In order to establish the optimum condition for purifica tion of the nucleoprotein(NP) of rabies virus by immunoaffinity chromatography, the efficient and non-denaturative eluents(Mg-el) was obtained by using ELISA elution model; furthermore, it didn't damage the activity of NP. Two kind of NPs , expressed by recombinant vaccinia virus (rVac-N) and recombinant baculovirus (BRN), were purified by a Sepharose CL 4B column and a 2C12- Sepharose 4B colum n. By Western-blot and SDS-PAGE, high purity and good antigenical intact NPs w ere identified. The purified ribonucleoprotein (RNP) of rabies virus 5aG strain was also obtained. After immunized with NP and RNP, mice developed a strong anti -nucleoprotein response and were protected against a lethal challenge of rabies virus CVS strain. There were not difference been observed among the mice immuni zed with different purified protein. These data indicate that the NPs are antige nical and immunogenical comparable to the authentic rabies RNP and therefore pre sent a potential source of an effective ,safe and economical subunit vaccine.

Animals ; Carps ; genetics ; Cloning, Molecular ; Fish Proteins ; genetics ; Hydroxylation ; Hypoxia-Inducible Factor 1, alpha Subunit ; genetics ; Lakes

OBJECTIVETo investigate magnetic resonance imaging(MRI) manifestations of perianal abscess and to evaluate MRI diagnosis for perianal abscess.

METHODSFifty cases of perianal abscess between July 2007 and March 2009 were included in this study. MRI was performed using T1 weighted sequence in axial plane, T2 weighted sequence with fat saturation in axial, coronal and/or sagittal plane, and T1 weighted enhanced sequence with fat saturation in axial, coronal and/or sagittal plane. The location, size, shape, signal characteristics, and enhancement patterns of perianal abscess were analyzed.

RESULTSFifty cases with 51 perianal abscesses were identified. The abscess appeared slight hypo- or isointense signal on T1 weighted imaging, obviously hyperintense signal at cavity and isointense signal at wall on T2 weighted with fat saturation imaging, and marked enhancement at wall. The largest diameters ranged between 0.5 cm and 9.0 cm (mean 3.4±1.7 cm ) in abscess, between 0.2 cm and 8.0 cm(mean 2.7±1.7 cm) in cavity. The abscess was round, oblong and crescent in 23, 18 and 7 cases respectively, and was unilocular in 41 cases and multilocular in 10 cases. The abscess located between levator ani and intersphincteric groove in 23 cases, under intersphincteric groove in 3 cases, across levator ani and intersphincteric groove in one case, on levator ani in 7 cases, on intersphincteric groove in 16 cases and above levator ani in one case.

CONCLUSIONMRI is a noninvasive, convenient and highly accurate procedure for diagnosing perianal abscess, which can demonstrate the anatomical relationship between abscess and anal canal.

Abscess ; diagnosis ; pathology ; Adolescent ; Adult ; Aged ; Anus Diseases ; diagnosis ; pathology ; Child ; Female ; Humans ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Retrospective Studies ; Young Adult

OBJECTIVETo observe the association between angiotensin-converting enzyme (ACE) gene polymorphism and endothelial nitric oxide synthase (eNOS) gene polymorphism and risk of coronary artery disease (CAD) in Han Chinese.

METHODSThe polymorphism in the ACE and eNOS gene were detected by using polymerase chain reaction-restriction fragment length polymorphism analysis, blood pressure (BP), blood lipids, blood glucose (BS), body mass index (BMI) and left ventricle eject fraction (LVEF) were determined 236 patients with CAD and 190 healthy individuals.

