The paper introduces and summarizes the teaching purpose and content project,implementation and effect of the optional course in Excel and medical data analysis.

Objective To establish an aging model of mesenchymal stem cells (MSCs) and to investigate aging related biological mechanism for the purpose of studying the senesence of MSCs .Methods MSCs were separated and purified from human placenta, and the cells of the third passage(P3-MSCs) were cultured in the medium for 2 hours, then 100,200 and 300 μmol/L hydrogen peroxide ( H2 O2 ) was added to the cells for 2 hours to establish the MSCs aging model in vitro. Biological characteristics of aging MSCs were evaluated by cell cycle assay and senescence associated β-galactosidase staining.The expression of p16,p21 and p53 genes was further measured using quantitative real-time PCR (RT-PCR).Re-sults Compared with the control , the number of MSCs treated with 200μmol/L H2 O2 for 2 hours was significantly decreased and the cells displayed less adipogenic ,osteogenic and chondrogenic differentiation .Moreover ,after exposure to 200 μmol/L H2 O2 , the majority of the cells were in the G 0/G1 phase as showed by cell cycle analysis .The percentage of senescence-associated β-galactosidase-positive cells was increased , and the expression of p 16 , p21 and p53 mRNA and protein was significantly increased.Conclusion The results of this study has demonstrated that the H 2 O2 (200 μmol/L) can be used to establish the aging model of MSCs in vitro, and the cellular phenotypic alteration may attribute to the cell cycle associated gene expression (p16, p21, and p53).

We mainly introduce the design of teaching objectives,teaching content,teaching strategies and teaching effect evaluation for the elective course of SPSS and statistical data processing and summarize and reflect on the implementation after teaching combined with the practical teaching of medical statistics.

Objective:To investigate the effect of NEMO binding domain peptide (NBDP) on lung inflammation and apoptosis in mice with acute respiratory distress syndrome (ARDS) and its mechanism.Methods:Thirty-six male BALB/c mice were divided into normal saline (NS) control group, ARDS model group, NBDP negative control group and 6, 12 and 18 μg NBDP pretreatment group by random number table method, with 6 mice in each group. ARDS mouse model was reproduced by aerosol inhalation lipopolysaccharide (LPS) 50 μL. An equivalent among of NS was inhaled in NS control group. The mice in NBDP negative control group were inhaled the materials similar to the non-functional NBDP 30 minutes before the aerosol inhalation LPS; 6, 12 and 18 μg of NBDP 50 μL were respectively inhaled in NBDP pretreatment groups. After inhalation of LPS for 6 hours, mice were sacrificed to get lung tissue and observe the degree of pathological injury and edema. Western blotting was used to detect the phosphorylation of nuclear factor-κB (NF-κB) pathway related proteins [NF-κB inhibitor (IκB) kinaseα/β(IKKα/β), IκBα and NF-κB p65; p-IKKα/β, p-IκBα, p-p65] and the expression of caspase-3 in lung tissue. The bronchoalveolar lavage fluid (BALF) was collected and the levels of inflammatory markers such as myeloperoxidase (MPO), interleukins (IL-1β, IL-8), and tumor necrosis factor-α (TNF-α) were detected by enzyme linked immunosorbent assay (ELISA).Results:ARDS model group had severe edema and hemorrhage, alveolar structure destruction, pulmonary hemorrhage and hyaline membrane formation etc. under light microscope, consistent with the pathological characteristics of ARDS lung tissue, suggesting that the ARDS model was successfully reproduced. ELISA showed that MPO, IL-1β, IL-8 and TNF-α levels of BALF in ARDS model group were obviously higher than those in NS control group. There were no significant differences in the above inflammatory indicators between NBDP negative control group and ARDS model group. The levels of MPO, IL-1β, IL-8 and TNF-α in NBDP pretreatment groups were significantly lower than those in ARDS model group in a dose-dependent manner, especially in 18 μg NBDP, the differences were statistically significant as compared with ARDS model group [MPO (ng/L): 393.32±19.35 vs. 985.87±101.50, IL-1β (ng/L): 43.05±5.11 vs. 97.68±10.88, IL-8 (ng/L): 84.64±2.32 vs. 204.00±17.37, TNF-α (ng/L): 229.13±17.03 vs. 546.73±62.72, all P < 0.05]. Western blotting showed that p-IKKα/β, p-IκBα, p-p65 and caspase-3 protein expressions in ARDS model group were significantly higher than those in NS control group. There was no significant difference in above NF-κB pathway and apoptosis-related protein expression between the NBDP negative control group and ARDS model group. The p-IKKα/β, p-IκBα, p-p65 and caspase-3 protein expression in NBDP pretreatment groups were significantly lower than those in ARDS model group in a dose-dependent manner, especially in 18 μg NBDP, the differences were statistically significant as compared with ARDS model group [p-IKKα/β protein (p-IKKα/β/β-actin): 0.15±0.02 vs. 0.42±0.04, p-IκBα protein (p-IκBα/β-actin): 0.10±0.01 vs. 0.93±0.30, p-p65 protein (p-p65/β-actin): 0.22±0.05 vs. 1.37±0.21, all P < 0.05]. Conclusion:NBDP can inhibit inflammatory response and apoptosis in ARDS lung tissue in a dose-dependent manner, and its mechanism is associated with interference NF-κB signaling pathway transduction.

