Objective:Got high purity dendritic cells from PBMC recovered from one time used no leukocytes transfusion apparatus.Methods:Rinsed PBMC with natural salt water from one time used no leukocytes transfusion apparatus and induced PBMC with GM-CSF and IL-4.Results:Got 5.1?10 7 PBMC from every one time used no leukocytes transfusion apparatus and 4.7?10 6 DCs with a purity of 96.9% were obtained.These DCs express CD86(96.1%)?CD40(88.9%)?CD83(91.6%)?HLA-DR(92.9%)?HLA-ABC(99%)?CD54(97.5%) highly and a few number of these DCs could stimulate proliferation of allogeneic mixed leukocyte reactions strongly.Conclusion:The results of this research supplied basic data for further research of DC and its clinical application.

This article aims to study the present condition and disadvantages of current obstetrics and gynecology teaching and discuss the urgent needs and methods of evidence-based medicine (EBM) introduction to it.

Objective To observe the effects of ankle intensive training on the recovery of lower limbs motor in stroke patients in later stage of rehabilitation. Methods 30 stroke patients with course ≥ 3 months were divided into the treatment group (n=15) and control group (n=15). The control group received routine treatment, and the treatment group received ankle intensive training in addition. They were assessed with Fugl-Meyer Assessment (FMA) of lower extremity and 10 m walking speed. Results The scores of FMA and 10 m walking speed improved in both groups after treatment (P<0.05), and improved more in the treatment group than in the control group (P<0.05). Conclusion Ankle intensive training can promote the motor recovery of lower limbs in later stroke patients

Objective: To observe the effect of Tuina and respirometric training on the pulmonary function, dyspnea, motor ability, and quality of life in patients with chronic obstructive pulmonary disease (COPD) in remission stage. Method: Sixty-six cases of COPD in remission stage were divided into Tuina group (n = 33) and control group (n = 33) with table of randomized number. After 3-month treatment, the grade of dyspnea and quality of life, pulmonary function, and 6-min Self Walk Test were compared with that before treatment. Results: The pulmonary function of patients in Tuina group were improved remarkably, the improvement in the control group was poor. The qualities of life were enhanced in the two groups, and Tuina method was better than the respirometric training.Both the treatment methods could increase the walking distance in 6 min Self Walk Test and improve dyspnea, without a difference in the two groups. Conclusion: The two treatment methods had good therapeutic effect on dyspnea, quality, motor ability and pulmonary function in patients with COPD in remission stage, and especially, Tuina treatment had a better effect on pulmonary function and quality of life than the control method.

Purpose　To introduce a method to isolate cDNA clones using bacteriophage P1-derived artificial chromosome (PAC) or bacterial artificial chromosome (BAC) as probe for hybridization and try to find some novel genes related to hepatocellular carcinoma.　Methods　PAC 579 (D17S926 locus) and BAC 1529 (D17S1529 locus) in the deletion region of chromosome 17p13.3 in human hepatocellular carcinoma were chosen to screening the human liver cDNA library as probe for hybridization. The isolated positive cDNA clones were partially sequenced, then analyzed by computer comparison and Southern blot.　Results　After three cycles of screening, 78 and 8 candidate positive cDNA clones were isolated using PAC 579 and BAC 1529 probes respectively. Further analysis indicated 18 cDNA clones isolated by PAC 579 probe and 5 cDNA isolated by BAC 1529 probe were potential novel genes related to hepatocellular carcinoma.　Conclusions　The isolation of cDNA clones using PAC and BAC probes is effective and practical.

