Objective To quantitatively analyze the K-ras gene mutation at codon 12 in pancreatic cancer tissues and the relationship between K-ras gene mutation and clinicopathological parameters. Methods Quantitative detection of K-ras gene at codon 12 in 93 pairs of pancreatic cancer and adjacent tissues were performed by using PNA-mediated PCR clamping with two different fluorescence labeled probes. The quantity of mutation was expressed by percentage of mutation. The percentage of K-ras gene mutation = the copy of K-ras mutation/(copy of wild type K-ras + copy of K-ras mutation) × 100%. Results The percentage of mutation of K-ras gene at codon 12 in pancreatic cancer and adjacent tissues were 83.9% and 65.6%, and the difference was statistically significant(P ＜ 0. 05); and the quantity of mutation were (13.385 ± 1. 745) % and (2. 246 ±0. 728) %, and the difference was also statistically significant(P ＜ 0. 05). The quantity of mutation of K-ras gene at codon 12 was not associated with clinicopathological parameters. Conclusions The percentage of K-ras gene mutation, as well as the quantity of K-ras gene mutation was different in pancreatic carcinoma and adjacent tissues.

Objective To investigate functional role and clinical significance of the methylation in the promoter of TPAP gene in the development and progression of pancreatic cancer.Methods Surgically resected specimen from 68 patients who were pathologically diagnosed as pancreatic cancer in Changhai Hospital from July 2006 to August 2009 were collected.The methylation in the promoter of TPAP gene in tumor and nontumor adjacent tissue was detected by methylation specific PCR.Results The methylation rate of tumor and non-tumor adjacent tissue was (0.214 ± 0.057) ％ and (0.084 ± 0.096) ％,respectively,and pancreatic cancer tissue had significantly higher methylation rate than the adjacent tissue.Hypermethylation of TPAP gene was not correlated with age,gender,tumor differentiation,lymphatic metastasis,serum CEA and CA19-9,but was positively correlated with distant metastasis.Conclusions Hypermethylation in the promoter of TPAP gene may participate in the invasion and metastasis of pancreatic cancer and the hypermethylation of the promoter is closely associated with the tumorigenesis and development of pancreatic cancer.

ObjectiveTo investigate the clinical significance of quantitative detection of K-ras codon 12 and 13 mutations in the tissues of pancreatic cancer and related pancreatic diseaaes. Methods One hundred and thirty samples from surgically removed pancreatic tissue with a conclusive pathological diagnosis (105 cases of pancreatic ductal adenocarcinoma,8 cases of pancreatic adenosquamous carcinoma of the pancreas,2 cases of pancreatic mucinous adenocarcinoma,3 cases of pancreatic endocrine carcinoma,6 cases of duodenal and papillary adenocarcinoma and 6 cases of benign pancreatic diseases ) were collected.Quantitative detection of K-ras codon 12 and 13 mutations was performed by the method of peptide nucleic acidmediated PCR clamping with two different fluorescence labeled probes.Mutation number ＞ 100 copies was used as the criteria to calculate the positive mutation rate.ResultsThe median and quartile of K-ras codon 12mutations of pancreatic ductal adenocarcinoma,adenosquamous carcinoma of the pancreas,pancreatic mucinous adenocarcinoma,pancreatic endocrine carcinoma,duodenal and papillary adenocarcinoma and benign pancreatic diseases were 4062 (495,10800),238 (45,8420),15 (9,21),3 (3,16),2283 (73,5037)and 21(8,56),and the positive mutation rates were 84.8％ (89/105),50.0％ (4/8),0,0,66.7％ (4/6)and 16.7％ (1/6).The quantity of K-ras codon 12 mutation in pancreatic ductal adenocarcinoma was not statistically different from those of adenosquamous carcinoma,duodenal and papillary adenocarcinoma,but it was significantly higher than those in pancreatic mucinous adenocarcinoma,pancreatic endocrine carcinoma,and benign pancreatic diseases (P ＜0.05).The area under ROC of K-ras codon 12 mutation in pancreatic ductal adenocarcinoma was 0.727.The sensitivity and specificity of the K-ras codon 12 mutation for the diagnosis of pancreatic ductal adenocarcinoma were 84.8％,64.0％,respectively.The quantity of K-ras codon 12 was associated with survival of patients with pancreatic ductal adenocarcinoma.The quantity of K-ras codon 13 mutations and the positive mutation rates in pancreatic ductal adenocarcinoma was not statistically different from other pancreatic diseases.ConclusionsThe quantity of K-ras codon 12 mutation has good differential diagnostic and prognostic prediction value for pancreatic ductal adenocarcinoma.

