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.

Objective To investigate the effects of montelukast(MK),a selective cysteinyl leukotriene receptor 1 antagonist on interleukin(IL)-5 and eotaxin expression as well as serum total immunoglobulin E(IgE) production during MK treatment of mouse acute asthma airway inflammation.Methods Sensitized Balb/c mice were challenged by ovabumin(OVA)to establish the acute asthmatic mo-(del).Twenty-five mg/kg of MK(MK group) or Saline(Saline group)were given by intravenously for 3 days.Cellular infiltration of bronchoalveolar larvage fluid(BALF) were assessed by Wright staining.Production of IL-5 and eotaxin in the lung or BALF and serum IgE were determined by enzyme linked immunosorbent assay(ELISA).The expression of IL-5 and eotaxin mRNA were measured by semi-quantified reverse transcriptase-polymerase chain reaction(RT-PCR).Plasma MK level was determined by liquid chromatography.Results After 24 h OVA challenge,the numbers of total white blood cells,neutrophils and eosinophils(EOS)of BALF were(26.0?18.9)?10~7/L,(5.92?8.09)?10~7/L and(0.74?0.88)?10~7/L in MK treatment group.They were significantly reduced compared with those in Saline group,respectively(P80%.The level of IL-5 in BALF and lung tissue were(48.52?14.45) ng/L and(27.40?9.62) ng/g protein in MK group,which significantly declined compared with that in saline group;BALF eotaxin level declined too.Serum IL-5 and total IgE level were also significantly reduced;IL-5 mRNA expression in lung significantly decreased.Eotaxin and its mRNA expression in lung were not decreased significantly.Conclusion MK(exerts) its anti-inflammatory effect mainly through the suppression of IL-5 and IgE production.

Combined with the development of modem hospital pharmacy,in light of the present condition of the continuing education for the hospital pharmacists,based on the continuing education mode and method of the affiliated provincial hospital of Anhui medical university pharmacists,from the establishment of continuing education's goal and mode,the construction of hierarchical continuing education system,and education content integration aspects,this paper expounds the role and influence of the pharmacist professional skill contest.Through pharmacist professional skill contest,pharmacist can test and evaluate the hospital pharmacist professional ability,gradually improve pharmaceutical service level and quality of pharmacists.By introducing the pharmacist professional skill contest,pharmacist can not only timely adjust the continuing education's plan and ways,better coordinate with the development of hospital pharmacists,but also provide important technical support and talent reserves for the pharmacist professional skill contest.

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 investigate the effects of 4-aminosalicylic acid (4-ASA) on rats with 2,4,6-trinitrobenzenesulfonic acid-induced colitis in order to understand its mechanisms in treatment of inflammatory bowel disease (IBD). Methods Thirty SD rats were given 2,4,6-trinitrobenzenesulfonic acid to induce colitis and were divided into model group, 5-ASA (200 mg/kg) treated group and 4-ASA (200 mg/kg) treated group with 10 each. Another 10 rats were severed as normal control. Seven days later,all animals were sacraficed for estimation of colonic tissues. The iNOS and serum level of interleukin (IL)-8 were detected by immunohistochemistry and enzyme-linked immunosorbent assay (ELISA) ,respectively. And the apoptosis of polymorphonuclear neutrophil (PMN) was examined by flow cytometry. Results In comparison with model group, the body weight was increased in rats treated with 4-ASA (t= 14.09,P<0.01), whereas the macroscopic and histological scores and MPO activity were decreased (t=7.87,18.37,6.66 and 19.60,respectively, all P values <0.01), which were similar to 5-ASA treated group (all P values > 0. 05). The expression of tissue iNOS was 73.55%±5.15% in model group, which was higher than that in control group [(5.95±1.45)% ,t=39.93,P<0.01)],but was lower than that in 5-ASA treated group [(37.80±3.82)%,t = 17.62,P<0.01] and 4-ASA treated group [(42.27±3.52) %, t= 15.76 ,P<0.01]. The serum level of IL-8 in model group was significantly higher than that in 5-ASA treated group and 4-ASA treated group (P<0. 01). The apoptosis of PMN in model group was lower than that in control group (t= 11.48,P<0.01), but higher than that in 5-ASA treated group (t= 7.51, P<0.01) and 4-ASA treated group (t= 10.47,P<0.01). Conclusions The efficacy of 4-ASA in treatment of IBD may be related to the mechanisms of reducing the migration and the activities of PMN, up-regulating PMN apoptosis and scavenging reactive oxygen radicals produced by PMN.