RESULTSThe frequencies of DD genotype of ACE were higher and the II genotype were lower in CAD patients than in controls (P < 0.05). CAD patients with DD genotypes were related with higher serum TG, lower HDL-C, higher BS levels, higher BWI and lower LVEF compared to CAD patients with II and ID genotypes of ACE (all P < 0.05), while SBP, DBP, TC and LDL-C levels were similar among CAD patients and controls with different genotypes of ACE (P > 0.05). The genotype distributions of ACE and eNOS were also similar among CAD patients with or without diabetes mellitus/ACS, with single or multiple vessel diseases (P > 0.05). The frequency of GT genotype of eNOS was higher in CAD patients than in controls (P < 0.01) while the frequency of GG genotype in CAD patients and controls was similar (P > 0.05) and eNOS genotypes were not related to TC, TG, HDL-C, LDL-C, BS, BMI, SBP, DBP and LVEF levels among CAD patients and controls (P > 0.05). The risk of suffering from CAD in population with ACE DD genotype is 1.74 times higher than that with II genotype (P < 0.01) and 1.73 times higher in population with eNOS GT genotype than that with GT genotype (P < 0.05). The risk of suffering from CAD is 37.9% with II and GG genotypes and 77.8% with DD and GT genotypes.

CONCLUSIONThe ACE and eNOS genotype polymorphisms were associated with risk of CAD and persons with DD and GT genotypes take higher risk of suffering from CAD.

Aged ; Case-Control Studies ; Coronary Artery Disease ; genetics ; Female ; Gene Frequency ; Genotype ; Humans ; Male ; Middle Aged ; Nitric Oxide Synthase Type III ; genetics ; Peptidyl-Dipeptidase A ; genetics ; Polymorphism, Single Nucleotide

