A highly sensitive fluorescence spectroscopic method was established for the selective determination of estradiol, which took advantages of the excellent molecular recognition capability of aptamer and the energy transfer between the specific fluorescent groups. The effects of the pH value, buffer constituent and concentration, the concentration of DNA, the experimental temperature and response time on the detection of estradiol were studied. Under the optimal conditions (50 mmol/L BR buffer solution with pH value at 7. 4, 1. 0×10-7 mol/L for each DNA strand, incubation at 45 ℃, response time 19 min), the change of the fluorescence intensity (ΔI) versus the logarithm of the concentration of estradiol ( lgC) was linear over a concentration range from 1. 0×10-11 mol/L to 5. 0×10-9 mol/L with good linear correlation (r=0. 9953). The limit of detection (LOD) was found to be 6. 0×10-12 mol/L (S/N=3). This method was successfully applied to the detection of estradiol in human urine, with the recovery in the range of 94. 0%-103. 5%. This method showed good precision and accuracy.

Objective To construct and express fusion toxin human IL15 mutant/TNF? mutant (hIL15M/TNF?M) in order to develop a targeted toxin capable of eliminating abnormal cells expressing high levels of hIL15 receptors. Methods DNA fragments of truncated hIL15M and TNF?M coding gene were joined in frame and cloned into T7 expression vector pET16b. The expressed fusion protein was purified from E. coli . Cytotoxicity of hIL15M/TNF?M was measured by MTT assay. Results E. coli transformed with the recombinant plasmid expressed an extra band with the expected size of 30?10 3. MTT assay indicated that PHA activated T cells were 13 times more sensitive to the cytotoxic effect of hIL15M/TNF?M than to the resting T cells. Conclusion hIL15M/TNF?M has high toxic effect against cells expressing high levels of IL15 receptors, suggesting that hIL15M/TNF?M has potential therapeutic effect on T cell leukemia and other diseases related with improperly activated T cells.

Objective To determine the clinical value of laparoscopic right hepatectomy (LRH) carried out following a standardized technique.Methods The medical records of 15 patients who underwent LRH at the First Affiliated Hospital of Soochow University were retrospectively reviewed.The perioperative indicators which included the operation duration,blood loss,tumor diameter,hospitalization duration and postoperative complications were analyzed.Results There was no conversion to open surgery.The operation duration was (251.1 ± 73.3) min.The resected tumor diameter was (8.5 ± 4.2) cm,the blood loss was (550.8 ± 343.6) ml,and the hospitalization duration was (10.2 ± 3.7) days.There was one patient who developed postoperative bile leakage.There was no other complication and there was no perioperative death.Conclusions The standardized technique of LRH was easy and effective.This technique shortened the operation duration and improved patient safety.

Objective To investigate the relation between the expression of toll receptor-4 (TLR-4) and the increase of blood brain barrier (BBB) permeability in the rats with acute pancreatitis.Methods Male Sprague-Dawley rats were randomly divided into 8 groups:control group;acute edematous pancreatitis (AEP) 2 h,6 h groups;acute necrotizing pancreatitis (ANP) 2 h,6 h,12 h,24 h,48 h groups with 8 rats in each group.The BBB permeability,pathological score of pancreas,TLR-4 expression were determined and the relationships between them were analyzed.Results The BBB permeability in control group;acute edematous pancreatitis (AEP) 2 h,6 h groups;ANP 2 h,6 h,12 h,24 h,48 h groups were 1.55±0.29,1.64±0.17,1.69±0.24,1.89±0.12,2.66±0.32,2.91±0.29,2.89±0.69 and 1.84±0.07,respectively;the pathological scores of pancreas were 0,2.38±0.92,3.13±0.64,8.50±1.07,9.75±0.71,10.25±1.28,11.13±1.25 and 10.13±1.13,respectively;there was no significant difference between AEP groups and control group,while there was significant difference between AEP groups and ANP groups (P＜0.05 orP ＜0.01).BBB permeability was correlated with pancreatic injury ( r = 0.626,P ＜0.01).There was no TLR-4 mRNA and protein expression in the control and AEP group,while they were significantly expressed in ANP groups,and the expression were positively related with BBB permeability ( r =0.208,P = 0.027 ).Conclusions BBB permeability was present in the course of ANP.Activation of TLR-4 signal pathway may be involved in the BBB permeability increase caused by ANP.

