Objective To discuss the clinical values of 18 F-FDG imaging in screening of the recipients with liver transplantation.Methods By using positron emission tomography, 16 case of hepatocellular carcinoma and 21 case of hepatic cirrhosis with discompensation were subjected to 18 F-FDG imaging. The obtained images were fused with CT images. According to the result of the abnormal 18 F-FDG uptake in 18 F-FDG imaging and image fusion with CT imaging, extrahepatic malignant tumor was judged. The initial routine examinations showed no extrahepatic malignant tumor in these 37 cases , including primary extrahepatic carcinoma and extrahepatic metastasis of liver carcinoma.Results Among the 21 case of hepatic cirrhosis with discompensation, there were 5 cases of extrahepatic primary carcinoma and metastasis. In 16 cases of primary hepatocellular carcinoma, there were 7 cases of extrahepatic metastasis.Conclusion 18 F-FDG positron emission tomography imaging can find the extrahepatic carcinomas which can not be discovered by other examinations, which can provide more information for screening of the recipients undergoing liver transplantation.

Objective The newly developing gene therapy method and dominant negative mutants were bein g used as new promising HBV therapy method, and a dominant negative mutant of HB V X g ene we have reported in our previous report has some effects both on HBV replica tion and expression in transient expression, but the effects were interfered by persistent secretion of HBV in HepG 2 2.2.15 cell line in the experiment. To mak e sure the effects of dominant negative mutant of HBx gene, we established a HBx DN stably expressing cell clone, and evaluated the effects of HBx dominant negat ive mutant on HBV gene expression. Methods The prev HBx-GFP dominant mutant and the plasmids pRev Xwt, pRev GFP which contain the wild type X gene or GFP gene then transfected into HepG 2 2.2.15 cells by liposome. The HBsAg, HBeAg by in media were as sayed by RIA and HBV-related RNA were assayed by Northern blot. Results The pRev HBx-GFP, GFP and wild type X constructs can be effectively expressed in HepG 2 2.2.15 cells. The stable expressed HBx -GFP can significantly reduce HBeAg, HBeAg in media and the HBV-related RNA in HepG 2 2.2.15 cells, but not for pRev Xwt and pRev GFP. Conclusions The dominant negative mutant pRev HBx-GFP can significantly inhibit the HBV gen e expression. It also suggested that X gene may be one promising target for HBV gene therapy.

Objective To study the development a clinical useful microarray which can detect 4 most commonly and well documented sites for detection of lamivudine-resistance mutants in HBV lamivudine therapy. Methods (1) The selected 16 oligonucleotide probes to screen 4 mutated sites in YMDD region are immoblized in a specific treatment slice by GMS 417 Arrayer. The target 308 bp nucleic acids segment of HBV were amplificated and labeled with Cy5-dCTP fluorescent dye by PCR. After hybridization of target DNA with microarray, the microchips were scanned with Gene TAC LS IV scanner and the data were obtained after processed in computer. (2) To evaluated the specialty and reproducibility of the microarray, the plasmid with Leu515Met, Met539Ile, Met539Val,V542I mutations were condstructed and serve as positive sample. Another 50 sera of lamivudine treated Hepatitis B patients for 6 to 12 months as well as 50 sera without lamivudine administration were assayed by this microarray to evaluated the rate and genetype in lamivudine related resistance mutation. Results The microarray can identify a single base change of selected lamivudine resistance-related mutation and multiple mutation detection by a single assay as well. The specialty are well in agreement with sequencing for 98%. The reproducibility rate of repeat examination is range from 96%-100%. Conclusion The microarray of lamivuine resistance related mutation can identify a single base change as multiple mutations well. And its hight reproducible results may be useful for clinical monitoring lamivudine resistance related HBV mutation.

Objective To develop a clinically useful assay for detecting the mutations of HBV pre-C/BCP based on the pyrosequencing and accuracy, reproducibility and reliability of this assay was evaluated. Methods The pyrosequencing primers for HBV pre-C/BCP mutation were designed through the cluster analysis among one hundred HBV gene sequences. After the amplification of the fragment of pre-C/BCP with the template of pre-C/BCP mutation plasmids, the pyrosequencing method for pre-C/BCP detection was initially set up with this standard sample. The accuracy, reliability and reproducibility of the pre-C/BCP pyrosequencing were confirmed through the pre-C/BCP plasmids as a standard sample when compared with Sanger/genechip sequencing method pre-C/BCP pyrosequencing assay was applied for detecting pre-C/BCP mutation types of 60 chnical serum samples in HBV patients. Results The pre-C/BCP mutation detection assay based on pyrosequencing has been established in our study. The coincidence rate between pyrosequencing and Sanger squencing was 100%. The coincidence rate between the result of pyrosequencing and of genechip method was 91.7%. The reproducibility of this assay was 97. 8%. It indicates the pre-C/BCP pyrosequencing is a high-accurate method with, good-reproducibility and high-reliability. And multi-site detection can be achieved by pyrosequencing one time. A rare mutation T1758C was also detected. Conclusion Pyrosequencing for pre-C/BCP mutations assay is high-throughout method for simultaneous detection of multi-site mutation.