ObjectiveTo observe the influence of iodine on mRNA expression of iodide transporter (NIS),insulin-like growth factor Ⅰ (IGF- Ⅰ ) and transforming growth factor beta(TGF-β) in thyroid and mammary glands of lactating rats and to explore the role of NIS,IGF- Ⅰ and TGF-β mRNA in iodine uptake in thyroid and mammary glands of lactating rats.MethodsOne hundred and one Wistar rats(80 female and 21 male),weighting 8 - 100 g were selected.These female rats were randomly divided into five groups according to their body weight:control group(NI,normal feed,drank deionized water containing iodine 50 μg/L) ; low iodine group 1 and 2(LI-1,LI-2,low iodine feed,drank deionized water containing iodine 0 and 5 μg/L,respectively); high iodine group 1 and 2(HI-1,HI-2,normal feed,drank deionized water containing iodine 3000 and 10 000 μg/L,respectively),16 rats in each group.After feeding for 3 months,the female and male rates were mated 3:1.The female rats in each group were sacrificed at the fifth and tenth day after postpartum.Thyroid and mammary glands were taken.The mRNA levels of NIS,IGF- Ⅰ and TGF-β in thyroid and mammary glands of lactating rats were determined by real time quantitative PCR.ResultsThe fifth days after postartum,NIS,IGF- Ⅰ and TGF-β mRNA expression levels of thyroid and lactating mammary glands were different between groups,and the differences were statistically significant ( NIS:F =631.46,64.91,all P ＜ 0.01 ; IGF- Ⅰ:F =11.45,6.56,all P ＜ 0.01 ; TGF-β:F =291.83,304.53,all P ＜ 0.01).Compared with control group [NIS:0.0066 ± 0.0023, (0.1481 ± 0.0711 ) × 10-2; IGF- Ⅰ:0.0419 ± 0.0062,0.0542 ± 0.0044; TGF-β:0.1416 ± 0.0277,0.1670 ± 0.0499],regardless of thyroid or mammary gland,the NIS,IGF- Ⅰ and TGF-β mRNA expression of LI-1 [NIS:0.0447 ± 0.0110,(0.3030 ± 0.1831) × 10-2;IGF- Ⅰ:0.0662 ± 0.0078,0.0902 ± 0.008; IGF- Ⅰ:0.5514 ± 0.0508,0.6942 ± 0.0367],LI-2[NIS:0.0317 ±0.0081,(0.3017 ± 0.1601) × 10-2; IGF-I:0.0645 ± 0.0054,0.0894 ± 0.0093; TGF-β:0.5292 ± 0.0332,0.6704 ± 0.0277 ] was significantly increased (all P ＜ 0.01 ); the NIS mRNA expression of HI-1 [0.0043 ± 0.0011,(0.1233 ± 0.0954) × 10-2],HI-2[0.0037 ± 0.0017,(0.1058 ± 0.0854) × 10-2] was decreased(all P ＜ 0.05),while the expression of IGF-Ⅰ mRNA [0.0521 ± 0.0910,0.0715 ± 0.0026; 0.0516 ± 0.0078,0.0697 ± 0.0038] and TGF-β mRNA [0.2087 ± 0.0425,0.2361 ± 0.0425; 0.1971 ± 0.0237,0.2257 ± 0.0752 ] was increased (all P ＜ 0.05 ).The tenth days after postpartum,the mRNA expression levels of NIS,IGF- Ⅰ and TGF-β of thyroid and lactating mammary gland in rats were different between groups,and the differences were statistically significant (NIS:F =103.55,116.32,all P ＜ 0.01; IGF-Ⅰ:F =67.67,11.98,all P ＜ 0.01; TGF-β:F =74.30,381.30,all P ＜0.01 ).Compared with the control group[NIS:0.0069 ± 0.0011,(0.1337 ± 0.0599) × 10-2; IGF-Ⅰ:0.0390 ±0.0071,0.0534 ± 0.0056; TGF-β:0.1351 ± 0.0336,0.1534 ± 0.0320],the mRNA expression levels of NIS,IGF- Ⅰ and TGF-β of LI-1 [ NIS:0.0432 ± 0.0165,(0.2962 ± 0.0985 ) × 10-2; IGF- Ⅰ:0.0643 ± 0.0088,0.0873 ± 0.0055 ; TGF-β:0.5042 ± 0.0912,0.6408 ± 0.0420],LI-2[NIS:0.0287 ± 0.0111,(0.2873 ± 0.0862) × 10-2; IGF- Ⅰ:0.0621 ± 0.0094,0.0862 ± 0.0038; TGF-β:0.4893 ± 0.0504,0.6372 ± 0.0389] were significantly increased(all P ＜ 0.01 ); the NIS mRNA levels of HI-1 [ 0.0042 ± 0.0029,(0.1006 ± 0.0909) × 10-2],HI-2[0.0035 ± 0.0020,(0.0890 ± 0.0119) × 10-2] were decreased(all P＜ 0.05),while the expression of IGF-Ⅰ mRNA[0.0516 ± 0.0078,0.0668 ± 0.0071; 0.0508 ± 0.0089,0.0621 ± 0.0064] and TGF-β mRNA[0.2007 ± 0.0546,0.2175 ± 0.0370;0.1959 ± 0.0393,0.2097 ± 0.0425] were increased(all P ＜ 0.05 ).In thyroid and mammary glands,the comparisons of NIS,IGF,TGF-β mRNA expression of the fifth and tenth day after postartum,between each group were not statistically significant(all P ＜ 0.05).ConclusionsThere are regulatory mechanisms of thyroid and mammary glands of lactating rats in response to low or high iodine conditions.In low iodine,the expressions of NIS,IGF- Ⅰ and TGF-β mRNA in thyroid and mammary glands increase and iodide uptake ability is enhanced to meet the body needs.In high iodine,the expression of NIS mRNA decreases in thyroid and mammary glands.Due to the reduced ability of iodine uptake,iodine intake is reduced,thereby reducing the hazards of high iodine in filial rats.

A BBB co-culture cell model consisting of rat brain microvascular endothelial cells (BMEC) and astrocytes (AS) was established to study the effect of Angelica dahurica coumarins on the transport behavior of puerarin across blood-brain barrier (BBB) in vitro and in vivo. The barrier function of this model was evaluated by measuring the transendothelial resistance, phenol red permeability and BBB related protein expression. The permeability assay and western blot methods were performed to study the effects of Angelica dahurica coumarins on the BBB permeability and the expression of BBB related protein. The animal experiment protocols in this study were approved by the Animal Ethics Committee of Xi'an Jiaotong University (Animal Ethics No.: 2021-1329). The results showed that the established BMEC/AS co-culture model could be used to evaluate drug transport across BBB in vitro. After combined with Angelica dahurica coumarins, the transport capacity of puerarin was significantly increased in vitro and in vivo. Additionally, Angelica dahurica coumarins enhanced BBB permeability and inhibited the protein expression of P-glycoprotein (P-gp), zonula occludens-1 (ZO-1) and occludin. Angelica dahurica coumarins might increase BBB permeability by inhibiting the expression of P-gp and tight junction protein, thereby increasing the content of puerarin in brain tissue.

