Objective To analyze the mutation of PreS-S region in occult hepatitis B virus(OHBV) in HBV infected persons with positive HBsAb and investigate the biological mechanisms of the special infectious model.Methods A total of 38 HB-sAb positive OBI serum samples were amplified by Nested PCR and sequenced,HBV genotype and serotype were determined.The amino acid sequences of OHBV were compared to the corresponding sequence of wild-type strains of similar genotype obtained from the GenBank database.Results PreS-S segment of 11 samples were obtained and 8 samples were sequenced successfully.Among which,5 were genotype C and 3 were genotype B.Genotype B were all serotype adw,while genotype C were 1 adw and 4 adr.The mutation rates of PreS-S region,the immunoreactive area and the major hydrophilic region (MHR) were higher in OHBV than the wild-type strains (2.6％ vs 0.8％,x2 =40.23,3.2％ vs 0.3％,x2 =52.13,3.6％ vs 0.6％,x2 =13.25,all P＜0.01) and the substitutions of I126T,Q129R,M133T,F134I,D144E,G145K in α determinant were found in OBI samples.The mutation rate of amino acids in PreS-S region was higher in genotype C than genotype B (3.5％ vs 1.2％,x2--15.98,P＜0.01),meanwhile,the mutation rates in MHR,α determinant and immunoreactive region were higher in genotype C too,but no statistical significance was attained (4.7％ vs 1.7 ％,x2 =2.96,3.6 ％ vs 2.9％,x2 =0.25,4.1％ vs 2.3％,x2 =3.59,all P ＞0.05).Conclusion Mutations in PreS-S region,especially in immunoepitope,might change the virus'immunogenicity leading to escape from immune response and cause OBI with HBsAb positive.

BACKGROUNDOccluders licensed for clinical use are not fit for some special Krichenko E patent ductus arterioses. The Amplatzer vascular plug I (AVP1) has not been licensed for use for closure of patent ductus arteriose. We report our initial experience to occluding special type patent ductus arterioses with the AVP1-a single lobe device of single layer Nitinol mesh for short vessel landing zones.

METHODSPatients referred with small and long Krichenko E patent ductus arterioses 1 mm to 3 mm in diameter underwent occlusion using AVP1. All cases underwent pre-, intra- and post-procedural echocardiography and chest X-ray at the completion of the procedure, the next day and at a 30-day, 3-month and 6-month follow-up visits. Device sizing for device waist diameter and length was based on aortography.

RESULTSFrom April 2008 to June 2012, 26 patients with a mean age of (7.6 ± 8.0) years (range 6 months-32 years) and a mean weight of (23.8 ± 14.8) kg (range 7-67 kg) underwent successful patent ductus arteriose closure. The mean ductus diameter was (2.1 ± 0.7) mm (range 1-3 mm). Transpulmonary (22/26) and transaortic approaches (4/26) were used. No persistent patency was observed after 24 hours and after one month. No device displacement, residual flow and iatrogenic coarctation of the aorta were observed after three months and six months.

CONCLUSIONSThe AVP1 makes it easy to close some Krichenko E patent ductus arterioses. Smaller delivery catheter profile and symmetric cylindrical device shape allow for use for small and long Krichenko E patent ductus arterioses 1 mm to 3 mm in diameter and small patients through transaortic approaches. Broader experience is required to further delineate device and patient selection as well as to document its long-term efficacy and safety.

Adolescent ; Adult ; Child ; Child, Preschool ; Ductus Arteriosus, Patent ; surgery ; Female ; Humans ; Infant ; Male ; Septal Occluder Device ; Young Adult

OBJECTIVETo investigate the efficacy and safety of percutaneous radiofrequency perforation and valvuloplasty in infants with pulmonary atresia with intact ventricular septum (PA/IVS).