Objective To investigate the effects of Gefarnate on expression of myeloperoxidase (MPO),cyelooxygenase-1 (COX-1) and COX-2 in trinitrobenzene sulphonic acid (TNBS) induced experimental colitis in rats and its therapeutic effects on ulcerative colitis. Methods Forty female Sprague-Dawley (SD) rats were randomly divided into 4 groups with 10 each. The rats in group A, B and C were infused with TNBS/alcohol by enema. After the production of colitis, the rats in group A or B were treated daily with 1 ml of normal saline or with 1 ml of 5-ASA (100 mg/kg) by enema,and those in group C were treated daily with 1 ml of Gefarnate by gavage. Group D was served as normal control. After the production of colitis,animals were sacrificed at day 7 and 14 with 5 in each group. The macroscopic changes of the colon were evaluated according to disease activity index (DAD scoring and histological change was assessed by HE staining. MPO activity of the mucosa was detected by biochemical methods. Expressions of COX-1 and COX-2 in tissues were detected by immunohistochemistry. Results Compared with group A, macroscopic and histological scores and MPO activity were significantly decreased in group B and C (P<0.05). The expressions of COX-1 at day 7 and 14 were 1.86±0.51 and 1.96±0.41 in group B, 1.73±0.68 and 1.79±0.6 in group C, 1.91±0.34 and 1.99±0.45 in group D, respectively, which were significantly higher than those in group A (0.87±0.18 and 0.93±0.15, P<0.05). Whereas the expressions of COX-2 at day 7 and 14 were 1.53±0.19 and 0.73±0.15 in group B, 1.73±0.94 and 0.86±0.29 in group C, 0.24±0.18 and 0.18±0. 16 in group D, respectivley, which were significantly lower that those in group A (3.50±0.2;3 and 3.06±0.27). There was a significant difference between group D and group B or C (P<0.05). Conclusions Gefarnate provides a therapeutic effect during TNBS-induced colitis in rats, which is similar to that of 5-ASA. The mechanisms are involved in decreasing the concentration of colonic MPO and regulating the expression of COX-1/COX-2.

Aim To investigate the effect of sulfur dioxide (SO2) on blood pressure and aortic structure of the spontaneous hypertensive rat(SHR).Methods Sixteen male SHRs at the age of 4 weeks were randomly divided into SHR control group and SHR+Na2SO3/NaHSO3 group,respectively.Another eight male WKY rats served as WKY control group.Five weeks later, the blood pressure was determined.The rat aortas were dyed with Hart's Method. The morphometric parameters including outer radius, lumen radius and the wall thickness were calculated by Leica workstation. The proliferative index (PI) of smooth muscle cells was detected by immunohistochemical assay.Results The blood pressure of SHR control increased compared with that of WKY control rats(P