Objective To compare two sources of mesenchymal stem cells ( MSCs) from human placenta and umbilical cord, and to optimize a technical solution for bench or clinical studies of MSCs.Methods MSCs were isolated from human placenta and umbilical cord and expanded for analysis.The cell morphology was observed under invert microscope, the immunophenotypic feature of MSCs was analyzed with flow cytometer, the cell proliferation ability was determined by cell cycle assay and cell doubling time, the cell differentiation potential was evaluated by osteogenic and adipogenic induction in vitro as well.Results Both sources of MSCs were adherent cells and exhibited fusiform and fibrous morphology. Furthermore, both MSCs high expressed CD90 and CD105, and were negative for the markers of CD34, CD45 and HLA-DR.The population doubling time of MSCs form human placenta and umbilical cord was 39.5 h and 40.8 h separately, and the results of cell cycle analysis showed that the percent of the two sources of MSCs in G0/G1 phase was 52.12%and 57.50% respectively. The above results demonstrated that both sources of MSCs possessed the similar biological characteristics in morphology, phenotype and as well as proliferation ability.In addition, both of them could be induced into osteoblasts and adipocytes in vitro.Conclusion MSCs from human placenta have the similar biological characteristics to these from human umbilical cord, and both of them are better candidates for bench and clinical research.

To observe the cerebral activating effects of needling at Waiguan (SJ5) versus SJ5 plus Yanglingquan (GB34) points in young healthy volunteers based on the hypothesis of "needling effect of combined acupuncture points relates to the brain activation".

ObjectiveTo elucidate the association of immune response to hepatitis B vaccination with HLA-DRB1 * 12 allele as well as the level of IL-4 and IFN-γ.MethodsSeventy-four healthy college students from Guangxi province who had non- or hypor -response to recombinant hepatitis B vaccination and 64 medium- or hyper-responders with the conditions of similar were selected randomly and involved in this study.HLA-DRB1 * 12 was detected by PCR-SSP,the level of IFN-γ and IL-4 cytokines were examined by ELISA.Results(1)The allelic frequencies of HLA-DRB1 * 12 was lower in the non- or hypor-responders than that in the medium- or hyper-responders ( 10.8％ vs 32.8％,P=0.002) ; (2)The expression level of IFN-γ in the non- or hypor-responders ( 7.21±7.92 ) ng/ml was much less than that of the medium- or hyper- responders ( 16.36± 11.00) ng/ml ( P=0.000).(3) The expression level of IL-4 in the non- or hyporresponders (3.18±4.45) ng/ml was much less than that of the medium- or hyper- responders (7.76±5.71 ) ng/ml(P=0.000).(4)No significant differences was seen between the expression level of IFN-γ in the HLA-DRB1 * 12 positive ( 13.18± 11.24) ng/ml and the negative ( 11.00± 10.29 ) ng/ml ( P =0.349 ).(5)No significant differences was seen between the expression level of IL-4 in the HLA-DRB1 * 12 positive (5.947±4.530) ng/ml and the negative (5.132±5.800) ng/ml (P=0.423).ConclusionHLA-DRB1 * 12 might be the allele enhanced immune response to hepatitis B vaccination.The expression levels of IFN-γand IL-4 correlating to Thl/Th2 cells might affect on the immune response to hepatitis B vaccination.

