Hepatitis C virus ( HCV) infection is one of the main reasons causing liver cirrhosis and hepatocellular carcinoma. The quasispecies composition and the evolution of HCV are very complicated. An in-depth analysis of the quasispecies composition of HCV is critical for elucidating the mechanisms of HCV transmission. The regions encoding the envelope glycoproteins (E1 and E2) and the nonstructural protein 2 (NS2) are hyper variable regions (HVR). Analyzing the quasispecies of HCV HVR with advanced sequen-cing and bio-information technologies would be beneficial for understanding the sources of HCV infection, the routes of transmission and the evolution of HCV. Currently, three generations of sequencing methods and some bio-information soft-wares are available for analyzing HCV HVR quasispecies. When an outbreak of HCV infection happens, using the new generation of sequencing and bio-information methods to seek the first case and the routes of transmission would provide scientific basis for the prevention and treatment of HCV in-fection.

Objective:To characterize the CD4+CD45RA+,CD4+CD45RO+,CD8+CD45RA+ and CD8+CD45RO+T lymphocyte subsets in the peripheral blood of the patients with chronic hepatitis B and to explore their relations with the disease state.Methods:The peripheral blood was collected from 104 patients with chronic hepatitis B and 30 healthy individuals,then CD4+CD45RA+,CD4+CD45RO+,CD8+CD45RA+ and CD8+CD45RO+T lymphocyte subsets were detected by flow cytometry with three-color fluorescence technology.Results:To compare with the control,the percent of CD8+CD45RA+T lymphocyte in patients with mild and severe grad CHB decreased markedly,the percent of CD8+CD45RO+T lymphocyte increased markedly,CD4+,CD8+,CD4+CD45RA+ and CD4+CD45RO+T lymphocyte did not change obviously. Compared with the mild grade CHB,the percent of CD8+CD45RA+T lymphocyte in patients with severe grade CHB decreased obviously(P

Objective To determine sonographic features and outcome of fetal lung abnormal lesions. Methods Blood supply of the lesion, fetus hydrops and other extra-lung anomalies should be evaluated by color Doppler flow image(CDFI) when abnormal lesion was detected in fetus lung during routine scanning. The fetus with lung abnormal lesion without hydrops at the first time scanning should be monitored by ultrasound in every four to six weeks. Results Forty cases fetus presented lung abnormal lesions, which included 21 cases with hyperechogenic solid masses,15 cases with cystic-solid mixture masses,4 cases with cystic masses. Lesions of 8 cases were demonstrated systemic arterial blood supply arising from the aorta on CDFI and the diagnosis of pulmonary sequestration (PS) were suggested. Seventeen cases lesion that had small size or decreased or disappeared or remained stable in size with gestational age developing had normal neonate. Conclusions CDFI is very useful in detecting abnormal lesion of the fetal lung and differentiating pathology and evaluating the prognosis. The outcome of isolated lung lesion without hydrops and mediastinal shift that decreased or disappeared or remained stable in size had a good prognosis.

BACKGROUNDTo study to immunogenicity of recombinant HEV ORF2 protein expressed in pichia pastoris.

METHODSBALB/c mice were immunized with the recombinant HEV ORF2 protein. The ability of antiserum to bind HEV was tested using affinity-capture reverse transcription polymerase chain reaction (RT-PCR). Moreover, the recombinant protein was used to immunize BALB/c mice by different routes with different adjuvants. Serum conversion rate of anti-HEV antibody and the ELISA titer were detected.

RESULTSThe antiserum could capture native HEV for RT-PCR. As to the immunization effect, the immune response by intramuscular route was better than that of the intraperitoneal route. The protein with alum and CpG adjuvant could elicits more significant immune responses than using the alum adjuvant alone. The best way was to immunize with the protein with alum and CpG adjuvant by intramuscular route with a boosted injection on the 4th week after the first immunization. The ED50 was 0.023 microgram. This is the first report that the antibody elicited by recombinant HEV ORF2 protein expressed in pichia pastoris recognizes native HEV. High immunogenicity of this kind of ORF2 was also demonstrated by inducing strong immune response in mice with good ED50 result.