OBJECTIVETo probe the epidemiological trend of respiratory syncytial virus (RSV) and cellular immunological change of RSV bronchopneumonia among children in Suzhou in the past five years.

METHODS10,205 children with acute respiratory tract infection from January 2001 to December 2005 were enrolled into the study. Nasopharyngeal aspirates were obtained from the respiratory tract by aseptic vacuum aspiration. Direct immuno-fluorescence assay was employed to detect seven kinds of virus antigens including RSV antigen. CD3, CD4, CD8, CD19, CD16 and CD56 in peripheral blood mononuclear cells of 30 patients with RSV bronchopneumonia (1.5-24.0 months old group) were analyzed by flow cytometry analysis, and 15 normal infants (1.5-24.0 months old group) were enrolled as control group.

RESULTSThe annual positive rate of RSV was 24.94%, 25.83%, 24.05%, 25.39% and 27.30% respectively from 2001 to 2005. It also found that the peak season for RSV infection was spring or winter (January to March or November to December). The positive rate of RSV was significantly higher in 1-12 months old group than that in > 12 months old group (chi2 = 97.320, P < 0.01), as well as the groups between 1-12 months old (chi2 = 7.804, P < 0.05, the highest positive rate was occurred at 3-6 months old group). The positive rate of RSV was significantly higher in boys than that in girls (chi2 = 9.693, P < 0.01). The percentages of CD3+, CD4+, CD8+ and NK (CD16 + 56)+ cells were significantly lower in RSV bronchopneumonia than those in control group (t = 3.199, P < 0.01; t = 2.215, P < 0.05; t = 2.619, P < 0.05 and t = 5.240, P < 0.01, respectively). While the percentage of CD19+ cells was significantly elevated in RSV bronchopneumonia than that in control group (t = 2.875, P < 0.01).

CONCLUSIONRSV infection is of obvious seasonal changes. The younger the patient, the higher positive rates of RSV infection is, while and the cellular immunity function is lower. The effective measures for preventing RSV infection are important, especially for the infants. Further investigation is necessary to understand the causes of the variations for RSV infections between boys and girls.

Adolescent ; Bronchopneumonia ; epidemiology ; immunology ; virology ; Child ; Child, Preschool ; China ; epidemiology ; Female ; Humans ; Infant ; Infant, Newborn ; Male ; Respiratory Syncytial Virus Infections ; epidemiology ; immunology ; Respiratory Syncytial Viruses

OBJECTIVETo investigate the interrelations among morphology, immunology, cytogenetics and clinical outcome in childhood acute leukemia with 11q23 abnormalities.

METHODSEighteen patients with 11q23 abnormalities, from 320 childhood acute leukemia patients, were retrospectively analysed for cell morphology, flow cytometry, immunophenotyping, R-banding karyotype as well as clinical features and prognosis. Twenty cases of childhood AL with normal karyotype during the same period were used as control.

RESULTSThe incidence of 11q23 abnormalities in our childhood acute leukemia patients was 5.63% including 14 acute lymphoblastic leukemia (ALL) and 4 acute myeloid leukemia (AML). Of 16 cases immunophenotypically tested, 13 expressed lymphoid antigens and 3 CD(34) and other myeloid antigens. Karyotype analysis disclosed the following abnormalities: t(4; 11)(q21; q23) in 6 cases, t(10; 11)(p13; q23) in 3, t(11; 19)(q23; p13) in one and del(11)(q23) in 6. The complete remission rate for these patients with 11q23 abnormalities was comparable to that of the control (72.2% vs 80.0%, P > 0.05), while the mortality rate in the former was significantly higher than that in the latter (61.1% vs 25.0%, P < 0.05).