METHODSFour infants (body weight 4 - 10 kg) aged 11 months, 9 months, 12 days and 9 months old, respectively, were hospitalized for dyspnea and cyanosis. All patients had a continuous murmur in the left second intercostal space. Doppler echocardiogram showed membranous pulmonary atresia with intact ventricular septum. Right ventriculogram showed a tripartite right ventricle, vasiform infundibulum, and membranous pulmonary valve atresia without ventriculocoronary connections. Descending thoracic aortogram showed good-sized confluent pulmonary arteries being filled from a ductus arteriosus. All the patients were taken up for radiofrequency perforation followed by a balloon dilatation. A 6F Judkins right coronary guiding catheter was positioned in the right ventricular outflow tract and under the atretic pulmonary valve membrane. The radiofrequency perforation catheter along with coaxial injectable catheter was then passed through the right coronary guiding catheter, using it as the guide to the imperforate membrane. The proximal end of the radiofrequency perforation catheter was then connected to radiofrequency generator. After the cusps of pulmonary valve were perforated, the coaxial injectable catheter was moved into the main pulmonary artery. A tiny floppy-tipped coronary guidewire was then passed through the coaxial injectable catheter into the main pulmonary artery and directed through the patent ductus arteriosus into the descending thoracic aorta or directed into pulmonary arteriola. Thereafter, serial balloon dilation catheters were introduced across the pulmonary valve, and dilations were sequentially performed with increasing balloon diameters. The balloon was dilated until the concave of the balloons disappeared. The radiofrequency energy (5 to 8 W) was delivered for 2 to 5 seconds once, but commonly twice, to perforate the valves. After a predilation with a 3 mm x 20 mm to 5 mm x 20 mm balloon at 6 - 14 atm pressure, the valve was subsequently dilated with 10 mm x 30 mm to 14 mm x 30 mm balloon once or twice. The duration of procedures was 120 to 150 min and exposure time was 25.4 to 43.9 min.

RESULTSThe primary procedure was successful in all the infants except one who died early of cardiac perforation with tamponade. After a follow-up period ranging from 2 to 8 months (mean 4.3 m), the remaining 3 survivors achieved complete biventricular circulation. Two of them were awaiting occlusion of the patent ductus arteriosus and 1 needed right ventricular outflow tract reconstruction because of infundibular obstruction.

CONCLUSIONPA/IVS consists of 0.7% to 3.1% of congenital heart defects. 85% of the untreated patients die within half a year. Surgical repair for the infants with PA/IVS is associated with a high mortality. In carefully selected patients with PA/IVS, radiofrequency perforation and balloon dilatation of the pulmonary valve is feasible and may represent a new alternative to surgery due to its low mortality and avoidance of cardiopulmonary bypass.

Balloon Occlusion ; Catheter Ablation ; methods ; Catheterization ; methods ; Female ; Humans ; Infant ; Infant, Newborn ; Male ; Pulmonary Atresia ; physiopathology ; therapy ; Pulmonary Valve ; surgery ; Ventricular Septum

To study the genetic polymorphism of HPA 1-16 platelet antigen alleles among unrelated volunteer donors and establish a typed platelet donor panel in Handan, typing was perfomed by polymerase chain reaction using sequence-specific primers (SSP-PCR); 148 random unrelated blood donors in Handan were genotyped for each of the HPA 1-16 antigen. The gene frequencies were analyzed and the genetype frequencies were determined by direct counting, and these data were compared with HPA distribution among various population by the chi-square test. The results indicated that HPA-1a, 2a, 4a-14a, 16a genes were found among the 16 HPAs in every sample tested. Monomorphic HPA-4a, 7a-14a, 16a were found in the samples. For HPA-1, 2, 5 and 6, a/a homozygosity was predominant with frequencies of 0.9595, 0.8108, 0.9865, 0.9797, respectively, and none of HPA b/b was found in the samples. HPA-1b, 2b, 5b, 6b were rarely found among subjects. HPA-15 had the greatest heterozygosity with a gene frequency of 0.2230, 0.5270, 0.2500 for HPA15a/15a, HPA15a/15b, HPA15b/15b, respectively. HPA-3 showed the second greatest heterozygosity with a gene frequency of 0.3851, 0.5135, 0.1014 for HPA3a/3a, HPA3a/3b, HPA3b/3b, respectively. HPA genotype frequencies showed a good fit to Hardy-Weinberg equilibrium. HPA1-5 gene frequencies for Chinese people in Handan were consistent with those of Chinese people in Shijiazhuang (P > 0.05). Among the HPA1-13, -15, the frequencies of HPA-1, -2, -6 for Chinese people in Handan differed appreciably from those for Chinese people in Taiwan (P < 0.05), others were similar to those of Chinese people in Taiwan. Among the HPA 1 - 8, a similarity was noted between Chinese people in Handan and Koreans (P > 0.05), except for HPA-3. Frequencies of HPA-1, -2, -5 significantly were differed from those in African Americans, as compared with HPA 1-5 (P < 0.05). Comparison of gene frequencies from HPA-1 and -5 showed significant differences between Chinese people in Handan and people in UK (P < 0.05). It is concluded that HPA-2, -3, -5, -15 of people in Western region of China have polymorphism, incompatible frequency of HPA antigen distribution is higher, which inevitably results in the increase of immunologic exposure, therefore attention must be paid to the importance of HPA-2, -3, -5, -15 in clinical disorders. This study for the first time completely analyses HPA1-16 gene frequencies in China, and provides data for establishing a typed platelet donor panel in Handan, China.
Antigens, Human Platelet ; classification ; genetics ; Blood Donors ; statistics & numerical data ; China ; Gene Frequency ; Genotype ; Humans ; Platelet Transfusion ; Polymerase Chain Reaction ; methods ; Polymorphism, Genetic

BACKGROUNDCardiac resynchronization therapy (CRT) device and coronary sinus (CS) lead extraction are required due to the occurrence of systemic infection, malfunction, or upgrade. Relevant research of CS lead extraction is rare, especially in developing countries because of the high cost and lack of specialized tools. We aimed to evaluate percutaneous extraction of CS leads by modified conventional techniques.