CONCLUSIONSThe high immunogenicity of this kind of HEV ORF2 may make a foundation for the development of new type of hepatitis E vaccine.

Animals ; Antibody Formation ; Female ; Hepatitis E virus ; immunology ; Mice ; Mice, Inbred BALB C ; Pichia ; genetics ; Recombinant Proteins ; genetics ; immunology ; Reverse Transcriptase Polymerase Chain Reaction ; Viral Proteins ; immunology

Objective To study the ultrasonographic clues and methods for fetal anomalies of the aorta arch and improve prenatal detection of anomalies of the aorta arch.Methods One thousand four hundred and seventy-two cases fetus who were carried out detailed scan and whose results were confirmed were chose as study objects.Every routine fetal echocardiography included four chamber and left and right outflow tract and three-vessel trachea view(3VT).The more views which included longitudinal and coronary view of the aorta arch and coronary view of the trachea and main bronchus were obtained when the abnormality of aorta arch was suspected.Results One hundred and forty-eight cases with anomalies of aorta arch were diagnosed by ultrasonography.One case was misdiagnosed.Ninety-two fetus with anomalies of aorta arch which included 28 aortic coarctation(CoA) and 10 interrupted aortic arch (IAA) and 52 right-side aortic arch and abnormal aortic branch and 2 double aortic arch were confirmed by postmortem or postnatal echocardiography and surgery.Of the 92 confirmed cases,24 had prenatally diagnosed additional complex intracardiac anomalies.All cases with CoA and IAA presented ventricular and/or great arterial disproportion with smaller left ventricle and aorta diameter on four chamber view and 3VT.Right aortic arch (RAA) and abnormal aortic branch(AAB) displayed aortic arch located on the right side of the trachea and increased distance between the aortic arch and arterial duct and abnormal aortic arch branch-subclavian artery originating from the beginning section of the descend aorta which coursed behind the trachea with U-shaped appearance on the 3VT plane.The display rate of the transverse and longitudinal and coronary view of the aorta arch was 98.4%,90.0%,81.9%,respectively.Conclusions Disproportional ventricular and /or great arterial with smaller left ventricle and aorta diameter are the clues for CoA and IAA.Increased distance between the aortic arch and arterial duct is the clue for RSA.The transverse view of the aortic arch 3VT is the most sensitive for detecting the anomalies of the aortic arch and the most easily be obtained.The longitudinal and coronary view of the aorta arch and coronary view of the trachea and main bronchus are helpful in differentiating the anomalies of the aortic arch.

Objective To study the ultrasonic features and outcomes of VACTERL association fetuses. Methods From Jan 2003 to Sep 2013 in Shenzhen Maternity&Child Healthcare Hospital there were 55 fetuses diagnosed as VACTERL association, the prenatal ultrasonographic characteristics and outcomes were summarized. Results Ultrasonography showed that the incidence of 6 VACTERL association anomalies were:vertebral defects were 45.5%(25/55), anal atresia were 30.9%(46/55), cardiac malformations were 81.8%(45/55), tracheoesophageal ifstula/esophageal atresia were 32.7%(18/55), renal anomalies were 60.0%(33/55) and limb anomalies were 83.6%(17/55). All the 55 fetuses had 3 or more VACTERL association malformations and the characteristic ultrasonic features were as follows:(1) There were 39 cases (70.9%, 39/55) co-occur with three VACTERL malformations, the more common malformations were limb anomalies (33/39), cardiac malformations (31/39) and renal anomalies (21/39). (2) There were 13 cases (23.6%, 13/55) co-occur with four VACTERL malformations, the more common malformations were cardiac malformations (11/13), limb anomalies(10/13), renal anomalies (9/13) and tracheoesophageal ifstula/esophageal (8/13). (3) There were 3 cases (5.5%, 3/55) co-occur with ifve VACTERL malformations, they were all with anal atresia, cardiac malformations, renal anomalies and limb anomalies (3/3), and two were co-occur with vertebral defects (2/3). (4) No fetus co-occur with six VACTERL malformations. There were 29 cases (52.7%, 29/55) co-occur with other malformations, of which 21 cases (38.2%, 21/55) with single umbilical artery. All the 55 cases underwent labor induction. The results of postmortem appearance of 55 cases and autopsies of 9 cases were all consistent with prenatal ultrasound scan. Conclusions Fetuses with VACTERL association had characteristic prenatal ultrasound imaging, multiple malformations can be found and limb anomalies, cardiac malformations and renal anomalies are more common. Scanning the fetal structures from higher incidence to lower incidence of VACTERL association can be helpful to improve the diagnostic coincidence rate of VACTERL association. Prenatal ultrasound diagnosis of VACTERL association can provide guidance for clinical obstetrical management.