CONCLUSIONS11q23 abnormalities were mainly seen in childhood ALL and acute monocytic leukemia with unique prognostic features.

Acute Disease ; Adolescent ; Child ; Child, Preschool ; Chromosome Aberrations ; Chromosomes, Human, Pair 11 ; genetics ; Cytogenetic Analysis ; Female ; Humans ; Immunophenotyping ; Infant ; Leukemia ; drug therapy ; genetics ; immunology ; Male ; Prognosis ; Retrospective Studies

To observe normal bone marrow stromal function for supporting hematopoiesis and realize the correlation of bone marrow stromal function in patients with aplastic anemia (AA) and anisodamine therapy, normal human marrow CFU-GM was assayed on bone marrow stromal layer (SL) formed on week 3 culture, and the CFU-GM without SL as a control (100%). Results showed that the production of CFU-GM on the normal SL was significantly higher than that of the control. The production of CFU-GM on the SL of 4 AA patients, who responded to anisodamine, was < 80% of the control, and when 2 of them were reexamined for stromal function after treatment, the number of CFU-GM on SL rose up to 168.8% and 249.2% from 38.7% and 39.7% of the control respectively. While in the rest 6 patients, the number of CFU-GM was > 80% of the control, only one patient was improved by anisodamine therapy. So, normal bone marrow stromal layer can obviously support the growth of granulocyte/macrophage progenitor cells. Anisodamine therapy could be an effective agents for aplastic anemia with stromal dysfunction
Adolescent ; Adult ; Anemia, Aplastic ; drug therapy ; Anti-Inflammatory Agents, Non-Steroidal ; therapeutic use ; Bone Marrow Cells ; cytology ; Female ; Humans ; Male ; Middle Aged ; Solanaceous Alkaloids ; therapeutic use ; Statistics as Topic ; Stromal Cells ; physiology

OBJECTIVETo explore the feasibility of a new protocol for acute graft versus host disease (aGVHD) prophylaxis in haploidentical bone marrow transplantation.

METHODSThirty-eight high-risk leukemia patients underwent haploidentical G-CSF primed bone marrow transplantation without ex-vivo T cell depletion, the donors were given G-CSF 250 microg/d for 7 days prior to marrow harvest. All patients received a same chemo-radiation conditioning regimen, including cytarabin, cyclophosphamide and total body irradiation (TBI). GVHD prophylaxis regimen consisted of ATG, CsA, MTX, Mycophenolate mofetil (MMF) and anti-CD(25) monoclonal antibody (MoAb).

RESULTSAll patients achieved engraftment of a median of 19 and 22 days for neutrophil and platelet, respectively. Cytogenetic analysis showed 100% donor hematopoietic cells in all recipients after transplantation. One of the twenty patients (5%) experienced grades II - IV acute GVHD. The recovery of CD(3)(+)CD(8)(+) T cells, CD(19)(+) cells and CD(56)(+) cells after transplantation was within 3 approximately 12 months. Of the 20 patients, 16 were alive with minimal and limited chronic GVHD and karnofsky score over 90% in a median follow-up of 9 months. Disease free survival (DFS) rates was (80 +/- 9)%.

CONCLUSIONG-CSF priming marrow graft along with sequential immunosuppressants, especially the addition of anti-CD(25) MoAb for aGVHD prophylaxis could achieve excellent engraftment, proper immune reconstitution and very low incidence of grade II - IV GVHD. The new protocol is effective and feasible in preventing severe acute GVHD and improving DFS.

Adolescent ; Adult ; Antibodies, Monoclonal ; therapeutic use ; Bone Marrow Transplantation ; adverse effects ; methods ; Child ; Female ; Follow-Up Studies ; Graft vs Host Disease ; etiology ; prevention & control ; Hematopoietic Stem Cell Mobilization ; methods ; Humans ; Immunosuppressive Agents ; therapeutic use ; Interleukin-2 Receptor alpha Subunit ; immunology ; Leukemia ; therapy ; Male ; Middle Aged ; Tissue Donors ; Transplantation, Homologous