Objective To evaluate the relationship between proportion of PR1 specific cytotoxic T lymphocytes (CTLs) in the peripheral blood and prognosis and curative effect in patients with HLA-A0201 positive chronic myelogenous leukemia (CML),and to discuss whether PR1 peptide could be used as the following immune therapeutic method for patients who had achieved the standard of stop treatment.Methods The soluble HLA-A0201/PR1 tetramer and flow cytometry were applied to determine the proportion and the frequency of PR1 specific CTLs in peripheral blood from 28 HLA-A0201 positive CML patients.The proportions were compared among different phases of patients.The correlations between the proportion of PR1 specific CTLs and clinical parameters were analyzed.Results There was a negative correlation between PR1 specific CTLs and PCR (bcr-abl/abl)Is (r =-0.658,P ＜ 0.001).The frequencies of PR1 specific CTLs at 3-month,6-month,9-month,12-month,2-year,3-year,4-year,5-year,6-year were (0.06±0.02) ％,(0.10± 0.02) ％,(0.14±0.02) ％,(0.16±0.02) ％,(0.20±0.03) ％,(0.18±0.03) ％,(0.18±0.01) ％,(0.17±0.05) ％ and (0.18±0.03) ％,respectively.The frequency of PR1 specific CTLs at 3-month,6-month or 9-month was statistically different compared with that of the other time spots (P ＜ 0.05),and there were no statistical differencies among the frequencies at 1-year,2-year,3-year,4-year,5-year,6-year (P ＞ 0.05).For patients treated with IM 400 mg qd,the frequency of PR1 specific CTLs in high-risk group was lower than that in low-risk or intermediate-risk groups.Conclusion PR1 specific CTLs can be detected in patients who achieved good curative effect,and is correlated with tumor burden,which indicates that PR1 specific CTLs may be related to the action of resisting leukemia and provide the evidence for PR1 peptide as a potential immune therapeutic schedule in patients who have achieved stable MR45 and MR50.

Objective To investigate the effect of dendritic cells(DC)pulsed with tumor lysate antigens of K562/A02 co-cultured with cytokine induced killer(CIK)cells on cytotoxicity against multidrug resistant(MDR)leukemia K562/A02 cells with high expression of p-glycoprotein(P-gp).Methods Bone marrow mononuclear cells isolated from healthy adult donors were induced to obtain CIK cells and DC respectively and then CIK were co-cultured with DCs which were pulsed with tumor lysate.The morphologic features were observed with optical microscopy and expressions of markers typical were measured using flow cytometry.The cytotoxicity of three effect cells was measured with MTT assay respectively.Results After pulsing DCs with tumor lysate antigens co-cultured CIK or DC-CIK led to a significant increase of lymphocyte proliferation of both populations.The cytotoxicity of CIK co-culturing with DCs pulsed with tumor lysate antigens to K562/A02 and K562 cells was(42.90?0.67)%、(49.85?0.28)%(63.36?0.46)% and(23.56 ?0.43)%、(26.11?0.34)%、(34.46?0.35)% respectively at an effector-to-target ratio of(5∶1、 10∶1 、20∶1).Conclusion This study indicates that DC pulsed with tumor lysate antigens co-cultured with CIK can improve lymphocyte proliferation and enhance the cytotoxic activity.Especially,it can induce a P-gp specific anti-leukemic immune.

Objective To study the role of cutaneous nerve and protease activated receptor 2 (PAR2)in the development of pruritus in atopic dermatitis(AD).Methods Dermal sheets were prepared from chronically pruritic skin lesions of 7 patients with AD,as well as from the normal skin of 7 healthy human controls.Double labeled immunofluorescence was performed using mouse anti-protein gene product 9.5(PGP9.5)monoclonal antibody and rabbit anti-substance P(SP)polyclonal antibody to observe the morphological changes in cutaneous nerve fibers,and Image-Pro Plus 6 software was used to semiquantitively assess the length,diameter of nerve fibers,integral optimal density of PAR2 and SP in dermal sheets.Results Immunofluoresence double staining showed that PAR2 co-expressed with PGP9.5 or SP in cutaneous nerve fibers.Compared with the normal control skin,both the total length and average diameter of PGP 9.5-expressing nerve fibers were increased(11051.8±1900.9 μm vs 7264.0±2659.9 μm,4.23±0.15 μm vs 3.95±0.15 μm,both P＜0.01)in pruritic lesions,while only the average diameter of SP-expressing nerve fibers was up-regulated(3.99±0.20 μm vs 3.80±0.07 μm,P＜0.05),and the total length of them remained unchanged(4304.7±1455.0 μm vs 3380.0±1735.4 μm,P＞0.05).Also,increased integral optimal density was observed for SP and PAR in pruritic lesions in comparison with the normal control skin (27.71±16.52 vs 12.63±4.31.35.99±8.63 vs 22.69±9.56.both P＜0.05).Conclusion Our results indicate a hyper-plasia of cutaneous nerve fibers in chronic itchv skin lesions of AD and an increase in the expression of PAR2 and SP in the cutaneous nerve fibers,suggesting that the signal enhancement in PAR2 pathway may be related to the mechanism of pruritus in patients with AD.