OBJECTIVE: To explore the reasons of poor sensation recovery after anterolateral thigh flap (ALTF) transplantation and to improve the design and sensation reconstruction of ALTF. METHODS: Lower limbs from 21 adult cadavers were chosen, and the lateral femoral cutaneous nerves (LFCN) were dissected. Their courses, shape, distribution and anatomic variation were observed, the distance from original sites of LFCN's posterior and anterior branches to the anterior superior iliac spine (ASIS) were measured, and the external diameter of their main trunks,after piercing out from the inferior margin of inguinal ligament or sending out their posterior branches and their anterior branches, were measured. RESULTS: The plane which the posterior branches sent out from the LFCN located at 4.8 (0~16.9) cm below the ASIS. The plane which the anterior branches sent out from the LFCN located at 14.2 (6.7~24.1) cm below the ASIS. There were 6 branches of the LFCN, namely ordinary three branches (9/21), high-level posterior branch (5/21), posterior branch absent (3/21), anterior branch absent (1/21), tiny branch, (2/21) and LFCN absent (1/21). The section of the LFCN was oblate, the external diameter of the LFCN,s main trunk after piercing out from the inferior margin of inguinal ligament or sending out its posterior branch and its anterior branch was 2.68 (1.18-4.52) mm, 2.18 (0.80-4.10) mm and 1.63 (0.44-2.60) mm, respectively. CONCLUSION Poor sensation recovery after ALTF transplantation is due to anatomic variation of the posterior branch of the LFCN. The sensory recovery of ALTF may be improved if the 2/3 low to median part of the anterolateral thigh and the main trunk of the LFCN or its plane before sending out the anterior branch is chosen for anastomosis.
Cadaver ; Female ; Humans ; Ilium ; anatomy & histology ; Inguinal Canal ; anatomy & histology ; Male ; Skin ; innervation ; Surgical Flaps ; innervation ; Thigh ; innervation

Anti-HIV Agents ; pharmacology ; DNA Mutational Analysis ; methods ; DNA, Viral ; genetics ; Drug Resistance, Viral ; genetics ; Hepatitis B virus ; drug effects ; genetics ; Lamivudine ; pharmacology ; Mutation

Objective To evaluate the image quality of coronary CT angiography (CTA) and the diagnostic accuracy for the assessment of coronary artery stenosis with a 64-slice CT scanner.Methods 120 patients with suspected coronary artery disease were studied by ECG-gated 64-slice spiral CT (120 kV, 850 mAs,0.33 s-rotation time,0.6 mm collimation);60 of them also underwent conventional coronary angiography(CCA).Patients with heart rates above 70 bpm received oral?-blockade before the scan. Enhanced scan was performed with an intravenous injection of 60-70 ml contrast agent (370 mg l/ml) and 50 ml saline chaser bolus at 4 ml/s.Images were retrospectively reconstructed under different reformations multiplanar reconstruse-tion(MPR),maximum intensity projection(MIP)and volume rendering technology (VRT)and reviewed by two observers.Image quality was assessed using a 3-point grading scale(excellent, sufficient and non-diagnostic)and the visibility of coronary branches was evaluated.The left main,left anterior descending,left circumflex and right coronary arteries were screened for the presence of over 50% stenosis.Results Sixty-nine percent of the included patients received?-bloekade.The average heart rate was 61?8 bpm and the scan time was 11.9?1.3 s.In the evaluation of image quality with VRT images, 77% patients′images were excellent,18% sufficient,and 5% non-diagnostic.All of the first,most of the second and third,and parts of the fourth subsegment branches could be shown with MPR,MIP and VRT reconstruction images.MPR and MIP displayed better visualization than VRT in showing small branches. Compared with CCA,the sensitivity,specificity,positive and negative predictive value of CTA to identify over 50% stenosis were 93%,98%,87% and 99%,respectively.Conclusion 64-slice spiral CT coronary angiography is a noninvasive and fast method that allows reliable diagnosis of coronary artery stenosis with high image quality.