Administration, Cutaneous ; Adolescent ; Antihypertensive Agents ; administration & dosage ; therapeutic use ; Child ; Clonidine ; administration & dosage ; therapeutic use ; Double-Blind Method ; Female ; Humans ; Male ; Tic Disorders ; drug therapy

Objective To investigate the proliferation and identification of dendritic cells(DC)de- rived from peripheral blood of patients with bladder cancer in vitro.Methods The mononuclear cells were prepared from peripheral blood of patients with bladder cancer by Ficoll-Hypaque centrifugation method,and were induced by the recombinant cytokines hGM-CSF(50 ng/ml),hlL-4(10 ng/ml)and hTNF-?(50 ng/ ml)for 2 weeks.The growth and morphology of DC were observed through the phase contrast or electron mi- croscope,and their pheuotypes were determined by flow cytometry.The capacity of DC to activate T cell-de- pendent anti-tumor immune responses was tested by MTT method.Results The DC cultured in vitro turned into suspensive growth from adhesive situation on the 6th day,then the number of DC increased con- tinuously and the cells showed the irregular morphologic appearance of DC with veiled edges on the 8th day. Flow cytometry showed that the mature DC expressed high levels of specific markers such as CD_(1a),CD_(83), CD_(86)and HLA-DR.T cells activated by DC showed strong cytotoxicity to bladder cancer cell line BIU87 with a killing rate of(48.8?3.7)%,while the killing rate of T cells which were not activated by DC was(25.7?1.5)%;the difference of the rate between them was significant(P＜0.01). Conclusions The DC can be cultured from peripheral blood of patients with bladder cancer by induction of rhGM-CSF,rhIL-4 and hT- NF-?in vitro.This may lay an experimental foundation for further research on DC vaccine.

OBJECTIVE: To investigate a nwe, simple technique for preparation of interferon-alpha-liposomes, which may be suitable for industrial use. METHODS The uniform design coupled with computerized optimization was utilized to screen the formulation and preparation procedure of interferon-alpha-liposomes. Pro-liposomes were prepared by the powder bed grinding method and combined with interferon-alpha-solution to form interferon-alpha-liposomes. Liposome size was determined by the particle size analyzer. Free interferon-alpha and interferon-alpha-liposome were separated by gel filtration. Then the recovered activity of interferon-alpha was analyzed by enzyme-linked immunosorbent assay. RESULTS The result demonstrated that the best interferon-alpha-liposome formulation was as follows: the protectant was sorbitol; weight ratio of protectant to lipid was 5:1; weight ratio of octadecytamin to lipid was 1:9; weight ratio of sobey phosphatidylcholine to cholesterol was 9:1 respectively. Interferon-alpha-liposome size determined by the particle size analyzer was 80.8+/-36 nm and the encapsulation efficiency was 59.0+/-3.3%. CONCLUSION The powder bed grinding method can be used to prepare pro-liposomes which can be easily combined with interferon-alpha-solution to form interferon-alpha-liposomes.

Anticonvulsants ; adverse effects ; Humans ; Seizures ; chemically induced

Purpose To analyze differential diagnosis between Hashimoto thyroiditis of atypical cell clusters ( ACC) and papillary thy-roid carcinoma (PTC). Methods 153 cases of Hashimoto thyroiditis (HT) were collected and divided into 3 groups:PTC group (49 cases), ACC group (32 cases), and HT as control group (72 cases). Morphology observations were done. CD56, CK19, galectin-3 and Ki-67 were detected by immunohistochemistry. Results Morphologic differences were observed among the groups. PTC showed milder positive expression of CK19 and galectin-3, and weaker positive expression of CD56 than that of ACC. Ki-67 index in ACC and PTC was lower than that of HT. Conclusions Morphological characteristics combined with CD56, CK19, Galectin-3 and Ki-67 evalu-ations could be valuable in the differential diagnosis between PTC and ACC.

OBJECTIVETo explore the correlation between Chinese medical (CM) syndrome types of chronic atrophic gastritis (CAG) patients and Helicobacter pylori (Hp) infection, polymorphisms of IL-1B, and IL-1β.

METHODSTotally 192 CAG patients and 202 healthy subjects (as the healthy control group) were recruited in this case-control study. The Hp infection was tested by 13C-urea breath test and colloidal gold-labeled assay (GICA). The concentration of peripheral blood IL-1β was measured by ELISA. The polymorphisms of IL-1B gene in the promoter region were analyzed using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP).

RESULTSPi-Wei weakness syndrome (PWWS) was dominant in CAG patients (31.77%, 61/192 cases). The Hp infection ratio in CAG patients was 53.65% (103/192 cases), of which, Pi-Wei damp-heat syndrome(PWDHS, 64.86%, 24/37 cases) and Gan-Wei disharmony syndrome (GWDS, 66.67%, 24/36 cases) were dominant. Compared with the health control group, the plasma concentration of IL-1β was obviously elevated in CAG patients with PWDHS, GWDS, and static blood obstructing collaterals syndrome (SBOCS) (all P < 0.05). Additionally, there was no difference in the distribution of polymorphisms in the promoter region of IL-1 B gene between the CAG patients and the healthy control group (P > 0.05).

CONCLUSIONSThe incidence risk of CAG was not associated with IL-1B polymorphism. But CM syndrome types of CAG patients was associated with Hp infection and peripheral blood IL-1β levels.

