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

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.

Methods The maximum tolerated dose( MTD)of water and ethanol extracts of Radix millettiae speciosae were measured for the safety evaluation. Results Maximal tolerated dose( MTD)of water extract was higher than 1 000 g·kg-1 ,which equaled to 110 times the dose for human adults(60 kg). MTD of ethanol extract was higher than 1 700 g·kg-1 ,which equaled to 186 times the dose for human adults(60 kg). Conclusion Radix millettiae speciosae does not have obvious toxicity,thus its routine clinical dose is safe and feasible.

Acquired Hyperostosis Syndrome ; drug therapy ; Adult ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Phytotherapy

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.

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.

Objective To investigate the incidence of thyroid space occupying lesion by ultrasound imaging in patients with breast cancer.Methods From 2009 to 2012,150 patients with breast cancer as experimental group and another 150 patients with breast benign diseases as control group underwent thyroid ultrasound examination.Ultrasound results of the two groups were analyzed to find the differences.Restult More obvious space occupying lesions were found in experimental group than those in control group (P＜0.05).Conclusion The incidence rate of thyroid space occupying lesion in the patients with breast cancer is higher than that in the patients with no breast cancer.

Objective To explore the expressions of smooth muscle actin (SMA) - a and CD44 mRNA in the lungs of rats with asthma in different stages. Methods The asthmatic rat model was established by repeated exposing the animal to aerosolized ovalbumin (OVA). After being sensitized, the rat was subjected to laboratory test. Semi - quantitatively reverse transcription- polymerase chain reaction (RT- PCR) was used to analyze CD44 and a - SMA mRNA of lung tissue. Differential leukocytes(neutrophils, eosinophils, and lymphocytes) counts in bronchoalveolar lavage fluid (BALK) were performed after Wright staining. Results The expressions of CD44 and ? - SMA mRNA increased after OVA challenge 1 week (P