AIM:To determine Radix et Rhizoma Rhici in Ganning Granula(Radix et Rhizoma Rhei,Rhizoma Chuanxiong). METHODS: In order to determine five anthraquinone aglycones of Radix et Rhizoma Rhei,the HPLC system consisted of Nucleosil ODS column,methand-0.1% phosphoric acid(85∶15) mixture as mobile phase,with detection wavelength at 430 nm. RESULTS: HPLC showed that the linearity ranges were as follows: aloe-emodin concentration was at the range of 0.041 6-0.291 2 ?g,the rhein concentration was at the range of 0.037 1-0.259 8 ?g,the emodin concentration was at the range of 0.045 8-0.320 3 ?g,the chrysophanol concentration was at the range of 0.049 60.347 2 ?g,the physcion concentration was at the range of 0.020 6-0.144 5 ?g.The average recoveries of five anthraquinone aglycones were between 96.6%-98.4%.CONCLUSION: The method is simple,accurate,sensible and with good repeatability and can be used for quality control of Ganning Granule.

Objective To evaluate the effect of exogenous wild PTEN gene stable transfected into human ovarian cancer cell line HO-8910 on phosphatidyl inositol 3-kinase( PI3K)/protein kinase B (Akt)siganal pathway and cells proliferation. Methods Wild-type PTEN recombinant eukaryotic expression plasmid was constructed and then was transfected into HO-8910 cells by lipofectamine 2000. The expression of PTEN, Akt1, Akt2, PI3K mRNA and protein of PTEN were tested by reverse transcription( RT)-PCR and Western blot. The proliferation of HO-8910 after wild PTEN gene transfected was measured by methyl thiazolyl tetrazolium(MTT). Results Wild-type PTEN gene was successfully transfected into HO-8910 cells. The results of RT-PCR and western bolt showed that there were the significant expression high level of PTEN mRNA and protein after infected by wild-PTEN plasmid than those in the control[ ( 17 372 ±23)vs.(39±1 )vs. (78 ±4)copies/ml,P ＜0. 05 ]. While the expression of mRNA of Akt1, Akt2 and PI3K were decreased clearly than those in the control [ (28 ± 2 ) vs. ( 115 ± 5 ), (7 ± 1 ) vs. ( 18 ± 2), (61 ± 2 ) vs.(84 ± 2)copies/ml , all P ＜ 0. 05 ]. The proliferation rate of HO-8910 cells was obviously slower than those in the control (90 158 ±47 vs. 148 251 ±65 vs. 250 115 ±62, P＜0.05). Conclusion Transfection of PTEN may increase the expression of PTEN and inhibit the proliferation of HO-8910 cells, in which PI3K/ Akt siganal pathway is inhibit significantly.

Objective To study the correlation between psychological state and coping styles of primary caregivers of stroke patients.Method 286 primary caregivers were screened out by the random sampling method for the investigation of their psychological states and coping styles using SCL-90 and the medical coping modes questionnaire(MCMQ).Results There were significant differences between primary caregivers and Chinese norm in terms of all factors(P<0.01).The major caregivers with higher education were scored significantly lower than those with less education on the factors of depression,anxiety,hostility,fear,psychoticism (P<0.01).There were significant different differences between primary caregivers and Chinese norm in coping style(P<0.01): the score on confrontation in the coping style lower than the norm while the score on avoidance in the coping style higher than in norm.The confrontation and avoidance in the primary caregivers were negatively correlated to the factors of SCL-90(P<0.05)and yielding in the coping style positively to the factors of SCL-90(P<0.05).Conclusions The primary caregivers of stroke patients are at the poor psychological states and they tend to take the passive and negative coping styles.The more they take active coping style,the better their mental states grow and vise versa.Therefore,the caregivers need professional training so that they can use the supports from their families and be encouraged to speak out their poor moods.Post-discharge visits to them,besides,are helpful for the improved psychological states for the improvement of their quality of life.