Objective:To investigate the effect of HLA-DRB1*07,13 allelic genes on the immune response to hepatitis B vaccination.Methods:896 healthy college students of Han nationality from Guangxi province,who had received standard courses of vaccination with recombinant hepatitis B virus vaccine,were tested the level of anti-HBs with ELISA method at the sixth month after their last vaccination.The non- or hypo-responders were selected to receive another 20 μg doses of recombinant vaccine and were examined anti-HBs once more four weeks later.99 non- or hypo-responders and 136 medium or hyper-responders were selected for the study subjects.HLA-DRB1*07,13 allelic genes of the subjects were detected by polymerase chain reaction using sequence specific primer (PCR-SSP) method.Results:The frequencies of HLA-DRB1*07 gene in the non- or hypo-responders group were significantly higher than that in the medium or hyper-responder group (16.16% vs 4.41%,P＜0.05). No significant difference of HLA-DRB1*13 gene between the two groups (1.01% vs 3.68%,P=0.389).Conclusion:There is a close relationship between the expression of HLA-DRB1*07 gene and the non- or hypo-response to HB vaccine;No relationship between HLA-DRB1*13 gene expression and the response to HB vaccine is found.

BACKGROUND:Placenta is a valuable source of mesenchymal stem cels for stem cel therapy and future application in the field of regenerative medicine. However, conventional methods cannot acquire a large amount of purified human placenta-derived mesenchymal stem cels. Here, we present a new method for isolating human placenta-derived mesenchymal stem cels suitable for banking strategies and for future clinical applications. OBJECTIVE:To analyze the biological characteristics of human placenta-derived mesenchymal stem cels cultured by tissue dissociating and colagenase digestion. METHODS: Human placenta-derived mesenchymal stem cels were obtained from human placenta by tissue dissociating and colagenase digestion method. Immunophenotype was analyzed by flow cytometry. Growth curve was determined by MTT assay, and differentiation ability was evaluated byin vitro adipogenic, osteogenic and chondrogenic induction as wel. RESULTS AND CONCLUSION:Human placenta-derived mesenchymal stem cels could be passaged stablyin vitro. Furthermore, the cels expressed CD73, CD90, CD105, but were negative for the markers of CD11b, CD19, CD34, CD45, and HLA-DR. Human placenta-derived mesenchymal stem cels proliferated actively and began to grow logarithmicaly at days 3-5 folowed by a plateau period at day 6. In addition, the isolated cels could be induced into adipocytes, osteocytes, chondrocytesin vitro. In a word, the results of this study demonstrated that the tissue dissociating and colagenase digestion method is an efficient method for obtaining a large amount of human placenta-derived mesenchymal stem cels that can be stably cultured in vitro and have strong proliferative ability.

AIM:To investigate the expression of extracellular matrix metalloproteinase inducer(EMMPRIN)and hepatocyte growth factor(HGF)in non-small-cell lung carcinoma(NSCLC)and their relationship with lymphoid metastasis and prognosis.METHODS:Expression of EMMPRIN and HGF in 77 cases of patients with NSCLC was detected immunohistochemically.The relationship of expression of EMMPRIN and HGF with tumor size,smoking,histological type,differentiation,lymphoid metastasis,clinical stage,and prognosis was analyzed.RESULTS:The expressive rates of EMMPRIN and HGF were 68% and 44%,respectively.The expressions of EMMPRIN and HGF were associated positively with lymphoid metastasis(r=0.371 and 0.339,P0.05).The expression of EMMPRIN was associated with the expression of HGF in NSCLC.CONCLUSION:The expression of EMMPRIN and HGF is associated with lymphoid metastasis and prognosis in NSCLC.Overexpression of EMMPRIN and HGF implies infavourable prognosis in NSCLC.