Autoantibodies ; Child ; Complement Factor H ; immunology ; Female ; Hemolytic-Uremic Syndrome ; immunology ; Humans

Objective To investigate the immunogenicity and cross protective effects of two novel HCV DNA vaccines in a mice model.Methods Two self-replicating alphavirus vector-based HCV DNA vaccines, pSCK CE1E2Y and pSCK H155, were constructed based on the genes encoding the structural pro-teins (Core, E1 and E2) and structural and NS3 fusion proteins (Core, E1 , E2 and NS3) of a HCV strain isolated from a Chinese patient (genotype 1b, Hebei strain), respectively.Western blot analysis was per-formed to detect the expression of fusion antigens.The BALB/c mice were intradermally immunized with the recombinant DNA vaccines by using electroporation.The immune responses induced in mice and the cross protective effects of the recombinant DNA vaccines were evaluated.Results The DNA vaccines effectively expressed the target antigens in vitro.The antigen-specific antibody responses and specific T cell immune re-sponses were induced in mice by the immunization of replicative DNA vaccines.However, no effective cross protection was provided by either of the DNA vaccines in the surrogate challenge model based on a recombi-nant heterologous HCV (JFH1, 2a) vaccinia virus strain.Conclusion Although no effective cross protec-tion was observed, both of the two replicative DNA vaccines could induce strong humoral and cellular im-mune responses against multi-target antigens of HCV strains.This study has paved the way for further inves-tigation on the development of novel HCV vaccines.

Objective To investigate the development strategy of novel T cell based vaccine against HCV infection.Methods BALB/c mice were primed with pSCK-based DNA vaccine and boosted with type 5 adenoviral vector-based vaccine, which expressed the structural proteins ( Core, E1 and E2) de-rived from a Chinese HCV patient (genotype 1b, Hebei strain).Enzyme linked immunospot assay (ELIS-POT) and intracellular cytokine staining ( ICS) were used to analyze the elicited antigen-specific immune re-sponses and the efficacy of cross-protection.Results Immunization of mice with the prime-boost vaccination strategy elicited stronger T cell immune responses against multiple HCV antigens than using the DNA vac-cines alone, especially the IFN-γ-secreting T cell responses against E1 protein as indicated by ELISPOT as-say.ICS data indicated that the prime-boost regimen elicited more TNF-α-producing CD4+and IFN-γ-produ-cing CD8+T cells against E1 protein and high levels of IFN-γ-producing CD4+and CD8+T cells against E2 protein in comparison with immunization with DNA vaccines.Moreover, the prime-boost vaccination was ca-pable of eliciting effective cross-protection in a surrogate challenge model based on a recombinant heterolo-gous HCV (JFH1, 2a) vaccinia virus.Conclusion The prime-boost vaccination using DNA and rAd5-based vaccine expressing HCV structural antigens induced significant cellular immune response and cross-protection in mice, suggesting the possibility of using it as a promising T cell based vaccine against HCV in-fection.

OBJECTIVETo compare the short-term clinical outcome between unilateral fixation fusion (ULF) and minimally invasive spine transforaminal lumbar interbody fusion (MIS-TLIF) in treating lumbar disc herniation (LDH).

METHODSThe clinical data of 39 patients with LDH were retrospectively analyzed from June 2008 to March 2013. There was 22 males and 17 females, aged from 45 to 75 years old with an average of 56.9 years. Therer were 3 cases in L3,4, 15 cases in L4,5, 21 cases in L5S1. Among them, 21 patients underwent unilateral fixation fusion (ULF group) and 18 underwent minimally invasive spine transforaminal lumbar interbody fusion (MIS-TLIF group). Operation time, blood loss, the times of radiographic exposure and hospital stay were noted and compared between two groups. Radiograph informations were regularily accessed and VAS, ODI scores were recorded at 3 days and 3, 6, 12 months after operation, respectively. According to modified Macnab criteria, the clinical effects were evaluated at final follow-up.

RESULTSAll operations were successful without severe complications. The averaged operative time and the times of radiographic exposure in ULF group [(95 ± 25) min and (4.2 ± 0.4) times] were less than that of MIS-TLIF group [(120 ± 35) min and (10.1 ± 3.9) times] (P < 0.05). But, the mean blood loss and hospital stay in MIS-TLIF group [(75 ± 45) ml and (7.2 ± 2.2)d ]were less than that of ULF group [(165 ± 60) ml and (11.0 ± 3.7) d] (P < 0.01). All patients were followed up from 12 to 45 months with an average of 29.5 months. The VAS and ODI score had significantly improved during the follow-up and no significant differences were found between two groups at the same time point (P > 0.05). The postoperative radiographs showed internal fixation position was good. And all patients obtained bone fusion by CT scan at 1 year after operation. There was no significant differences in modified Macnab criteria between two groups at the latest follow-up (P > 0.05).

CONCLUSIONFavorable short-term clinical effects can be achieved in suitable LDH patients with ULF or MIS-TLIF surgical procedures.

