ; Lymphoma, Large-Cell, Anaplastic ; drug therapy ; metabolism ; pathology ; Mucin-1 ; metabolism ; Prednisone ; therapeutic use ; Receptor Protein-Tyrosine Kinases ; genetics ; metabolism ; Vincristine ; therapeutic use

Adult ; CD56 Antigen ; metabolism ; Diagnosis, Differential ; Epstein-Barr Virus Infections ; Herpesvirus 4, Human ; isolation & purification ; Humans ; Killer Cells, Natural ; pathology ; Lymph Nodes ; pathology ; Lymphoma ; metabolism ; pathology ; virology ; Lymphoma, Extranodal NK-T-Cell ; pathology ; Male

Objective To explore a method of establishing a canine model of intracerebral hemorrhage (ICH) by puncturing the main branch of middle cerebral artery (MCA) under ultrasonic guidance and proceed a pilot study of contrast-enhanced ultrasound (CEUS) of ICH.Methods Twelve adult healthy mongrel dogs were enrolled in the experiment.In the condition of bone window,the main branch of MCA was punctured to bleed under the ultrasonic guidance.CEUS of brain was performed every 30 minutes until hematoma formed and another CEUS was performed at 24 hours.Then brains were collected for pathologic examination after dogs were euthanized.Results Eleven of twelve dogs developed ICH successfully that located in the Out edge of basal ganglia,which was confirmed by CEUS and autopsy.One dog died within 6 hours due to intraventricular hemorrhage which was confirmed by autopsy.The hematoma diameters were measured as (22.4 ± 7.1)mm by CEUS before euthanasia and as (21.6 ± 6.9)mm on autopsy.There was no significant difference between the two methods of measurement (t =1.521,P =0.1565).The characteristics of active bleeding and hematoma on ICH were performed in the CEUS imaging.Conclusions A model of dog ICH by puncturing the main branch of MCA under the guidance of ultrasound was established successfully,and it proved to be simple,effective and repeatable.The imaging characteristics of this model are in good accordance with those of ICH in patients.

Objective To conclude the characterization of renal cellular carcinoma(RCC)with contrast enhanced ultrasound.Methods Seventy patients(seventy-two nodules)with RCC,which were confirmed by operation and biopsy underwent conventional ultrasound and contrast-enhanced ultrasound(CEUS).Microbubble agents SonoVue and contrast pulse sequence(CPS)were used.The conventional uItrasonographic characterization and the enhancement patterns of lesions were analyzed.Results On baseline sonography,the numbers of lesions that showed hypoechogenicity,isoechogenicity,and hyperechogenicity were 44.4%(32/72),25.0%(18/72)and 30.6%(22/72),respectively.Only 28 lesions(38.9%)showed flow signals on color Doppler sonography,the mean maximum velocity of which WSS(43.7±16.8)cm/s(range,24.8-95 cm/s),and the mean resistance index was 0.635±0.11(range.0.52-0.83).Sixty-three(87.5%)lesions were hyper-vascular in cortical phase.Among them forty-eight(76.2%)lesions were hypo-enhanced,and fifteen(23.8%)lesions were still hyper-vascular in late phase.The remaining nine hypervascular nodules in cortical phase were still hyper-enhancing in late phase.Fifty-four(75.0%)lesions were inhomogeneous enhancement.and pseudocapsule was observed in sixty-three(87.5%)RCC lesions.Conclusions The enhancement patterns of RCC are characteristic,and CEUS may be helpful in differential diagnosis of focal renal lesions.

Objective To investigate the usefulness of parametric imaging of contrast-enhanced ultrasound(CEUS) in imaging hepatocellular carcinoma (HCC) using dynamic vascular patterns (DVP). Methods Thirty clinically or pathologically proven HCCs that had undergone CEUS were randomly included. SonoLiver CAP sofeware was used to analyze the CEUS images and reconstruct DVP parametric images. Results The rise time, time to peak and mean transit time were (16. 72±11. 07) s, (29. 92±14. 13) s,(115. 03±90. 91)s in HCC versus (26. 59±9. 60) s, (41.67±12. 59) s, (159.26±123. 74) s in the surrounding liver parenchyma (all P <0. 05). The perfusion index was (90. 41±102. 49) % in HCC versus (54. 10±24. 99)% in surrounding liver parenchyma( P = 0.044). DVP curve and DVP parametric image could both be divided into three types:washout,non-washout and cystic type. The percentages of which were 76.7% (23/30), 20.0% (6/30) and 3.3% (1/30) in DVP curves, respectively, and 66.7% (20/30), 30.0% (9/30) and 3.3% (1/30) in DVP parametric images,respectively. Conclusions Parametric image of CEUS could demonstrates the difference of flow perfusion static between HCC and surrounding liver parenchyma dynamically and directly.

