Objective To observe the effect of Anshenbunao Syrup on learning and memory capability in mice. Methods The learning and memory capability of mice was investigated by Y-water maze test and step-down test. The contents of protein, DNA and RNA were investigated by Lowry methods, the diphenylamine reagent method and Oricinol reagent method. Result Anshenbunao Syrup could enhance the memory capability of normal mice and improve the memory emersion disorder of mice induced by ethanol. In addition, it could increase the contents of protein and DNA in brain of mice. Conclusion Anshenbunao Syrup could improve the learning and memory capability of normal and memory emersion disorder mice, and its mechanism may be related to promoting protein synthesis in brain.

Objective To investigate whether abnormalities can be detected in normal-appearing white matter(NAWM)and normal-appearing white matter(NAGM)in patients with clinically isolated syndrome(CIS)and comparing them to the abnormalities in relapsing-remitting multiple sclerosis(RRMS)by using diffusion tensor imaging(DTI)histogram.To detect the potential relationship between DTI indices of NAWM,NAGM and patient's clinical condition.Methods Nineteen patients with CIS,19 clinically diagnosed RRMS patients and 19 sex-and age-matched healthy volunteers were included in this study.Conventional MRI and DTI images were obtained using Siemens 1.5 T Magnetom sonata scanner.DTI histograms of NAWM and NAGM were obtained after post-processing.The mean value,peak height,peak location of the histogram were used for analysis.All data was statistically processed with SPSS for Windows.Results NAWM average MD was higher and FA was lower in RRMS[MD(0.83±0.04)×10-3mm2/s,FA 0.36±0.03]when compared to CIS[MD(0.79±0.02)×10-3mm2/s,FA 0.40±0.02]and control[MD(0.78±0.02)×10-3mm2/s,FA 0.41±0.01](P＜0.01).But no statistically significant difference was found between CIS and control.The peak height of NAWM average MD histogram was significantly lower in CIS than control(P＜0.05),while the peak location of average FA histogram shifted to the left(P＜0.01).Patients with CIS[(1.08±0.06)×10-3mm2/s]showed significantly higher NAGM average MD than control[(1.03±0.05)×10-3mm2/s](P＜0.05),but,lower than RRMS[(1.18±0.12)×10-3mm2/s](P＞0.01).There were no correlation between DTI indices and EDSS scores in patients with CIS.Moderate correlation between NAGM average MD(r=0.568,P＜0.05)and EDSS scores were found in patient with RRMS.Conclusion NAWM and NAGM abnormalities do occur in CIS which can be detected by DTI.The underlying pathological changes in NAWM and NAGM in CIS may be milder than RRMS as demonstrated by DTI histogram.

Objective To investigate the effect of highly active antiretroviral therapy (HAART) on human immunodeficiency virus type-1 ( HIV-1 ) antigen specific cytotoxic T lymphocyte (CTL) immune responses. Methods Peripheral blood mononuclear cells (PBMCs) were collected from 38 HIV-1 infected individuals receiving HAART ( HAART group) and 31 HIV-1 infected individuals not receiving HAART (non-HAART group), and stimulated with a peptide pool containing 12 overlapping peptides in HIV-1 P24;then the frequency of interferon γ ( IFNγ ) secreting cells were assessed by enzyme-linked immunospot (ELISPOT) method. Difference in HIV-1 antigen specific CTL immune response between non-HAART group and HAART group was analyzed by χ2 and Mann-Whitney U tests. Results Positive response rate of HIV-1 antigen specific CTL immune responses in HAART group ( 65.8%, 24/38 ) was higher than that of non-HAART group (32.3%, 10/31, χ2 = 6. 522, P ＜ 0.05 ). For HIV-1 infected individuals with blood CD4 +T cells ＞ 350/μL, the frequency of HIV-1 antigen specific CTL responses in HAART group was higher than that in non-HAART group (Z = -2. 819, P ＜0.05 ). In the HAART group, those receiving HAART more than 12 months were of higher frequency of HIV-1 antigen specific CTL responses ( Z =-2. 195, P ＜ 0. 05 ). Conclusion HAART especially long-term treatment may enhance HIV-1 specific CTL responses in HIV-1 infected individuals.