METHODSOf 200 patients referred for lead extraction from January 2007 to June 2011, 24 (12.0%) involved CS leads (24 CS leads). We prospectively analyzed clinical characteristics, optimized extraction techniques and feasibility of extraction.

RESULTSComplete procedural success was achieved in 23 patients (95.8%), and the clinical success in 24 patients (100.0%). The leading indication for CS lead extraction was infection (66.7%). Mean implant duration was (29.5 ± 20.2) months (range, 3 - 78 months). Sixteen CS leads (66.6%) were removed with locking stylets plus manual traction by superior transvenous approach. Mechanical dilatation and counter-traction was required to free fibrotic adhesions and extract 4 CS leads (16.7%), which had longer implant duration than other leads ((62.5 ± 12.3) vs. (22.9 ± 14.1) months, P < 0.05). Another 4 CS (16.7%) leads were removed by modified and innovative snare techniques from femoral vein approach. Median extraction time was 11 minutes (range, 3 - 61 minutes) per CS lead, which had significant correlation with implant duration (r = 0.8, P < 0.001). Sixteen patients (66.6%) were reimplanted with new devices at a median of 7.5 days after extraction. Median followed-up was 23.5 months (range, 8 - 61 months), three patients died due to sudden cardiac death (26 months), heart failure (45 and 57 months, respectively).

CONCLUSIONThe modified procedure was proved to be practical for percutaneous extraction of CS leads, especially in developing countries lacking expensive powered sheaths.

Aged ; Cardiac Resynchronization Therapy Devices ; adverse effects ; Coronary Sinus ; surgery ; Device Removal ; methods ; Electrodes, Implanted ; adverse effects ; Female ; Humans ; Male ; Middle Aged ; Prospective Studies

This study was aimed to investigate the molecular genetic basis and serological phenotype of Rh weak D type 15 individuals. Samples were identified by serological tests and genotyped by sequence specific primer-PCR (SSP-PCR), and were sequenced to detect the changes of all ten RHD exons. The number of gene RHD was detected through SSP-PCR. The results showed that in tested individuals of weak D type confirmed by the IAT, 18 cases (56% in weak D) were weak D type 15. Rh factors found in 2 weak D type 15 individuals (11%) were C+c+E+e; Rh factors found in 2 weak D type 15 individuals (11%) were C+c+E-e+; others (78%) were c-c+E+e+. The results by serological tests were consistent with the results genotyped by PCR-SSP method. In all 18 samples, the sequencing result revealed a gene mutation 845G > A at the exon 6 of the RHD and the point mutation changed amino acid G282D of the RhD polypeptide. The zygosity test demonstrated that 2 out of 18 weak D type 15 individuals were RHD(+)/RHD(+) homozygous (two DCe/DcE), 16 cases were RHD(+)/RHD(-) heterozygous (two DCe/dce and fourteen DcE/dce). It is concluded that Weak D type 15 is predominant in weak D individuals of Chinese Han Nationality, and most of them are heterozygous with various RH haplotypes.
Asian Continental Ancestry Group ; genetics ; Base Sequence ; Blood Donors ; China ; ethnology ; Erythrocytes ; immunology ; Exons ; genetics ; Genotype ; Haplotypes ; Humans ; Molecular Sequence Data ; Phenotype ; Point Mutation ; Polymerase Chain Reaction ; methods ; Polymorphism, Genetic ; Rh-Hr Blood-Group System ; genetics ; immunology ; Sequence Analysis, DNA

OBJECTIVETo develop a novel culture system to investigate the effects of ethanol on the function of cultured hepatocytes.

METHODSSandwich configuration was used to culture hepatocytes and the effects of ethanol on functions of bile excretion and protein synthesis as well as the morphology of cultured hepatocytes were observed.

RESULTSBile canaliculi-like structures decreased and anastomatic networks disappeared in ethanol treated hepatocytes. The ability for hepatocytes to internalize, metabolize and excrete compounds into bile was indicated by FDA metabolizing in the hepatocytes. In hepatocytes without ethanol, the bile excretion was showed clearly, but in ethanol-interfered hepatocytes, no bile excretion was observed. After ethanol was given, the level of protein secretion decreased and with the time going, it became lower and lower.