Aged ; Humans ; Intervertebral Disc Displacement ; surgery ; Lumbar Vertebrae ; surgery ; Middle Aged ; Minimally Invasive Surgical Procedures ; methods ; Spinal Fusion ; methods

Objective To characterize the immunogenicity in gene immunization of the conserved regions of hepatitis C virus(HCV) based on different delivery strategies. Methods We first constructed a novel DNA vaccine encoding a fusion gene(from partial NS3 and Core) of HCV. Then we compared different protocols based on naked DNA injection twice or DNA injection with gene electrotransfer(GET) in BALB/c mice. The immune response was measured by antibody ELISA and by IFN-gamma ELISPOT. Results Our data showed that a protocol based on intradermally injection of DNA with optimal GET induced the strongest humoral and cellular immunity, and DNA with GET induced a substantially higher anti-NS3/Core T cell re-spoase than naked DNA injection. Conclusion Our data suggest that DNA vaccines encoding NS3/Core fu-sion protein of HCV immunized by the present strategy could merit further study in the context of future prophylactic and therapeutic HCV T cell based vaccines.

Objective To develop an effective and broad immune protective H5N1 vaccine.Methods We first developed two recombinant vaccinia ( Tiantan strain) virus ( rTTV ) based H5N1 vaccines, which consisted of bicistron expressing the hemagglutinin(HA) and matrix protein 2(M2), or bicistron expressing the neuraminidase(NA) and matrix protein 1 (M1). The expression of H5N1 protein in rTTVs was confirmed. We immunized the BALB/c mice twice with two kind of dose ( 104 PFU, 107 PFU)using different combination. Subsequently, we assessed the humoral and cellular immune response in vaccinated mice. Results Our data showed that rTTV-based H5N1 vaccine induced rapidly robust HA- and NAspecific antibody level and IFN-γ secreting form cell(SFC) with either single dose of 107 PFU or twice dose of 104 PFU or 107 PFU. We also detected significant neutralizing antibody and matrix-specific immune response. In addition, we found that immunization with two kind of rTTV-based H5N1 vaccines induced much high level of M2-specific antibody than that with single of rTTV-based H5N1 vaccine. Conclusion rTTVbased H5N1 vaccines in this study elicited board array of immunity and our study offers a promising alternative H5N1 vaccine candidates with favorable potential to prevent various H5N1 pandemic.

OBJECTIVETo study the polysaccharide of Condonopsis pilosula.

METHODThe polysaccharide, CPP-1, was purified by DE-52 cellulose and Sephadex G-200 gel column chromatography. Purity and molecular weight of the polysaccharide were determined by gel permeation chromatography. Methylation analysis, periodate oxidation and degradation, IR, 1H-NMR and 13C-NMR methods were adopted to elucidate the chemical structure.

RESULTThe molecular weight of CPP-1 was estimated to be 7.5 x 10(4), and the structure of CPP-1 was a beta-(2 --> 1) linked beta-D-fructosan.

CONCLUSIONCPP-1 was a neutral homosaccharide.

Codonopsis ; chemistry ; Drugs, Chinese Herbal ; chemistry ; Fructans ; chemistry

The membrane of sodium cellulose sulphate ( NaCS)-poly dimethyldiallylammonium chloride (PDMDAAC) microcapsule is compact and has low molecular weight cut-off, which would delay the mass transfer and affect the cell growth immobilized in the capsule. Macroporous NaCS-PDMDAAC microcapsules were prepared using the degradation of the starch by amylase in the membrane of the capsules. The pore size and the permeability in the membrane were improved obviously. As model cells, the Candida krusei CK1 and E. coli EC1 immobilized in the capsules were cultured in the shake flask and bubble column respectively. It was shown that the cell density immobilized in the microcapsules cultured in the bubble column was higher than that cultured in the shaking flask. It implied that the limiting factor of the cell growth in the capsule lied in the diffusion of the oxygen. Since the rate of the oxygen transporting across the membrane was greatly enhanced due to the enlarged pore size, the maximum cell density in the macroporous capsules was 20%-110% over than that in the standard capsules in the bubble column. However, the extent of E. coli cell density increasing was higher than that of the yeast, which may be due to the difference of the oxygen requirement between the two microbes.
Amylases ; metabolism ; Bioreactors ; Candida ; growth & development ; Capsules ; chemical synthesis ; chemistry ; Cell Culture Techniques ; methods ; Cells, Immobilized ; Cellulose ; analogs & derivatives ; chemical synthesis ; chemistry ; Escherichia coli ; growth & development ; Membranes, Artificial ; Polyethylenes ; chemical synthesis ; chemistry ; Porosity ; Quaternary Ammonium Compounds ; chemical synthesis ; chemistry ; Sodium ; Surface Properties ; Temperature ; Time Factors

OBJECTIVEAnti-neutrophil cytoplasmic antibody (ANCA)-associated vasculitis is a disorder with poor prognosis. This study aimed to improve the diagnosis and treatment of ANCA associated vasculitis of children, to analyze the clinical features, pathological characteristics and the prognosis of children with ANCA-associated vasculitis.