Objective To investigate the awareness,willingness,motivation,and influencing factors of outpatients for participating drug clinical trials,and provide references for decision-making of drug clinical trials.Methods An amnonymous survey was conducted in the departments of internal medicine,surgery,gynecology,and obstetrics of a randomly selected tertiary referral center,and the results were statistically analyzed.Results A total of 1 067 available questionnaires were received.The total awareness rate of clinical trials was 31.02％,which was closely correlated with age and the degree of education.40.86％ of respondents were willing to participate in drug clinical trials.And 55.28％ of them chose yes because of the willingness to contribute to the development of medical science.People having cognition on clinical trials had more willingness to participate in drug(OR:1.361,95 ％ CI:1.042-1.777).59.14％ of the respondents refused to participate in drug clinical trials,68.62％ of whom refusing to participate mainly worried about the safety of drugs.57.37％ of the respondents comfirmed that they might change their idea if experts were involved.41.33％ were willing to accept training about clinical trials.Conclusion Investigators'overall cognition on clinical trials is closely correlated with the willingness to participate in drug clinical trials.There should propagandize drug clinical trials to make sure the improvement of drug clinical trial progress.

The epidemic characteristics and genotype of Bacillus anthracis strains in Liaoning Province ,China was analyze in this study .Six Bacillus anthracis strains from 2001 to 2011 were studied with multiple‐locus variable‐number tandem repeat analysis (MLVA) .BioNumerics4 .0 software was used to analyze the DNA fingerprint of statistics ,and cluster analysis results were obtained .Clustering analysis found that the 6 strains could be divided into two genotypes .For anthrax outbreaks ,the ge‐netic markers of multiple‐locus variable‐number tandem repeat were highly similar .It's suggested that MLVA is quite useful for investigation of strain relatedness in regions of outbreaks .

To explore the impact of Salmonella on Enterococcusfaecalis isolate N41 under the coexisting,the growth traits and the transcription of 26 virulence genes of N41 at various growth phases were detected.Salmonella and/or N41 were inoculated and done plate counting,then growth curves were drawn and bacterial total RNA were extracted at given time points,quantitive realtime PCR was used to analyze the RNA transcription levels of 26 virulence genes of N41.The result showed that comparison with single incubation,the bacteria concentration of N41 dropped about 3.9 times,and Salmonella dropped about 110-times.Among 26 virulence genes of N41,the RNA transcription levels of 12 virulence genes such as ebpA,ebpC,rnjB,ace,ebpR,psr and so on were promoted at the four growth phases,but the RNA transcription levels of SlyA and sprE were dropped.Except that the RNA transcription levels of CylL-S,CylL-L,efaA and AS had no significant changes at the four growth phases,the mostly rest genes increased dramatically at post-log phase.This indicated that when incubated together,N41 inhibited the growth of Salmonella significantly,and Salmonella promoted the transcription levels of virulence genes of N41 as a whole.The results lay a foundation for further research on the interaction between bacteria species and the pathogenicity mechanism of Enterococci.

Objective To investigate the expression levels of the CD14 mRNA and TLR4 mRNA in peripheral blood mononuclear cells from patients with Parkinson disease and to explore the clinical signicance. Methods For?ty-four patients with Parkinson disease and 37 healthy controls were recruited. We recorded age of onset, duration of ill?ness and sex of all recruited patients. PD patients were evaulated using the Hoehn-Yahr stages, Unified Parkinson Dis?ease Rating Scale (UPDRS) Ⅱand UPDRS Ⅲ,non-motor symptoms scale (NMSS) on“off”time. Reverse transcrip?tion-polymerase chain reaction was performed to determine the expression levels of CD14 mRNA and toll-like receptor 4 (TLR4) mRNA. Results The expression levels of CD14 mRNA(1.459±0.658)2-△△CT and TLR4 mRNA (1.408±0.698)2-△△CT was significantly up-regulated (P<0.05) in Parkinson disease compared with controls((1.162 ± 0.631)2-△△CT、(1.122 ± 0.557)2-△△CT). In addition, there was positive correlation between the expression levels of CD14 mRNA in Parkinson dis?ease patients with the Hoehn-Yahr stages. Meanwhile, there was no significant correlation between the expression levels of CD14 mRNA and TLR4 mRNA with other clinical scores. Conclusions There is positive correlation between the ex?pression levels of CD14 mRNA in Parkinson disease patients with the Hoehn-Yahr stages,indicating that CD14/TLR4 positive monocyte may be involved in the pathogenesis of the Parkinson disease.