OBJECTIVE: To investigate the value of MRI and MSCT in TNM staging of laryngocarcinoma. METHOD: Thirty-seven patients with laryngocarcinoma were underwent by contrast enhanced scan and multiplanar reconstruction. Thirty-five patients with laryngocarcinoma were underwent contrast enhanced multislice spiral CT, which of them were done by MPR. There are 28 cases which were scan by MRI and MSCT in the two former and we contrasted the accuracy rating in laryngeal manifestation of abnormality. In the former two groups, we observed them the variability in the aspect of lymphaden metastasis and TNM staging. RESULT: In all 28 cases, the MRI had better accuracy rating in displaying the parts of preepiglottic space, larynx side interspace, lingual root, neck tissue, vocal cord. In the TNM staging, there was no difference in stage one in accuracy rating, as the stag stepping up, the accuracy rating of MRI had became better. The last result was that the two methods had difference in staging. In the two methods, MSCT had better sensitivity, specificity and accuracy rating. CONCLUSION MRI and MSCT had good accuracy rating in TNM staging, MRI has better accuracy rating in some of laryngeal, but as the lymphaden metastasis, the MSCT was better. There were variability in staging, and the MRI was better.
Adult ; Carcinoma, Squamous Cell ; diagnostic imaging ; pathology ; Female ; Humans ; Laryngeal Neoplasms ; diagnostic imaging ; pathology ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Neoplasm Staging ; Tomography, Spiral Computed

OBJECTIVETo investigate the effect of 5-Aza-2'-deoxycytidine (5-Aza-dC) on TIP30 gene expression and the relationship between TIP30 expression and the sensitivity to 5-fluouracil (5-Fu) in colorectal cancer cells.

METHODSThe methylation profile of TIP30 gene in HCT116 colorectal cancer cells was determined by methylation-specific PCR. The levels of TIP30 mRNA and protein were determined by RT-PCR and Western blot after the 5-Aza-dC treatment. MTT assay was used to detect the chemosensitivity of HCT116 cells to 5-Fu.

RESULTSTIP30 gene displayed complete DNA methylation in the HCT116 cells without 5-Aza-dC pretreatment. After the 5-Aza-dC treatment for 3 days, only demethylating PCR amplification product was detected and TIP30 gene showed DNA demethylation. With the prolongation of the time of removal of 5-Aza-dC treatment, methylated and demethylated PCR amplification products were observed and TIP30 gene displayed both DNA methylation and DNA demethylation in the colorectal cancer cells. At the day 10 after removal of 5-Aza-dC, methylating PCR amplification product appeared and TIP30 gene showed DNA methylation. No expressions of TIP30 mRNA and protein were detected in the HCT116 cells untreated with 5-Aza-dC. After the treatment of 5-Aza-dC for 3 d and then removed the 5-Aza-dC, the expressions of TIP30 mRNA and protein were increased obviously. With the prolonged time after 5-Aza-dC removal, the expressions of TIP30 mRNA and protein decreased and reached the lowest level on day 10. The IC50 values of 5-Fu were 41.62, 33.17 and 4.96 µg/ml in the HCT116 cells pretreated with 5-Aza-dC, d0 and d10 with the drug removal after drug treatment for 3 d, respectively.

CONCLUSIONSThe results of this study show that the expression of TIP30 gene may be associated with its DNA methylation status and may affect the sensitivity of colorectal cancer cells to 5-Fu.

Acetyltransferases ; genetics ; metabolism ; Antimetabolites, Antineoplastic ; pharmacology ; Azacitidine ; analogs & derivatives ; pharmacology ; Cell Proliferation ; drug effects ; CpG Islands ; genetics ; DNA Methylation ; drug effects ; Drug Resistance, Neoplasm ; drug effects ; Fluorouracil ; pharmacology ; Gene Expression Regulation, Neoplastic ; HCT116 Cells ; Humans ; Inhibitory Concentration 50 ; RNA, Messenger ; metabolism ; Transcription Factors ; genetics ; metabolism