Objective To investigate the incidence of pleural effusion in acute panereatitis(AP)and the relation between pleural effusion and severity of AP.Methods The medical records of 766 patients with AP were analyzed retrospectively.The incidence of pleural effusion was documented and its relationship with age of onset,sex,Ranson score,CTSI,serum albumin and hospital day was analyzed.Results In the 766 AP patients,there were 129(16.8%)patients had pleural effusion.In 171 SAP patients,there were 91 (15.3%)patients had pleural effusion.In the 595 MAP patients,there were 129(16.8%)patients had pleural effusion.The difference in the incidence of pleural effusion between MAP and SAP patients was statistically significant(P＜0.05).The serum amylase,albumin,Ranson score,CTSI,and hospital day in patients with pleural effusion were(795±1013)U/L,(36±7)g/L,1.12±1.15,4.02±1.16 and(23.4±23.4)d,respectively;while the corresponding values in patients without pleural effusion were(592±856)U/L,(38±6)g/L,0.85±0.98,3.15±16.60 and(17.3±16.6)d,respectively.The difference in Ranson score,serum amylase,CTSI between the two groups was statistically significantly(P＜0.05 or＜0.01＝,the serum level of albumin in patients with pleural effusion was significantly lower than that in patients without pleural effusion(P＜0.01),the hospital day in patients with pleural effusion was significantly longer than that in patients without pleural effusion(P＜0.01).Pleural effusion of both sides was present in 92 patients,while pleural effusion of right side was present in 11 patients,pleural effusion of left side was present in 26 patients;the site of pleural effusion was not associated with the severity of AP,however,patients with pleural effusion of left side tended to have longer hospital day(P＜0.05).Conclusions Patients with pleural effusion during AP were more likely to be complicated with severe conditions,and the presence of pleural effusion may be a good marker for severity evaluation.

Objective To study the mutation of papillomavirus type 16 (HPV16) E6 gene, and to compare with the result of other researchers. Methods DNA was extracted from cervical cancer tissue sam-pies, then HPV16 E6 genes were amplified and sequenced using the HPV16 E6 specific primers from cervi-cal cancer DNA samples. Results HPV16 E6 gene mutations at the position of nt178 were found in 18 case from 35 cervical cancer samples, the mutation rate was 51.43%, resulting in nucleotide change of Asp to Glu. E6 mutations are found at the position of nt442 in 4 cervical samples. E6 mutation was found in 1 to 2 case in other nucleotide. Conclusion High frequency mutations of E6 gene of HPV16 at the position of nt178, nt442 have been found in cervical cancer samples in Hubei, China. The role of E6 gene mutation in cervical cancer development should be studied further.

OBJECTIVE To understand the urinary tract infections after cesarean section,the distribution of pathogenic strains,and drug resistance analysis in order to provide clinical treatment. METHODS The routine urine cultures of the 2652 case were identified and drug resistance was analyzed by K-B method. RESULTS From 458 isolated bacteria,Gram-negative bacilli were main,pathogen accounting for 80.13%,Escherichia coli and Klebsiella,accounted for 71.62% and 4.8% respectively;Gram-positive bacteria accounted for 14.41%,and fungi accounted for 3.71%. CONCLUSIONS G-bacteria in the urinary tract infection are in the absolute superiority;rational use of antibiotics to effectively control the urinary tract infections and avoid drug-resistant strains produceing is especially critical.