CONCLUSIONHepatocytes can be seriously damaged by ethanol. The study provides a new model to investigate the mechanism of some liver diseases caused by ethanol.

Animals ; Cells, Cultured ; Ethanol ; pharmacology ; Female ; Hepatocytes ; drug effects ; physiology ; Rats ; Rats, Sprague-Dawley

Objective To evaluate the clinical feature of patients with atrial septal defects (ASD)and the safety and efficacy of transcatheter closure of ASD in elderly patients.Methods Between May 2000and June 2010,82 patients aged (64.5 + 3.8)years underwent attempted transcatheter ASD closure.Right heart catheterization was performed before intervention.Echocardiography was made at 1 day,1,3,6 months after the procedure.The pre- and post-closure clinical feature,pulmonary artery pressure (PAP) and cardiac function were evaluated.Results In 82 patients,37 (45.1％) patients were associated with pulmonary arterial hypertension ( PAH ).The systolic PAP and mean PAP [ (44.1 ± 12.4) mm Hg ( 1mm Hg = 0.133 kPa) and (25.2 + 6.8) mm Hg,respectively ] were measured by right heart catheterization before the procedure.One patient was unsuitable for closure because of severe PAH.The remaining 81patients underwent successful ASD closure without major complications.After closuring,systolic PAP decreased from (52.7 ± 10.3 )mm Hg to (31.8 ± 6.3) mm Hg ( P ＜ 0.05 ),and mean PAP descended from (30.9 ± 4.7 ) mm Hg to (21.8 ± 3.4) mm Hg( P ＜ 0.05 ) in the 36 patients with PAH.The cardiac function improved post procedure.There were 6 new-onset atrial fibrillations during follow up.Conclusions ASD in elderly patients are commonly associated with PAH.Transcatheter ASD closure is safe and effective in the majority of elderly patients.

Objective To investigate the clinical significance of gene methylation of p53-bax mitochondrial apoptosis pathway in the carcinogenesis of cholangiocarcinoma. Methods Promoter hypermethylation of DAPK, P14ARF and ASC genes were detected by methylation-specific PCR. Exon 5-8 of p53 gene were examined by automatic sequencing. Results It was found that 66.7% of 36 cholangiocarcinoma patients had methylation of at least one tumor suppressor gene. The rate of tumor suppressor gene methylation in these cholangiocarcinomas was 25.0% in P14FRF, 30.6% in DAPK and 36.1% in TMS1/ASC. The methylation rate of tumor suppressor gene in tissues adjacent to the cancer tissue was 13.9% including 5.6% in DAPK and 8.3% in TMS1/ASC. p53 gene mutation was detected in 22 of 36 patients(61.1%). Fourteen patients (38.9%)was found to have p53 gene mutation associated with the methylation of tumor suppressor gene. The rate of p53 gene mutation and methylation of tumor suppressor gene were statistically and significantly correlated with the features of tumor biology including differentiation and invasion (P< 0.05). Conclusion DNA methylation of p53-bax mitchondrial apoptosis pathway may be a frequent molecular event in the carcinogenesis of cholangiocarcinoma. Although the methylation rate of ASC, DAPK genes is relatively low, it may be helpful for early diagnosis, p53 gene mutation associated with the methylation of tumor suppressor genes may be correlated with tumor malignant biologic features.

Objective To investigate the clinical significance of gene methylation of p53-bax mitochondrial apoptosis pathway in the carcinogenesis of cholangiocarcinoma. Methods Promoter hypermethylation of DAPK, P14ARF and ASC genes were detected by methylation-specific PCR. Exon 5-8 of p53 gene were examined by automatic sequencing. Results It was found that 66.7% of 36 cholangiocarcinoma patients had methylation of at least one tumor suppressor gene. The rate of tumor suppressor gene methylation in these cholangiocarcinomas was 25.0% in P14FRF, 30.6% in DAPK and 36.1% in TMS1/ASC. The methylation rate of tumor suppressor gene in tissues adjacent to the cancer tissue was 13.9% including 5.6% in DAPK and 8.3% in TMS1/ASC. p53 gene mutation was detected in 22 of 36 patients(61.1%). Fourteen patients (38.9%)was found to have p53 gene mutation associated with the methylation of tumor suppressor gene. The rate of p53 gene mutation and methylation of tumor suppressor gene were statistically and significantly correlated with the features of tumor biology including differentiation and invasion (P< 0.05). Conclusion DNA methylation of p53-bax mitchondrial apoptosis pathway may be a frequent molecular event in the carcinogenesis of cholangiocarcinoma. Although the methylation rate of ASC, DAPK genes is relatively low, it may be helpful for early diagnosis, p53 gene mutation associated with the methylation of tumor suppressor genes may be correlated with tumor malignant biologic features.