Objective To investigate human immunodeficiency virus type 1 (HIV 1)specific CD8'T cell responses and related human leukocyte antigenⅠ(HLA-I)restriction sites.Methods Four Epstein-Barr virus (EBV)strains-transformed B-lymphoblastoid cell lines (BLCL)from periph eral blood monocytes (PBMC)of 4 healthy individuals were established.Using synthesized peptides in HIV-1 Gag,Vif and Nef regions as epitopes and using BLCL as antigen presenting cells (APC), the frequency of interferon-7(IFN-?)secreting cells in CD8 T cells from a HIV-infected long term nonprogressor by enzyme-linked immunospot (ELISPOT)assay in vitro were assessed.Moreover, matching analysis of HI.A-Ⅰrestriction sites were performed.Results HIV-1 epitopes were presen ted restrictively by HLA-Ⅰmolecules in specific CD8 T cell responses,and this antigen presenting had high specificity.HLA-Ⅰrestriction sitesA03,A26,C04andC08 contributed to presentepitopes in HIV-1 C-ag region,A03 and C08 to HIV-1 Nef as well.Conclusion HLA-Ⅰrestriction sites may include A03,A26,C04 and C08 in HIV-1 Gag region,and A03,C08 in Nef region.

The CrdS protein responding to the acidic adaptation was prokaryotic-expressed in our Laboratory to explore the regulatory mechanism in the acidic adaptation of Helicobacter pylori (H. pylori). The whole genomic DNA of H. pylori strain 26695 was abstracted and set as the template firstly. And then the hp1364 gene coding CrdS protein was amplified via the PCR technique. Then the clonal recombinant plasmid pUCm-T-hp1364 and the prokaryotic expression plasmid pQE30-hp1364 were built and identified by the methods of PCR, cutting with two enzymes and sequencing. After that, the plasmid pQE30-hp1364 was transferred into the E. coli XL1 blue and induced with IPTG. Using western blot and SDS-PAGE, it can be analyzed that the expressed recombinant protein existed mainly in the form of the inclusion bodies and its relative molecular mass was about 46 kDa. The successfully attained recombinant protein CrdS will provide the material to explore the regulatory mechanism in the acidic adaptation of H. pylori and the new way to resist the infection of H. pylori.
Bacterial Proteins ; biosynthesis ; genetics ; Blotting, Western ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; metabolism ; Helicobacter pylori ; genetics ; Plasmids ; Polymerase Chain Reaction ; Recombinant Proteins ; biosynthesis ; genetics

Objective:To evaluate the acute oral toxicity , skin irritation and skin allergy of Thuja essential oil ( TEO) , and pro-vide experimental basis for the clinical use of TEO .Methods:The acute oral toxicity was measured by Horn ’ s assay .Totally 40 KM mice were divided into four groups and intragastrically administered with TEO at different dose of 21.50, 10.00, 4.64 and 2.15 g · kg-1 .After the 14-day observation, the death number and toxic manifestations were recorded and observed , and LD50 was calculated by checking the Horn's form of LD50 .The skin irritation test was performed on healthy adult white rabbits .Totally 9 rabbits were divid-ed into 3 groups randomly , and TEO at the concentration of 100%, 50%and 25%was painted on the skin of the rabbits .Edible vege-table oil was used as the negative control .The erythema and edema of the treated skin were evaluated and scored .Delayed skin hyper-sensitivity reaction was used to investigate the allergy of TEO .Totally 30 white guinea pigs were randomly divided into 3 groups:TEO group, the negative control (edible vegetable oil) and the positive group (1%2, 4-dinitrochlorobenzene).After the intracutaneous in-duction stage and local induction stage , TEO was used to activate the hypersensitive reaction .The skin response was observed and scored after the 24-hour and 48-hour activation.Results:The mice in 21.50 g · kg-1 TEO treatment group were all dead , while only a part of the mice in 10.00 and 4.64 g · kg-1 TEO treatment groups were dead , and no mice died in 2.15 g · kg-1 TEO treatment group.According to the Horn's form of LD50 , LD50 of TEO was 9.26 g · kg -1 for male mice and 7.94 g · kg -1 for female mice.The results of skin irritation test indicated the strong irritation effects of TEO .However , the irritation of TEO was reduced after the dilution , and 25%TEO showed no irritation to the skin of rabbits .The results of delayed skin hypersensitivity reaction showed obvious erythema and edema induced by 2, 4-dinitrochlorobenzene , while no obvious erythema and edema were found in TEO treated guinea pigs , indi-cating non-allergic effect of TEO .Conclusion:TEO has strong skin irritation in rabbits , while no obvious oral toxicity in mice and skin allergy in guinea pigs .

OBJECTIVE: To discuss the optimal operation mode for the management of donated drugs.METHODS: The possible problems existed in the management of donated medicines were found out through summarizing our practical experience in the inventory,storage and use etc of the donated drugs.RESULTS & CONCLUSIONS: In the management of donated medicines,emphasis should be attached to the inventory,storage and use of the donated medicines,meanwhile,a sound supervising system for the donation affairs should be set up to standardize the donation behavior,strengthen expiration date management and gradually establish the practical and effective work mode.