Objective To determine the effect of Mycobacterium tuberculosis spp. inducing transformation of THP-1 cells to epithelioid cells (EC) and the involved signaling pathways and their regulations. Methods THP-1 cells infection models respectively infected with M. tuberculosis strains H37Rv, bovis and phlei were established. Indirect immunofluorescent staining assays were used to detect the expressions of monocyte/macrophage differentiation antigen CD115 and EC differentiation antigen CD82 of the THP-1 cells before or after infection. By Sandwich ELISA Kits, the phosphorylation levels of p38MAPK, Akt1 and STAT3 of the THP-1 cells before or after infection were measured. The alterations of CD115 and CD82 expression levels were examined when the associated signaling pathways were blocked with specific blocking agents. Results CD115 expression was weakened and CD82 expression was strongly increased in all the THP-1 cells infected with the three strains. A temporal up-regulation of the p38MAPK phoshporylation level but no obvious alteration of Akt and STAT3 phosphorylation levels after THP-1 cells infected by strain H37Rv or bovis. The THP-1 cells infected with anyone of the three strains continuously expressed CD115 after MAPK, PI3K/Akt or JAK/STAT of the cells was blocked. Although JAK/STAT was blocked, the THP-1 cells respectively infected with the three different strains still expressed CD82. However, CD82 expressed in THP-1 cells infected by the strain H37Rv or bovis was disappeared when p38MAPK and PI3K/Akt pathways of the cells were blocked. Conclusion Strain H37Rv and bovis can induce the infected THP-1 cells transforming to EC and p38MAPK and PI3K/Akt signaling pathways participate and regulate the transformation procedure. Of the two signaling pathways p38MAPK seems to be more important.

Objective To investigate the effects of age and gender on anti-oxidative,antiinflammatory and anti-apoptosis functions of high-density lipoprotein (HDL).Methods Totally 120 healthy subjects aged from 20 to 60 years were randomly divided into young and middle-aged male (n=60) group and female (n =60) group,and the 120 healthy elderly aged from 60 to 78 years divided into elderly male (n =60) and elderly female (n =60) groups.Serum levels of high-and low-density lipoprotein cholesterol (HDL-C,LDL-C),total cholesterol (TC),and triacylglycerol (TG) were detected.Content of malonaldehyde (MDA) was detected to determine anti-oxidative function of HDL.Adhesion assay of endothelial cells and monocytes (THP1) was adopted to test the protective effects of HDL on endothelial cells.The expressions of endothelial cell adhesion molecules,VCAM-1 and ICAM-1,were analyzed by Western blot.MTT and flow cytometry assays were used to detect the viability and apoptosis of the cells to test anti-apoptosis function of HDL.Results The levels of low-density lipoprotein,triglycerides and total cholesterol were higher in elder female group than in other three groups (all P＜0.05).The level of HDL-C was higher in young and middle-aged females than in other three groups(all P＜0.05).The level of MDA was higher in elder female group than in other three groups(all P＜0.05).The level of MDA was higher in elder male group than in the young and middle-aged male and female groups(all P＜0.05).After protection of HDL,the number of monocytes adhesion and expression levels of VCAM-1 and ICAM-1 were higher in elder groups than in young and middle-aged male and female groups(all P＜ 0.05).Relative survival and viability rates of endothelial cells were higher in young and middle-aged groups than in elder groups (all P＜0.05).Conclusions Ageing in both male and female induces impairments of anti-oxidative,anti-inflammatory and anti-apoptosis functions of HDL,with more evident decrease in anti-oxidative function in females.

Objective To analyze mRNA expression of silent information regulator 6 (SIRT6) gene in the blood of population with family history of longevity in Bama county of Guangxi and to explore its association with SIRT6 gene polymorphism and its protein Methods One hundred and thirty-seven people (aged 30 ~ 106, 70 males, 67 females, 6.57% Han nationality, 93.43% Zhuang and Yao nationalities) with family history of longevity (long-lived family history group), and 91 people (aged 22~89, 51 males, 40 females, all Zhuang and Yao nationalities) without family history of longevity were recruited in the study (non-long-lived family history group). Real-time fluorescence quantitative RT-PCR was used to detect mRNA expression of SIRT6 gene in two groups. Results SIRT6 mRNA expression of total and femals in long-lived family history group were higher than those in non-long-lived family history group (P<0.05). mRNA expression of GG and CG genotype of SIRT6 (rs350846) and G allele carriers in long-lived family history group were all higher than those in non-long-lived family history group and the difference was statistically significant (P<0.05). SIRT6 mRNA expression was not associated with serum SIRT6 protein in long-lived family history group (P>0.05). Conclusion The expression of mRNA in SIRT6 gene may be associated with familial aggregation of longevity in Bama County of Guangxi and high expression of mRNA of SIRT6 in G allele carriers contributes to longevity.

