Objective To study the correlation between the mutation in platelet activating factor (PAF) acetylhydrolase gene mutation(Val279 Phe)and cerebral vascular disease (CVD)in Chinese.Methods Genomic DNA was analysed for the mutation allele in 153 patients with cerebral vascular disease and 100 healthy subjects matched for age and sex without CVD by a specific polymerase-chain reaction. Plasma PAF acetylhydrolase activity was determined by the method of Stafforini et al.Results The prevalence of the mutation and the frequency of the mutation allele are significantly higher in patient with CVD than those in control subjects( P

The forecast of health human resources demand is the most important and one of the most complex aspects of health manpower planning projections. There are many methods in forecasting human resources demand,and each method has its own advantages and disadvantages. Five kinds of commonly used basic methods are comparatively analysised in this paper.

Our aim in this study was to determine differentially expressed genes of CD34 + cell from bone marrow and G CSF mobilized peripheral blood. CD34 + cells isolated from bone marrow and G CSF mobilized peripheral blood of the same donor were identified for differentially expressed genes in CD34 + cells using the technique of suppression subtractive hybridization. Eleven genes were expressed with higher levels in CD34 + cells from mobilized peripheral blood, as compared with data of GenBank. These genes included nuclear proteins, transcriptional regulatory molecules, zinc finger proteins and interferon induced molecules. These results demonstrate that there are some differences between the two groups of CD34 + cells. Further study would give us more extensive understanding about mobilization and homing of hematopoietic stem cells.

Objective To observe the in vivo gene expression profile of the recombinant adenoassociated-2 virus mediated human GM-CSF, mouse GM-CSF (rAAV-2-hGM-CSF, rAAV-2-mGM-CSE )vector modified bone marrow mesenchymal stem cell (BMSC). Methods We transduced the BMSC by rAAV-2-hGM-CSF, rAAV-2-mGM-CSF at the condition which have acquired before respectively, then transfused the in vitro gene modified BMSC after 12 days proliferation in vitro to 6 weeks old nude mice through tail vein,while the BMSC transfused in control group hadn' t been gene modified. 2, 4, 6, 8 weeks after transfusion, count the total white blood cells and detect the hGM-CSF, mGM-CSF concentration in nude mice serum at that time point. Results Nude mice serum hGM-CSF levels were 23.77, 25.32, 19.77, 15.25 ng/L at 2, 4, 6, 8 weeks after transfusion compare to 36.25 ng/L, the in vitro level before transfusion; mGM-CSF levels were 34.96, 34.84, 35.50, 32.93 ng/L at 2, 4, 6, 8 weeks after transfusion compare to 25.14 ng/L, the in vitro level before transfusion; at the same time point the nude mice serum mGM-CSF levels were 17.34,17.44, 14.68, 16.85 ng/L in control group, rAAV-2-mGM-CSF transduced BMSC made the nude mice white blood cell count increased, but no changes in nude mice white blood cell count at rAAV-2-hGM-CSFtransduced BMSC and control group. Conclusion BMSC as a gene therapy vehicle, it can be gene modified in vitro, then the gene modified BMSC could let the therapeutic gene to have therapeutic effects in vivo.

Objective To investigate the differences and harmonization of immunoassay systems in detecting CA19-9 and to assess the possibility of mutual recognition in different laboratories.Methods Data were collected and analyzed from External Quality Assessments (EQA) of NCCL(Lots:200811-201215) and ZJCCL(Lots:080309-120211).120 fresh serum with different concentrations of CA19-9 were collected.The CA19-9 results of healthy people were also collected from September 2010 to March 2012.Four kinds of stable immunoassay systems were involved in our research,including Abbott Architect i2000,Beckman UniCel DxI 800,Roche E170 and Siemens ADVIA Centaur XP.The differences among four system groups were calculated with the EQA data.The fresh serum comparisons were also performed following the guideline of CLSI EP9-A2 The 95％ confidence interval of each immunoassay system was calculated.Comparisons were made by scatter diagrams and weighted regression.Results Both EQA of NCCL and ZJCCL showed better correlation coefficients and larger bias (bw ranged from 1.340 to 4.683) than in fresh serum comparisons.Although the correlation coefficients were all unsatisfactory,the bw were all close to 1 in fresh serum comparisons.When the recommended serum concentration of 27 U/ml was used,the biases were Abbott-Roche-6.41％,Beckman-Roche-5.07％,Siemens-Roche 13.15％,Beckman-Abbott 2.46％,Siemens-Abbott 22.52％ and Siemens-Beckman 39.66％,respectively.Differences of 95％ confidence intervals were statistically significant among parts of the immunoassay systems.Conclusions Only in the lower concentration can CA19-9 results be mutual recognized among four different immunoassay systems,there will be larger differences and risks in the higher concentration.