OBJECTIVE:To establish a HPLC method for the determination of ferulic acid in pedo spleen energy activating Misture.METHODS:The determination was performed on Hypersil ODS column,the mobile phase consisted of1%acetic acid-methanol(70∶30)with a flow rate of0.8ml/min,the column temperature was26℃,the detection wavelength was320nm and the sample size was20?l.RESULTS:Good linear relationship with peak area score was achieved when the detection con?centration range of ferulaic acid was within a range of0.02～1.2mg/ml(r=0.9997),the average recovery was97.88%(RSD=1.19%).CONCLUSION:The method can be served as a quality control for pedo spleen energy activating Misture.

Objective:To observe the expression of cell adhesion molecule CD44 in lung tissue of rats with asthma.Methods:32 SD rats were randomly divided into 4 groups:the normal group and the model group,8 rats in each group.The asthmatic model was established by repeated inhalation of ovalbulium.The expression of CD44 in lung tissue of rats was detected by semi-quantitatively reverse transcription-polymerase chain reaction(RT-PCR) and by means of immuno-histochemical staining.Differential leukocytes counts(neutrophils,eosinophils,and lymphocytes) in bronchoalveolar lavage fluid(BALF) were performed after staining with wright staining.Results:The expressions of CD44 increased one week after OVA challenge(P

The present study reported that 5srRNA from reticulocytes of rabbit could pass into the nucleus of mouse myeloma cells SP2/0,and in the mean time DNA synthesis and cells division were markedly suppressed. In a separate experiment, the rRNA was extracted from embryonic liver and erythrocytes of rabbit or rat and analysed by agarose electrophoresis method. The result indicated that the amount of 5srRNA in various period of the development of erythrocyte is changed along with denucleation process. Thus it is likely that 5srRNA of mammalian erythrocyte plays a role in reversing malignant phenotype of tumor cells and promoting denucleation of mammalian erythrocyte through inhibiting DNA synthesis.

ObjectiveTo explore the abilities of automatic processing,controlled processing,selective attention by Stroop color-word test in patients with ischemic cerebrovascular disease.Methods124 patients (aged 60-90 years) with ischemic cerebrovascular disease and 126 cases with age and civilization matched healthy people were examined by Stoop color-word test (SCWT).The SCWT indexes were compared between two groups.Results Reading time of card 1,timeconsuming and error of card 2,4 and Stroop interference effects (SIE) in SCWT had significantly decreased in the patients than in the healthy people (all P＜0.05).ConclusionsThe abilities of controlled processing and selective attention,but not automatic processing are damaged in patients with ischemic cerebrovascular disease.

Objective To evaluate the feasibility of transfection of Sonic hedgehog gene (SHH)into bone marrow mesenchymal stem cells(BMMSC).Methods After the SHH gene was transfected into BMMSC by electroporation apparatus,the transfection rate was evaluated by fluorescence inverted microscope.The growth curves of untransfected and transfected BMMSC were drawn,respectively,to observe the influence of transfection on cells.The expression of SHH gene in the BMMSC was detected by PCR,RT-PCR,Western-blot analyses.Results Through fluorescence inverted microscope,the observed transfection rate was appropriately 30％,PCR showed a obvious increase of SHH expression in transfected cells than that in untransfected cells,and it is quantified by qPCR for appropriately 7 times.Western-blot further demonstrated that the SHH protein expression in transfected cells had a distinct increase.However,it was observed that the exponential phase of BMMSCSHH growth curve delayed.The growth curves of both overlap 12 days after transfection.Conclusions This electroporation method can transfect exogenous SHH gene into BMMSC sufficiently with the effective protein expression in BMMSCSHH.It is the foundation of further research of genetic therapy for ischemic heart disease.

ObjectiveTo investigate the value of CEUS in differential diagnosis of benign and malignant thyroid solid lesions.MethodsTotally 45 patients with thyroid solid lesions underwent CEUS examination.The enhancement mode and blood perfusion characteristics of malignant and benign lesions were analyzed,and the results were compared with pathology.ResultsCEUS diagnosed 31 benign lesions and 14 malignant masses.The diagnostic accordance rate of CEUS was 93.75％ (30/32) and 92.31％ (12/13) for benign and malignant lesions respectively.In 31 benign lesions,27 (27/31,87.10 ％) showed entire homogeneous enhancement,4 (4/31,12.90％) presented inhomogeneous enhancement.In 14 malignant masses,1 (1/14,7.14 ％) showed homogeneous hypoechoic enhancement,12 (12/14,85.71％) presented inhomogeneous hyperechoic enhancement and 1 (1/14,7.14％) presented non-enhancement.ConclusionCombined with two-dimensional grey scale ultrasound and CDFI,CEUS enables dynamic observation of the microvascular perfusion of thyroid lesions,therefore helps to differentiate benign and malignant thyroid lesions.