METHODFifteen children with ANCA associated vasculitis who were hospitalized from 2003 to 2012 in our hospital were included. Their data of pre-diagnosis status, clinical manifestations, renal pathology, treatment and prognosis were reviewed retrospectively.

RESULTOf the 15 children, 11 were girls and 4 boys with a mean age of 10.7 years. Fourteen children were categorized as microscopic polyangitis. The time to diagnosis varied from 0.5 month to 40 months. Hematuria and proteinuria were revealed by urine analysis in all of them, only 6 children complained with gross hematuria or edema of oliguria. Decreased glomerular filtration rate was revealed in 13 children, 8 of whom had a creatinine clearance rate of less than 15 ml/(min·1.73 m(2)). Twelve children underwent renal biopsy, crescent formation was found in 11 children. Most of the crescents were cellular fibrous crescents or fibrous crescents. Six children were diagnosed as crescentic nephritis; the process of rapidly progressive nephritis was only observed in 2 children. Segmental glomerulosclerosis or global glomerulosclerosis were found in 10 children, 3 of them were diagnosed as sclerotic glomerulonephritis. Anemia and pulmonary injury were the most common extra renal manifestations. Other extra renal manifestations included rash, pain joint, gastrointestinal symptoms, abnormal findings of cardiac ultrasonography and headache. Eight children were treated with steroid combined with cyclophosphamide, 4 were treated with steroid and mycophenolate mofetil, 2 were treated with steroid, cyclophosphamide and mycophenolate mofetil, 3 children were treated with plasma exchange. Fourteen children were followed up for 0.5 month to 4 years. The renal function did not recover in children with creatinine clearance rate of less than 30 ml/(min·1.73 m(2)), who showed crescentic glomerulonephritis or sclerotic glomerulonephritis. The children who had creatinine clearance rate of more than 30 ml/(min·1.73 m(2))had better prognosis.

CONCLUSIONMore attention should be paid to ANCA-associated vasculitis among school age girls with anemia or pulmonary diseases. The renal damage was serious in children; however, the clinical manifestations were not obvious. Children with a creatinine clearance rate of less than 30 ml/(min·1.73 m(2)) had poor prognosis. Early accurate diagnosis is very important.

Adolescent ; Anemia ; etiology ; pathology ; Anti-Neutrophil Cytoplasmic Antibody-Associated Vasculitis ; complications ; diagnosis ; pathology ; Antibodies, Antineutrophil Cytoplasmic ; blood ; immunology ; Biopsy ; Child ; Child, Preschool ; Creatinine ; blood ; Female ; Glomerulonephritis ; pathology ; Hematuria ; etiology ; pathology ; Humans ; Kidney ; pathology ; physiopathology ; Kidney Function Tests ; Male ; Nephritis ; diagnosis ; etiology ; pathology ; Prognosis ; Proteinuria ; etiology ; pathology ; Retrospective Studies

Objective To investigate the etiology composition of end-stage renal disease (ESRD) in children,in order to provide reference for the prevention and treatment.Methods The children with ESRD who were diagnosed in Peking University First Hospital from January 2005 to October 2013 were selected,and the etiology composition and incidence of the children with ESRD were retrospectively analyzed.Diagnostic criteria for children with ESRD refer to the clinical practice guidelines for chronic renal disease (NKF/KDOQI),developed by the American kidney foundation in 2002.Results Eighty-six children with ERSD were enrolled including 53 cases of males,33 cases of females,with the male to female ratio of 1.61 ∶ 1.00 and the mean onset age was (7.08 ± 4.23) years old,and their average diagnosis age was(9.25 ±4.17) years old.The median duration of ERSD before diagnosis was 0.84(0.01-13.67)years.The main cause of ESRD was acquired renal disease,accounting for 43.02％ (37/86 cases),mainly the chronic glomerulonephritis (18/86 cases,20.93％) and nephrotic syndrome (16/86 cases,18.60％);followed by urinary congenital abnormity,accounting for 40.70％ (35/86 cases),in which the most common were renal dysplasia (18/86 cases,20.93％) and cystic renal disease (11/86 cases,12.79％).Children under 3 years old mainly showed congenital urinary tract abnormalities(6/10 cases,60.00％).But children over 3 years old mainly showed acquired renal diseases (37/76 cases,48.7％),and pathologic classification of glomerular disease were proliferative mesangial glomerulonephritis (6/23 cases,26.09％),focal segmental glomerulosclerosis (5/23 cases,21.74％) and interstitial nephritis(3/23 cases,13.04％).Conclusions The main etiology of ESRD is glomerular disease and congenital abnormal development of urinary system,therefore,more attention should be paid on the ultrasound screening of the urinary tract in the perinatal period and urine screening in children.There are great significances in reducing the incidence of ESRD and intervening actively the progression to chronic kidney disease.