Objective: To investigate the expression of MDR1 and GFP in the human hematopoietic cells mediated by adeno-associated virus. Methods: The GFP gene was transferred into the human hematopoietic cells by AAV vectors and created strong visible fluorescence by purely molecular biological means. Using adeno-associated virus vectors, we have transferred human mdr-1 gene into human hematopoietic cells and investigated the drug resistence of human hematopoietic cells modified with mdr-1 gene. PCR analysis confirmed that mdrl cDNA had been successfully transferred into the human hematopoietic cells. An assay of MTT proved that the human hematopoietic cells modified by mdrl gene had resistance to colchicine. Results: It was about 30% of the hematopoietic cells that expressed the green fluorescent proteins. The resistance of hematopoietic cells was increased parently when the cells were infected by the crude virus stocks. Conclusion: It is conducted that the AAV vector could successfully transfer the foreign gene into the human hematopoietic cells. The cells modified with mdrl gene have increased the resistance to drugs.

Objective To approach effect of Yiqihuoxue decoction on remnant stomach paralysis syndrome after subtotal gastrectomy. Methods A total of 44 patient of remnant stomach paralysis syndrome were randomly recruited into a control group and a treatment group, with 22 patients in each group. The control group was treated with TPN parenteral alimentation. On this basis, the treatment group was treated with Yiqihuoxue decoction and mental intervention. The clinical effects were observed in both groups after the treatment. Results The total effective rate was 54.55% and 100% in the control group and the treatment group respectively, showing statistic difference(P=0.00625 ＜0.01).Conclusion It was effective to treat remnant stomach paralysis syndrome with Yiqihuoxue decoction, TPN parenteral nutrition and mental intervention.

Objective To investigate the sensitivity of BIOMED-2 primer system in adult acute lymphoblastic leukemia (AIJ,) patients Ig gene rearrangement, and to analyze their frequency, corearrangement pattern, utilization of V, D and J genes and composition of junctional regions. Methods Amplification of rearranged IgH (complete and incomplete), IgK, IgK-Kde and IgL was performed in standard PCR in 29 adult ALL patients. Monoclonal PCR products were subjected directly to DNA sequencing. Sequences were identified by comparison with all known human Ig germline sequences to analyze the recombination patterns, somatic mutations and germline gene segments usage. Results IgH, incomplete IgH, IgK, igK-Kde and Igl, rearrangements were found with positive rate of 70.8%, 12.5% , 29.2% , 25.0% and 0 of B-ALL patients, respectively. All B-ALL patients displayed at least one pattern of Ig gene rearrangements. In TALL, one of five patients was found with incomplete IgH rearrangement, two patients were found with IgK rearrangements and two patients were PCR-negative. The sequence analysis showed that the most frequently used V, D, J segments in adult B-ALL patients were from VH3/VH4 families, DH3 family and JH6 family, respectively. Four of five IgK rearrangement used VκI family. 23.5% B-ALL IgH contained scattered replacement mutations with replacement to silent substitution ratio < 1 in complementarity determining regions. Conclusion BIOMED-2 multiplex PCR analysis strategy is a reliable and useful technique in the adult BALL patients.

~3H-Clonidine, a potent and selective alpha2-adrenergic agonist was used to label alpha2-a-drenoceptors in intact lymphocytes isolated from rat spleen. Binding of ~3H-Clo-nidine was rapid(t1/2: 2min)and readily reversed by 10umol?L~(-1) clonidine(t1/2: 3-4min). ~3H-Clonidine saturationexperiments indicated a single c1ass of site with a K_D of 6.57?1.63nM and Bmax of 72.4?13.4 fmol/10~7 lymphocytes. Adrenergic agonists competed for ~3H-clonidine binding site with anorder of potency:epinephrine)norepinephrine)isoproterenol. These results show the presence ofalpha2-adrenoceptors in splenic lymphocytes. Computer analysis of competition experiments withadrenergic agonists revealed three classes of sites: high affinity site, medium affinity site and lowaffinity site. The affinity of high affinity site is 2-3 orders of magnitude higher than the one ofmedium affinity site, whereas the latter is the same orders higher than low affinity site Using im-proved Mishell-Dutton method, 10umol?L~(-1) clonidine suppressed the Ig M synthesis and thesuppression was blocked by 10 umol?L~(-1) phentolamine. These results indicate the suppression ofIg M antibody response to SRBC in vitro by clonidine is mediated by the alpha-adrenoceptors.

Objective:To clone and express human Fas activation domain(FasAD)with the bioactivity.Methods:The FasAD cDNA was amplified by seminested RT PCR,and then inserted into the prokaryote vector of pTYB12 expressed intein protein to construct the recombinant plasmid of FasAD pTYB12.The FasAD peptide was expressed and purified in IMPACT TM CN system by the method of one step affinity purification.Results:Sequence analysis revealed that cloned FasAD cDNA sequence was completely same as that of Genebank record(M67454).Soluble fusion protein was successfully expressed by induction of IPTG.The FasAD peptide with a molecular weight of 5 000 was obtained by purification and was recognized by the rabbit anti human Fas polyclonal antibody in Western blot analysis.It’s activity of inhibition of apoptosis induced by rhFasL can each to 70% in primary biological detection.Conclusion:The above results indicated that FasAD peptide could be prepared by using the IMPACT TM CN system,thus laid a relatively experimental foundation for further research of relationship between structure, function and interaction with it’s ligands,and for further development of biological immune modulator. [