Objective To isolate and culture sweat gland epithelial cells in vitro,and to study the effects of acetylcholine (ACh) on intracellular flee calcium concentration ([Ca~(2+)]i) of cultured sweat gland epithelial cells.Methods Sweat glands epithelial cells were collected by enzymatic digestion.After ACh was added to the primary and first passage cells,[Ca~(2+)]i was examined using confocal laser scanning microscopy (CLSM) and the Ca~(2+) sensitive dye Fura 3/AM.Results The primary and first passage epithe- lial cells grew well.After ACh was added,opening of the calcium channel and significant [Ca~(2+)]i increase were observed when the primary and first passage cells were incubated with high concentration of calcium (2 mmol/L);no significant [Ca~(2+)]i increase was observed in those cultured without calcium.Conclusion Upon stimulation with ACh,calcium channels of cultured primary and first passage sweat gland epithelial cells would open,influx of extracellular Ca~(2+) occurred,which resulted in an increase of [Ca~(2+)]i.Extracellular bound calcium was therefore converted into intracellular free calcium.

OBJECTIVETo investigate immunodominance in CD8+ T cell responses to human immunodeficiency virus type 1 (HIV-1) epitopes.

METHODSFrequency of Interferon-gamma (IFN-gamma) secreting cells and the proliferation percentage of CD8+ T cells in PBMC from an HIV-1-infected long term nonprogressor (LTNP) were assessed after stimulation with either the 34 pools of 701 overlapping peptides covering the regions of HIV-1 Env, Pol, Gag, Vif, Nef, Tat or some single peptides, by using various assays including enzyme-linked immunospot (ELISPOT) and CFSE Carboxy-fluorescein diacetate, succinimidyl ester (CFSE) labeling and flow cytometry.

RESULTSHIV-1 Gag peptides induced the highest frequency of IFN-gamma secreting cells, followed by Nef, Tat, and Vif. Meanwhile, Env and Pol failed to induce significant responses. In the IFN-gamma ELISPOT assay, stimulation with single peptide and the corresponsive peptide pool generated analogous results. In addition, the frequencies of IFN-gamma secreting cells and the proliferation percentage of CD8+ T cells detected-ELISPOT and CFSE labeling and flow cytometry were proportional, when single peptides were used for stimulation.

CONCLUSIONCD8+ T cells can respond to some specific HIV-1 epitopes and induce immunodominant responses. As a complimentary approach to the standard of ELISPOT assay, We recommend a novel CFSE labeling and flow cytometry assay for the examination of immunodominance in studies of HIV-1 specific proliferation percentage of CD8+ T cell responses.

Adult ; CD8-Positive T-Lymphocytes ; drug effects ; immunology ; metabolism ; Enzyme-Linked Immunosorbent Assay ; methods ; Female ; HIV Infections ; immunology ; virology ; HIV-1 ; immunology ; Humans ; Immunodominant Epitopes ; immunology ; pharmacology ; Interferon-gamma ; metabolism

The chemical constituents from Euphorbia lunulata was investigated in this paper. Fourteen compounds were isolated and purified by column chromatographies on silica gel and preparative HPLC. Their structures were identified by physiochemical properties and NMR data analysis as lupeol (1), euphol (2), cassipourol(3) , 24-methylenecycloartan-3beta-ol (4), 24-hydroperoxycycloart-25-en-3beta-ol (5), 25-hydroperoxycycloart-23-en-3beta-ol (6), betulin (7), uvaol (8), (23E) -25-methoxycycloart-23-en-3beta-ol (9), (23E) -cycloart-23,25-dien-3beta-ol (10), 24-methylenecycloartan-3beta, 28-diol (11), salicinolide (12), 2alpha, 3beta, 5alpha, 9alpha, 15beta-pentaacetoxy-11,12-epoxy-7beta, 8alpha-diisobutyryloxyjatropha-6 (17) -en-14-one (13) and 3beta, 5alpha, 15beta-triacetoxy-7beta-isobutyryloxy-9alpha-nicotinoyloxyjatropha-6 (17), 11(E)-dien-14-one (14). Among them, compounds 1-11 were isolated from E. lunulata for the first time.
Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Euphorbia ; chemistry ; Magnetic Resonance Spectroscopy ; Molecular Structure ; Spectrometry, Mass, Electrospray Ionization ; Stereoisomerism