Objective To compare the effect of conventional and hyperoxia management strategy during deep hypothermia in patients with DeBake type 1 aortic dissection or aortic arch aneurysm undergoing total aortic arch replacement.Methods 32 adult patients undergoing total aortic arch replacement were randomly allocated to one of two groups(n=16 each):conventional(C)and hyperoxia group(H).The patients had no history of cerebral vascular disease.Left radial artery and dorsal artery of left foot were cannulated for monitoring of blood pressure of upper and lower limbs.Right internal jugular vein was cannulated for CVP monitoring and administration of drug and fluid.Anesthesia was induced with etomidate 10-15 mg,fentanyl 5-10 ?g?kg~(-1) and pancuronium 0.1 mg?kg~(-1) and maintained with fentanyl(total amount was＜20 ?g?kg~(-1)),isoflurane and pancuronium after tracheal intubation.Intermittent i.v.boluses of diazepam,sodium thiopental or propofol were given during cardiopulmonary bypass(CPB).Another catheter was inserted into right internal jugular vein eephalad until resistance was met.The tip of the catheter was at the level of mastoid process.The hyperoxia management involved the following steps:FiO_2 was gradually reduced with decreasing body temperature(T_0)from 70%(36～ 37℃)to 60%-40%(35.9-34℃),38%-30%(32-26℃),30%(26-24℃)and finally to 21%.When nasopharyngeal T_0 was reduced to 22℃ or 5-10 min before selective cerebral peffusion(SCP),FiO_2 was raised to 60%-100% to maintain PjvO_2＞20 mm Hg or SjvO_2＞60%.FiO_2 was maintained at 60%-100% during SCP until T_0 was rewarmed to 22℃,then reduced to 30%.FiO_2 was then gradually increased to 40%(when T_0 reached 28℃),to 50%-70% (34-37℃)and finally to 80%(T_0＞37℃).Blood samples were taken from jugular venous bulb and arterial port of oxygenator for determination of PjvO_2,SjvO_2 and PaO_2 before skin incision (T_1),at 15 min of CPB(T_2),10 min of SCP(T_3),5 min after descending aorta unclamping(T_4),5 min after left subclavian artery unclamping(T_5),5 min after left common carotid artery unclamping(T_6),anonymous artery unclamping(T_7),when nasopharyngeal To returned to 35℉(T_8)and 10 min after CPB was terminated(T_9).The awakening time and the duration of ICU stay(days)were recorded.Pre- and postoperative neurological examination and brain CT scan were performed.Results All patients survived the operation and were discharged from hospital.No new brain infarction occurred.Transient neurologic dysfunction occurred in 2 patients in group H and 3 patients in group C.There was a positive linear relationship between PaO_2 and PjvO_2 during deep hypothermia in group H (r=0.541,P＜0.01).The PjvO_2 and SjvO_2 were significantly higher in group H than in group C.The awakening time and the ICU stay were significantly shorter in group H than in group C.Conclusion The hyperoxia management strategy can provide clinical prognosis than the conventional management strategy during deep hypothermia for total aortic arch replacement by supplying more dissolved oxygen.

0.05).Among the 26 pregnancies,there occurred one ectopic pregnancy and four cases of spontaneous abortion.In the 13 delivery cases,there were one premature delivery,two cases of premature rupture of the membrane,and ten cases of cesarean section.The sample height of the cone was less than 2.0 cm in the nine delivery cases,and the mean width of the cone was over 2.5 cm.Conclusion No evidence of secondary infertility caused by cervical conization was found.There was also no significant increase in the number of either premature delivery cases or low birth weight infants. The sample height of the cone might play a more important role in the pregnancy outcome than the width, which still needs to be further verified by larger studies.

Objective To detect the microRNAs in fecal with patients of pancreatic cancer, and evaluate its diagnostic value. Methods Stool samples were collected from three group persons including 29 pancreatic cancer, 22 chronic pancreatitis and 13 normal controls. The total fecal microRNAs were extracted.The quantity of miR-16, miR-21, miR-155, miR-181a, miR-181b, miR-196a, and miR-210 were detected by using real-time PCR, and miR-16 was used as reference gene. ROC AUC was used to evaluate the diagnostic value for pancreatic cancer. Results MicroRNAs were efficiently obtained from stools, and independent experiments showed high reproducibility for microRNAs extraction and detection. The expression of miR-181b,miR-196a, miR-210 in fecal was 2.22 ±0.64,2.78 ±0.14, 5.55 ±0.38 in pancreatic cancer; 1.42 ±0.39,3.88 ± 0.85,5.39 ± 0.69 in chronic pancreatitis; 0.32 ± 0.40, 1.14 ± 0.98,4.23 ± 0. 99 in normal controls;the three microRNA expressions in pancreatic cancer were group and CP group significantly higher than those in normal controls ( P ＜ 0.05 ). But there was no significant difference between pancreatic cancer group and chronic pancreatitis group. AUC of pancreatic cancer / normal controls miR 18lb was 0.745(95% CI 0. 597-0.894), the sentivity, specificity for pancreatic cancer was 84.6% and 51.7%. AUC of miR-210 was 0. 772(95% CI0.629-0.914), the sentivity, specificity for pancreatic cancer was 84.6% and 65.5%, and the difference was statistically significant (P ＜0.05). miR-196a was no significant for the diagnosis of pancreatic cancer, but the expression of miR-196a was correlated with the tumor size (r = 0.516, P = 0.041 ).Conclusions The extraction and detection of the fecal microRNAs were non-invasive and reproducible. The expression of miR-181b and miR-210 was increased in stool of patients with pancreatic cancer, and may be potential biomarker for pancreatic cancer.

