Objective To detect nucleos(t)ide-resistance mutations in hepatitis B virus (HBV) covalently closed circular DNA (cccDNA) isolated from hepatocytes of patients with chronic HBV infection and to analyze the correlation between the mutations found in cccDNA and relaxed circular DNA (rcDNA). MethodsForty patients with chronic HBV infection were investigated. Preoperation serum samples and non-tumor liver tissues were collected.Intrahepatic HBV cccDNA and rcDNA were selectively extracted by co-precipitation of sodium dodecyl sulphate-protein and QIAamp DNA Mini Kit, and further purification with plasmid-safe ATP-dependent DNase (PSAD).Thereafter,cccDNA were amplified by selective polymerase chain reaction (PCR) or nested PCR using the primers spanning both the two gaps in HBV genome and covering the common mutations associated with nucleoside analogues resistance (rt169- rt250).Intrahepatic HBV rcDNA and pre-operation serum HBV rcDNA were also extracted and then amplified by PCR.The PCR products were then purified and sequenced.Results Among the 40 patients,intrahepatic HBV cccDNA were detected in 31 patients,and HBV rcDNA were detected in liver samples of 35 patients and pre-operation serum samples of 21 patients. The PCR products amplified from these samples were all successfully sequenced.rtM204Ⅰ mutation was detected in intracellular HBV cccDNA,rcDNA and serum HBV rcDNA in 2 patients.Both rtM204Ⅰ and rtQ215H were detected in intrahepatic HBV cccDNA and rcDNA in 2 patients,while no corresponding mutation was observed in serum HBV rcDNA of these two patients.Three variants including rtM204V,rtM204V and rtV173L-rtL180M-rtM204V were detected in serum HBV rcDNA in 3 patients,while corresponding mutants were not detected in intracellular HBV cccDNA or rcDNA of these patients.Condsions The results suggest that antiviral nucleos(t) ide resistance mutations can be found in HBV cccDNA in chronic hepatitis B patients. The dominant resistant mutation found in intrahepatic HBV cccDNA/rcDNA may be different from that in serum HBV rcDNA.

Objective To investigate the morphological changes of whole brain gray matter in patients with postherpetic neuralgia(PHN),and evaluate the correlation between morphological changes of whole brain gray matter,visual analogue scale (VAS) and duration of PHN.Methods By using VBM-DARTEL method,firstly the 3D T1WI structure images of 17 patients with PHN and 17 normal control were preprocessed,then the segmented gray matter volume was compared between the two groups, and the correlation between difference of cerebral gray matter volume,VAS and duration of PHN were analyzed.Results Compared with the normal control, gray matter decrease in PHN group was found in bilateral orbital frontal regions(orbital gyrus,rectus gyri),left inferior frontal gyrus,left insular lobe, left caudate, right cingulate gyrus,left superior temporal gyrus, left cuneus, bilateral cerebellum posterior lobe and bilateral cerebellum anterior lobe.And gray matter increase in PHN group was found in bilateral superior frontal gyri, left middle frontal gyrus, vermis, pons.The gyrus volume change for two hemispheres was asymmetric (the left side greater than the right side).These different brain region changes in gray matter volume for patients with PHN were not correlated with VAS and duration of PHN.