Case-Control Studies ; Gastritis ; Gastritis, Atrophic ; genetics ; Helicobacter Infections ; genetics ; metabolism ; Humans ; Incidence ; Interleukin-1beta ; genetics ; Medicine, Chinese Traditional ; Polymorphism, Genetic

Objective: To investigate the incidence of CMV infection(CMV-I) and CMV related diseases (CMV-D) after allogeneic hematopoietic stem cells transplantation in 70 consecutive allogeneic hematopoietic stem cells transplantation(allo-HSCT) patients and to search for the optimal prophylactic strategy.Methods: Blood samples were monitored using the CMV pp65 antigenemia assay.Of the 70 patients observed,30 patients with chronic myeloid leukemia［CML:CP(27),AP(2),BC(1)］,12 with acute myeloblastic leukemia(AML),10 with acute lymphoblastic leukemia(ALL)and other cases were NHL(3), AA(5), MDS(7), SCLC with pancytopenia (1),CLL(1), and MF (1). Sixty six patients received HLA - identical siblings transplantation and four received tranplants from their HLA- haploidentical donors. Seventy cases included allo-PBPCT (64 cases) , allo-BMT (4 cases) and allo-PB+BMT (2). Before transplantation, all patients and donors received CMV serological examination except 4 pairs of donors/recepients. All 66 patients (3 cases were CMV IgM positive) and 64/66 donors were CMV IgG positive. Results：After transplantation, 64/70 patients developed CMV viremia during monitoring period. Forty three of 70 patients developed CMV-D.Thirty five of them suffered from CMV-associated interstitial pneumonia(CMV-IP). The high peak levels of CMV antigenemia were associated with development of CMV disease　. Close correlation was found between acute graft vs host disease(GVHD) and CMV disease. The patients were followed up for 2 to 24 months. The patients who received preemptive therapy(group A)had significantly better outcome than CMV disease group(group B, P=0.0001). Conclusions: The results suggest that CMV antigenemia has high predictive value for subsequent CMV disease and CMV pp65 antigenemia -guided early therapy has particular advantage for avoiding morbidity and mortality caused by CMV disease.

Three bactreiophages of Pseudomonas aeruginosa were isolated from sewage and named as PaP1, PaP2 and PaP3. All belong to double-strand DNA phages, their genome is about 47kb, 34kb and 24kb respectively. The titre (pfu/mL) of three phages is respectively 109, 1011 and 1011, PaP1 is lytic phage, both PaP2 and PaP3 are lysogenic. Under electron microscope, All show icosahedral heads with diameter of 70nm, 55nm and 65nm respectively. PaPl belongs taxonomically to Myoviridae, and both of PaP2 and PaP3 belong to Pedoviridae. The phage-re-sistance and substitution phenomenon of the resistant flora for the sensitive were observed, and the mutation frequence of Pseudomonas aeruginosa resistant to the phage is about 1.4 ? 10-7 ~ 7.9 ?10-7 determined by end-point -titer method.

As the ubiquitous nucleic acids recognizing motif, Zinc finger protein play important role in regulation of gene expression. The study of recognization specific will greatly facilitate understanding the delicate interaction of Zinc finger protein and DNA. By the choice of expression vector, the induction and culture conditions, the DNA binding domain of Zif268 was expressed in Escherichia coli partly solubly. The gel mobility shift assay shows that purified DNA binding domain can bind its natural target sequence specifically, which indicates the DNA binding domain remains its DNA binding activity in Escherichia coli. The functional expression of DNA binding domain of Zif268 will greatly facilitate the development of in vivo genetic selection assay for the study of Zinc fingers-DNA interaction.
DNA-Binding Proteins ; genetics ; Electrophoretic Mobility Shift Assay ; Escherichia coli ; genetics ; metabolism ; Gene Expression ; Genetic Vectors ; genetics ; Zinc Fingers ; genetics

In order to establish the stable andreliable ISSR-PCR System of Lysimachia christinae, L16 (4(5)) orthogonal design, which based on 7 levels of single factor experiment, were used in this study. The variance analysis was carried out by SPSS 19.0, and 5 main factors affecting the reaction system were optimized in 4 levels. The best annealing temperature was selected by the optimized reaction system. And the stability and reliability of this system was tested by 23 samples from different origins. The results showed that the five factors (DNA template, primer, dNTP, Mg2+ and Taq enzyme) were the most impacts on the amplified results of ISSR-PCR of L. christinae. The order of the influence was: primer > Taq enzyme > DNA template > Mg2+ > dNTP. The optimal system, which was determined by multiple comparison on different levels of each factor, was total volume of 25 microL, including DNA template 60 ng, primer 0.3 micromol x L(-1), dNTP 0.2 mmol x L(-1), Mg2+ 1.8 mmol x L(-1), Taq enzyme 1.25 U. The optimal system was stable and reliable tested by 23 samples from different origins. This study lays the foundation for genetic diversity analysis, fine varieties selection and molecular identification of L. christinae, and provides reference for optimization on ISSR-PCR system of other speciesin future.
DNA Primers ; genetics ; DNA, Plant ; genetics ; Drugs, Chinese Herbal ; chemistry ; classification ; Microsatellite Repeats ; Polymerase Chain Reaction ; methods ; Primulaceae ; classification ; genetics ; Quality Control