nd hypermethylation of HHIP was detected in pancreatic juice,which may be a useful marker in the diagnosis of PCa.

Objective To investigate the RADIL mRNA expression in pancreatic carcinoma and to evaluate its clinical significance.Methods Fluoesecent quantitative PCR (FQ-PCR) was used to detect the RADIL mRNA expression in 40 patients with pancreatic carcinoma and adjacent tissue and in 5 healthy adult with normal pancreatic tissue and to observe its relationship with clinicopathologic parameters.Results RADIL mRNA was expressed in pancreatic carcinoma and adjacent tissue, as well as normal pancreatic tissue, and the relative expression was 2.263 ± 3.826, 5.425 ± 8.858 and 8.559 ± 4.214, respectively.There was statistically significant difference among the three groups (P ＜0.05 ).RADIL mRNA expression was closely related with the metastasis and differentiation grade ( r = -0.312 and -0.294, P ＜ 0.05 ), however, it was not significantly related to tumor site, tumor size, CA19-9, TNM staging, sex and age.Conclusions RADIL gene may have an inhibitory effect on the pancreatic cancer.

Objective To investigate the diagnostic value of the K-ras mutations in FNA samples for early detection of pancreatic cancer. Methods FNA samples of 27 patients with pancreatic cancers, 9 patients with other malignant tumors and 14 patients with non malignant pancreatic mass (NMPM) were collected. DNA was extracted, and K-ras gene was amplified through PNA-mediated PGR clamping, the products were sequenced to determine the mutation type. Results The positive rate of K-ras mutations in pancreatic cancers,other malignant tumors and NMPM were 88.9%, 44.4%, 35.7%. There was significant difference in K-ras gene mutations in FNA samples between pancreatic cancer and other malignant tumors ( P = 0. 013 ) and NMPM ( P = 0. 001 ). The sensitivity, specificity, positive predictive value, negative predictive value,accuracy of K-ras mutations in FNA samples of pancreatic cancers were 88.9%, 55.6%, 85.7%, 62.5%,80.6% when compared with other malignant tumors, and the difference between the two groups was significant (P =0. 013) ;Those were 88.9%, 64.3%, 82.8%, 75.0%, 80. 5% when compared with NMPM, and the difference between the two groups was significant ( P = 0. 001 ). When cytology of FNA samples and K-ras mutations was combined, the positive rate of pancreatic cancer was up to 96.3%. Conclusions The detection of K-ras mutations in EUS-FNA samples helped improve the positive diagnostic rate of pancreatic cancer.

Objective To investigate the methylation status of PCDH8 gene in pancreatic carcinoma.Methods Methylation of PCDH8 gene in 2 samples of normal pancreatic tissues and 6 pancreatic carcinoma cell lines (PANC1, ASPC1, BxPC3, CFPAC, PaTu8988 and SW1990) was detected by the methylationspecific PCR (MSP) method. The expression of PCDH8 mRNA was detected with 5-Aza-2-deoxycytidine (5-Aza-dC) treatment, a kind of DNA methyltransferase (DNMT) inhibitor in 6 pancreatic carcinoma cell lines by real-time-PCR. Results The methylation of PCDH8 gene was not detected in normal tissues, while it was partially methylated in PANC1, BxPC3, CFPAC and it was totally methylated in PaTu8988, ASPC1, SW1990.PCDH8 mRNA was expressed in PANC1, SW1990, PaTu8988 and the relative quantities of mRNA expression (RQ) were 1.576 ± 0.648, 0.013 ± 0.008, 0.002 ± 0.001; PCDH8 mRNA was not expressed in BxPC3,CFPAC, ASPC1. After 5-Aza-dC treatment, PCDH8 mRNA was expressed in PANC1, ASPC1, BxPC3,CFPAC, PaTu8988, SW1990 and the relative quantities of mRNA expression all significantly increased, and they were 7. 463 ± 2.628, 10. 696 ± 1.539, 7.852 ± 2.762,421.815 ± 1.493, 118.595 ± 4.089, 6.690 ±1.884. Conclusions The methylation of PCDH8 gene may be the major mechanism of down-regulated expression of PCDH8 gene in pancreatic carcinoma.