OBJECTIVE: To establish a technical process for the separation and purification of total flavones from Licorice. METHODS: The static absorption capacity of macroreticular adsorptive resins D101、Hz-806、AB-83 for total flavones from licorice were compared. The macroreticular adsorptive resin columns on the Licorice extractives were eluted respectively with different concentrations of ethanol, then the content, the weight of residue and purity coefficient of flavones from Licorice in the eluant were detected. RESULTS: The optimal technological conditions were found in AB-8 as follows: flow rate=3ml/min, sample concentration=1.5 mg/ml, pH=5 and 80% ethanol was used as eluting agent. CONCLUSIONS: AB-8 macroreticular adsorptive resin can effectively separate and purify total flavones from licorice, the purity coefficient thus obtained being over 50%, which meets the requirement of the study of effective components of herbal medicine.

OBJECTIVETo construct dengue virus-specific full-length fully human antibody libraries using mammalian cell surface display technique.

METHODSTotal RNA was extracted from peripheral blood mononuclear cells (PBMCs) from convalescent patients with dengue fever. The reservoirs of the light chain and heavy chain variable regions (LCκ and VH) of the antibody genes were amplified by RT-PCR and inserted into the vector pDGB-HC-TM separately to construct the light chain and heavy chain libraries. The library DNAs were transfected into CHO cells and the expression of full-length fully human antibodies on the surface of CHO cells was analyzed by flow cytometry.

RESULTSUsing 1.2 µg of the total RNA isolated from the PBMCs as the template, the LCκ and VH were amplified and the full-length fully human antibody mammalian display libraries were constructed. The kappa light chain gene library had a size of 1.45×10(4) and the heavy chain gene library had a size of 1.8×10(5). Sequence analysis showed that 8 out of the 10 light chain clones and 7 out of the 10 heavy chain clones randomly picked up from the constructed libraries contained correct open reading frames. FACS analysis demonstrated that all the 15 clones with correct open reading frames expressed full-length antibodies, which could be detected on CHO cell surfaces. After co-transfection of the heavy chain and light chain gene libraries into CHO cells, the expression of full-length antibodies on CHO cell surfaces could be detected by FACS analysis with an expressible diversity of the antibody library reaching 1.46×10(9) [(1.45×10(4)×80%)×(1.8×10(5)×70%)].

CONCLUSIONUsing 1.2 µg of total RNA as template, the LCκ and VH full-length fully human antibody libraries against dengue virus have been successfully constructed with an expressible diversity of 10(9).

Animals ; Antibodies, Viral ; CHO Cells ; Cell Surface Display Techniques ; Cricetinae ; Cricetulus ; Dengue Virus ; immunology ; Gene Library ; Genetic Vectors ; Humans ; Immunoglobulin Heavy Chains ; immunology ; Transfection

OBJECTIVETo delineate the origins of small supernumerary marker chromosomes (sSMCs) identified in 4 infertile males.

METHODSThe sSMCs were analyzed with combined G-banding, N-banding, multiplex ligation-dependent probe amplification (MLPA), fluorescence in situ hybridization (FISH) and single nucleotide polymorphisms array (SNP-array) techniques.

RESULTSG-banding analysis has suggested a 46,X,-Y,+mar karyotype in all of the 4 cases. N-banding revealed that all of the sSMCs have possessed two satellites located on both sides. By MLPA, 1 patient showed copy number gains for 15q11.2 region. SNP-array analysis suggested that all had duplication for 15q11.1-q11.2 region, spanning 3.06 Mb, 0.9118 Mb, 1.728 Mb and 0.287 Mb, respectively. By FISH analysis, all of the sSMCs showed two hybridization signals, indicating that they were dicentric chromosomes.

CONCLUSIONIn all of the four cases, the marker chromosomes have derived from chromosome 15 and were bisatellited and dicentric, which gave rise to a karyotype of 47,XY,+ish,inv dup(15)(q11)(D15Z4++). sSMC 15q11 therefore may be a major cause for male infertility.

Adult ; Chromosome Banding ; Chromosomes, Human, Pair 15 ; genetics ; Female ; Genetic Markers ; Humans ; Infertility, Male ; genetics ; Male ; Pregnancy