Objective To summarize and analyze prenatal ultrasound and postnatal autopsy ifndings in fetuses with urorectal septum malformation sequence (URSMS). Methods An analysis of prenatal ultrsound ifndings and postnatal autopsy features was performed on eleven cases of fetuses with URSMS that were identiifed by ultrasonography at Shenzhen Maternity&Child Healthcare Hospital in the period of January 2003 to December 2012. Results Prenatal ultrasonography showed a large abdominal cystic mass concomitant with imperforate anus in eleven fetuses with URSMS. The cyst contained unilocular or bilocular cystic structures in two fetuses, and trilocular cystic structures in nine fetuses. The cyst was demonstrated as clear acoustic transmission in three fetuses and unclear in eight fetuses. Out of them, seven fetuses had kidney abnormalities, six had ascites, and three had enterolithiasis. The associated systemic abnormalities included tethered cord in two fetuses, single umbilical artery in two fetuses, sacrococcygeal dysplasia in one fetus, and myocardial noncompaction in one fetus. 21-trisomy was found in one fetus by chromosome examination. Eleven cases were all identiifed as female fetuses by autopsy ifndings, including a single perineal opening and ambiguous genitalia with clitoral hypertrophy and labial fusion. The internal genital abnormalities included double vagina or longitudinal vaginal septum in nine fetuses, double uterus or uterus bicornis in ten fetuses and vaginal dysplasia in one fetus. Conclusions URSMS is a complex congenital malformation, which includes abnormalities of the urinary system, reproductive system and gastrointestinal track. An abdominal cystic mass visualized by prenatal ultrasonography might be the distinctive lesion in female with URSMS, and have an important diagnostic value. The kidney abnormalities and ambiguous genitalia can contribute to the diagnosis of URSMS.

ObjectiveTo summarize prenatal ultrasound image features of Cantrell′s syndrome (Cantrell′s pentalogy).MethodsFrom January 2007 to December 2013 in Shenzhen Maternity & Child Healthcare Hospital, there were twenty cases diagnosed and conifrmed by postmortem as Cantrell′s syndrome. The prenatal ultrasound image features and outcomes were analyzed.ResultsAmong the twenty cases, eighteen were singleton and two were twins with the other normal fetus. Prenatal ultrasound image: two obvious ultrasound image features of ectopic heart and hight omphalocele (seventeen cases) or gastroschisis (three cases) were appeared in prenatal ultrasound of twenty cases. Fourteen cases were complete ectopic heart (Ectopic heart was located within the omphalocele in one case, ectopic heart was completely exposed in amniotic lfuid in thirteen cases), six cases were incomplete ectopic heart, three cases were ventricular septal defect in which one was associated with pulmonary stenosis, two cases were single atrium and single ventricle in which one was associated with a single arterial trunk, one case was tetralogy of Fallot, and six cases were without signiifcant heart abnormalities. Associated with other abnormalities: seventeen cases were associated with other abnormalities in twenty cases, in which eleven cases were spinal abnormalities, nine cases were umbilical cord abnormalities (short umbilical cord in six cases, single umbilical artery in two cases, short umbilical cord associated with single umbilical artery in one case), ifve cases were neurological abnormalities (exencephalia in four cases, encephalocele in one case), four cases were limb abnormalities, three cases were cheilopalatognathus, one case was giant bladder. Three cases weren′t associated with other abnormalities. Nuchal Translucency: in eight cases, nuchal translucency were measured in early pregnancy. Nuchal translucency of ifve cases were thick, one case was cervical hygroma in one of the two twins. Chromosome examination results: only three cases had chromosome examination in twenty cases, in which one case was trisomy 21, another two were chromosome normal. Fetal outcome and examination results: in twenty cases, one of the two twins can′t survive after birth, and the other one of two twins was reduced in intrauterine, so the specimen could not be obtained. Eighteen cases of singleton underwent labor induction. The results of postmortem appearance (nineteen cases) and autopsies (ifve cases) were all consistent with prenatal ultrasound scan.ConclusionsEctopic heart and hight omphalocele or gastroschisis appearing at the same time was the characteristic of Cantrell′s syndrome in prenatal ultrasound and other abnormalities may be part of the Cantrell′s syndrome spectrum. It is beneift to detect and diagnose Cantrell′s syndrome by prenatal ultrasound in the ifrst trimester.