Objective To assess the value of CT or PET-CT with fluorine-18-labeled fluorodeoxyglucose (FDG) for the diagnosis of pulmonary mucosa-associated lymphoid tissue (MALT) type lymphoma. Methods The CT or FDG PET-CT findings in 14 patients with pathologically proved pulmonary MALT lymphoma were retrospectively analyzed. Results Lung lesions were unilateral in 7 patients and bilateral in 7 patients. Lesions presented as a single mass in 3 patients, as a single consolidation in 3 patients, as a nodule in 1 patient, as multiple nodules in 1 patient, as multiple patchy consolidations in 4 patients, as a mass with multiple nodules and patchy consolidations in 1 patients, as diffuse interstitial change in 1 patients. Air bronchogram was found in 9 patients and CT angiogram sign in 5 patients. On PET-CT, lesions showed heterogeneous FDG uptake in 2 patients, maximum standard uptake value was higher than 2. 5. Conclusion Imaging characteristics of pulmonary MALT lymphoma are single or multiple nodules or consolidations with air bronchogram on CT, and heterogeneous high FDG uptake on PET-CT.

OBJECTIVEWe aims to evaluate the therapeutic effect of maxillary protraction on maxillary maldevelopment using Coben analysis to illustrate the advantages of Coben analysis in identifying pathogenic mechanisms and in designing treatment plans for Class III malocclusions.

METHODSA total of 120 patients were diagnosed to have skeletal Class I maloc- clusions with maxillary maldevelopment. These patients were selected as the subjects of the present research. Maxillary protraction was exerted to promote maxilla growth. Cephalometric analysis was conducted by using Coben analysis and angle analyses of Beijing Medical University to collect data before and after maxillary protraction.

RESULTSAccording to Coben analysis measurements, the height of facies cranii increased after maxillary protraction. The depth of lower face decreased, whereas that of mid-face increased and the Ptm-A value increased significantly in the mid-face (P < 0.001). In the angle ana- lysis of Beijing Medical University, SNA, ANB, U1/NA, U1/SN, MP/SN, and Y-axis angles increased significantly (P < 0.001), whereas SNB, U1/L1 (P < 0.001), and L1/MP (P < 0.05) decreased.

CONCLUSIONMaxillary protraction has a significant effect on growing skeletal Class III malocclusions withthe chief mechanism of Ptm-A value just diagnosed as minor by Coben analysis. Coben analysis is visual and clear in identifying pathogenic mechanisms of Class III malocclusions.

Objective To determine the lower limit of detection (LLOD) and cut off values of K-ras mutation detection by peptide nucleic acid (PNA) clamping-PCR.Methods The genomic DNA of pancreatic cancer cell lines (PANC1 and SW1990) with codonl2,13 mutation and the genomic DNA of placenta with K-ras wild type were mixed and diluted serially into samples with different mutation rate (0,0.1％,0.2％,0.4％,0.8％,1.6％,3.1％,6.25％,12.5％,25％,50％),PANC1 cells with 1％ mutation rate and SW1990 cells with 30％ mutation rate and 4 samples with the quantity of DNA was 50,20,5,1 ng and 50,10,5,1 ng was prepared.Codon 12,13 mutation of K-ras was determined by PNA-PCR,and the mutation Ct values,overall Ct values were collected,and the △Ct values (mutation Ct values-overall Ct values) were calculated,and the tests were repeated for 10 times.ROC curve was used to analyze the △Ct values and determine the best cut off values for K-ras mutation,and the positive diagnostic rate,LLOD was evaluated.Results The mutation Ct,△Ct values of codon 12 mutation of PANC1 and codon 13 mutation of SW1990 of all the different mutation rates were statistically significantly different (P ＜ 0.05) when compared with negative control group,but the overall Ct values were not statistically significantly different from that of negative control group.For detection of K-ras codon 12 mutation by ROC curve,the relevant area of ROC curve (AUC) was 0.926,the optimum cut off value of △CT was 11,the sensitivity and specificity were 84％ and 100％,respectively,and the LLOD was 0.4 ng.For detection of K-ras codon 13 mutation by ROC curve,the relevant AUC was 0.906,the optimum cut off value of △CT was 9.5,the sensitivity and specificity were 71％ and 100％,respectively,and the LLOD was 1.5 ng.The mutation detection results of fixed rate further confirmed the LLOD.Conclusions This study successfully defines LLOD and cut off value of PNA clamping-PCR/K-ras method in detection of K-ras 12 and 13 codon mutations.This method meets the requirement of clinical application.