Objective To construct lentiviral vectors containing peptide P1-GFP fusion genes.Umbilical cord derived mesenchymal stem cells were infected with lentivirus carrying peptide P1 and GFP fusion genes.To inject the targeted umbilical cord derived mesenchymal stem cells into mice and to detect GFP expression in the spleen.Methods Umbilical cord derived mesenchymal stem cells were cultured with adhered tissues of umbilical cord smaller than 1 mm3 . Lentiviral vector containing P1-GFP fusion genes with engineering technology was constructed and infected the umbilical cord derive mesenchymal stem cells.Targeted umbilical cord derived mesenchymal stem cells were intravenously injected in the mouse tail vein and after 18 hours GFP expression was detected with immunohistochamical staining of the spleen tissues.Results Harvested umbilical cord derived mesenchymal stem cells grew well in culture medium. Green fluorescence on umbilical cord derived mesenchymal stem cells were observed under fluorescence microscope at 18 hours after infected with lentivirus.Green fluorescence intensity of umbilical cord derived mesenchymal stem cells was increasing over time and reached a peak at 72 hours.Umbilical cord derived mesenchymal stem cells highly expressed CD105 (90.0%)/CD44 (98%) and CD73 (85.0%)/CD90 (98.5%) molecules.GFP expression was detected in the spleen after intravenous injection of targeted umbilical cord derived mesenchymal stem cells in the mice 18 hours later.GFP expressing cells intimately contacted with lymphocytes.Conclusions Targeted umbilical cord derived mesenchymal stem cells contain P1-GFP fusion genes are constructed.Targeted umbilical cord derived mesenchymal stem cells can be targeted to mouse spleen and intimately contact with lymphocytes after intravenous injection.Our results lay the groundwork for further studies.

OBJECTIVETo study the immune responses of lymphocytes after activated by dendritic cells (DCs) loaded with cytotoxicity T lymphocyte (CTL) epitope based peptide of human alpha-fetoprotein (hAFP, 218-226 LLNQHACAV).

METHODSGet high purity DCs by cultured plastic-adherent monocytes isolated from healthy donor of HLA-A2(+) peripheral blood with granulocyte-monocyte colony-stimulating factor (GM-CSF) and interleukin-4 (IL-4) for 7 days. Stimulate self-lymphocytes with DCs that loaded with CTL epitope based peptide of hAFP under the culture medium contains interleukin-2 (IL-2) for 7 days. Analyse IL-12 and TNF in culture medium and also the specific lysis activity of lymphocytes against four strains of primary hepatocellular carcinoma cells.

RESULTSAfter stimulated by DC loaded with CTL epitope based peptide derived from hAFP, lymphocytes appeared a good characteristics and the culture medium of activated lymphocytes contained a high level Th1 type cytokines of IL-12 and TNF. Activated lymphocytes not only specifically lysed HLA-A2(+) HepG2 line but also had the cytotoxicity against other three primary hepatocellular carcinoma cell lines and T2 target cell loaded with peptide of hAFP.

CONCLUSIONSThe results of this research supply the basic materials for the DC based vaccine with HLA-A2 restricted peptide epitope derived from hAFP against AFP positive primary hepatocellular carcinoma.

Adult ; Animals ; Dendritic Cells ; immunology ; Epitopes ; HLA-A2 Antigen ; immunology ; Humans ; K562 Cells ; Mice ; Peptides ; immunology ; T-Lymphocytes, Cytotoxic ; immunology ; Tumor Cells, Cultured ; alpha-Fetoproteins ; immunology