OBJECTIVETo obtain recombinant baculovirus containing full-length structural gene of HEV and to express HEV structural protein.

METHODSFull-length structural gene of HEV (5147-7126 nt) was inserted into baculovirus expression vector pAcUW51. The recombinant plasmid and Baculo Gold DNA were co-transfected into insect cell line Sf9. Single plaque was picked and amplified and expression of HEV structural protein was tested by SDS-PAGE, Western blot and immunofluorescence methods.

RESULTSSDS-PAGE analysis showed HEV structural protein was highly expressed; Western blot and immunofluorescence assay showed that the expression product could specifically react with HEV positive sera, confirming the protein possessing HEV specific antigenicity.

CONCLUSIONSHEV structural protein was successfully expressed using baculovirus expression system.

Animals ; Baculoviridae ; genetics ; Cells, Cultured ; Gene Expression ; Genes ; genetics ; Genetic Vectors ; Hepatitis E virus ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; Viral Structural Proteins ; biosynthesis ; genetics

Objective To investigate the clinical value of acoustic radiation forcs impulse (ARFI)in quantitative evaluating placental elasticity.Methods The study population included 487 normal pregnant women,and ARFI generated shear wave velocity (SWV)was measured.On the basis of gestational ages, placental sites and sampling depths in region of interest (ROI),the normal population was divided into different groups.One-way analysis of variance was used to compare the discrepancy on the SWV values amomg the nomal placental sites or the sampling depths in ROI.Pearson correlation coefficient were used to assess the possible relationships between the normal placental SWV values and the different gestational ages or the placental grades.A total of 5 1 cases were diagnosed with severe preeclampsia,among them 25 cases were categorized as severe preeclampsia with fetal growth restriction (FGR).The placental SWV values were measured and compared with those of the normal population.The pathological examinations were performed on 50 normal and 5 1 abnormal placentas.Results Basing on the placental sites,the normal population were divided into three groups:anterior wall,lateral wall and posterior wall groups.Compared with the posterior wall group,the placental SWV values in the anterior wall or lateral wall group significantly decreased (P <0.05).No statistical significant difference was found between the anterior wall group and the lateral wall group.Basing on the distance from the sampling depths to the probe (range from 2 to 7.99 cm,and each additional 1 cm corresponding a group),the normal population were divided into six groups.There was statistical significant difference for the placental SWV values between the two groups of distance in the range of 2.0 to 5.99 cm and 6.0 to 7.99 cm (P <0.05).There was no statistical significant difference between the other two groups.The mean SWV value was (0.78±0.08 m/s)in the normal group. No significant relationships were found between the placental SWV values and the gestational ages of the normal population.However the placental SWV values were significantly related to the placental grades.For the placental SWV values,there was statistical significant difference between the normal group and the abnormal group (P <0.05).No statistical significant difference was found between the severe preeclampsia group and the severe preeclampsia with FGR group.The pathological examinations also showed significant changes in the abnormal group.Conclusions ARFI may quantitatively analyze the placental elasticity and make a difference between nomal and abnormal placenta.