Mesenchymal stem cells(MSCs) have became the hot spot in cell tissue engineering,cell replacement therapy,gene therapy and transplant research fields. Recent studies have shown that changes in oxygen concentrations affect many biological characteristics of MSCs. Under different oxygen concentrations,MSCs have different proliferation,differentiation,migration and chemotoxis abilities. Hypoxia is a kind of common pathophysiological status,which can promote the proliferation,apoptosis,migration and chemotoxis abilities of MSCs,while hypoxia impacts the differentiation ability depending on different cell types. The mechanism of these response might be involved in hypoxia-inducible factor-1(HIF-1) ,chemokines and their receptors,and matrix metalproteinases. Hypoxia can activate HIF-1 signaling pathway,which upgrades the expression of stromal-derived factor-1(SDF-1) ,and forms microenvironments which stem cells are adapted to and grow in. SDF-1 increases the adhesion,migration and homing of circulating CXCR4-positive progenitor cells to ischemic tissue,and promotes degradation of extracellular matrix,then enhances the migration ability of MSCs by modulating the expression of matrix metalloproteinase and its protein as well.

To explore the teaching model and effect of emergency care simulator(ECS) teaching in "Shock"during clinical bed side teaching,fifty-six students were randomly divided into two groups, receiving model and traditional teaching model respectively. After one semester,the students’examination results and general effect were evaluated. The students’ex-amination results,clinical theory and clinical skills in ECS teaching group were significantly better than traditional teaching group.

Objective To study the influence of sole massage on children’s analepsia from general anesthesia.Methods One hundred children in analepsia period were divided into the test group(50 cases)and the control group(50 cases).The routine nursing and monitoring during recovery stage were done in the control group.Based on the routine monitoring and nursing care as in the control group,the test group received sole massage for 10 minutes.The two groups were compared in terms of declining percentage of SpO2, rate of nausea,blood pressure,heart-rate variability,and post-analepsia dysphoria scores.Result The test group was significantly lower than the control one in all the indexes of the declining percentage of SpO2,rate of nausea,blood pressure,heart-rate variability,and post-analepsia dysphoria scores(all P<0.05).Conclusions Sole massage during children’s analepsia from general anesthesia is effective not only in lessening their declining percentage of SpO2,rate of nausea,blood pressure,heart-rate variability, and recovery dysphoria mark,but also easing their dysphoria,anxiety and fear.It may create a good condition for children to live through analepsia period successfully and safely.

Objective To compare the therapeutic effect of conventional treatment and conventional treatment plus external application of Shuangbai Powder for patients with wounded limb injured by venomous snake. Methods One hundred patients bitten by venomous snake were randomized into treatment group and control group, 50 cases in each group. The control group was given conventional treatment including repeatedly washing the wound with hydrogen peroxide, debriding the wound, letting blood and draining toxicity, local blocking with chymotrypsin, wet packing the wounded limb with magnesium sulfate, and injection with anti-venomous serum, tetanus antitoxin, antibiotics, furosemide and energy mixture. The treatment group was given external application of Shuangbai Powder on the basis of the treatment for the control group. Both groups were given the comprehensive nursing of psychological nursing, wound nursing, dietary nursing, defecation nursing and functional nursing. The swelling-subsiding time for the wounded limb and pain scores of visual analogue scale (VAS) in the two groups were compared. Results(1) After treatment, swelling-subsiding time for the wounded limb of the treatment group was shorter than that of the control group, the difference being statistically significant (P < 0.05). (2) After treatment for 4 days and at the end of the treatment, VAS scores of the two groups were decreased compared with those before treatment(P < 0.05), and the decrease of VAS scores in the treatment group after treatment for 4 days was superior to that in the control group (P < 0.05). Conclusion The conventional treatment plus external application of Shuangbai Powder is an effective therapy for patients with wounded limb injured by venomous snake by shortening swelling-subsiding time and relieving pain for the wounded limb .

BACKGROUND: CpG oligodeoxynucleotide (ODN) is a type of highly effective immune adjuvant with low toxicity, which has an extensive application in gene therapy for many diseases. However, the specificity for species and cells leading to low uptake by cells and degradation by nuclease blocks its clinical application. OBJECTIVE: To explore the specific delivery and its immunologic efficacy of CpG ODN targeting B lymphocytes of umbilical cord blood by CD40 ligand-receptor-mediated carrier system. DESIGN, TIME AND SETTING: An observation and control experiment was performed at the Department of Hematology, and Department of Pediatric, the First Affiliated Hospital of Jinan University from April 2004 to October 2007. MATERIALS: Fresh umbilical cord blood with heparin was obtained from healthy, natal infant. Informed consent was obtained from his parents, and the experiment was approved by the hospital Ethics Committee. METHODS: CD40 ligand (CD40L)-EDC-PLL-CpG ODN conjugated complex was prepared. Mononuclear cells (MNCs) from umbilical cord blood were co-cultured with conjugated complexes. Uptake rate, mean fluorescence intensity of FAM marked CpG ODN, expressions of MNCs, proliferations of lymphocytes and the IgG levels of culture supematants were detected by flow cytometry, fluorescence techniques, MTT assay and ELISA, respectively. MAIN OUTCOME MEASURES: The uptake rate, the mean fluorescence intensity of CpG ODN by MNCs, subgroups and proliferations of lymphocytes, and IgG levels of culture supematants. RESULTS: Compared to the pure CpG ODN group, the uptake rate of the conjugated complexes group was higher (98%), the peak level of up-taking occurred earlier, and intracellular fluorescence intensity maintained much more stable. Expressions of CD19+, CD22+, and CD20+ was increased, A value and IgG levels in supematants were all higher than that of the control group. CONCLUSION: CD40 tigand-receptor-mediated carrier system is helpful for CpG ODN delivery targeting to B lymphocyte, enhancing its immunological efficiency.

98%), the peak level of uptake occurred earlier, intracellular fluorescence intensity maintained much more stable. Expressions of CD19+, CD22+, CD20+ increased significantly. A_~570 values of MNCs proliferation and IgG levels in supernatant were all higher. CONCLUSION: CD40 ligand-PLL carrier system may delivery CpG ODN targeting to B lymphocytes, enhancing its immunological efficiency.

Objective To compare the proteome difference between multiple myeloma cell line U266 cells treated and untreated with PS-341, to investigate the potential drug targets, and to provide theoretical evidence for clinical therapy of multiple myeloma. Methods Two-dimensional gel electrophoresis (2-DE) was performed to separate proteins from treated and untreated U266 cells with proteasome inhibitor PS-341. ImageMaster 2D Platinum software was used to analyze 2-DE image, and matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS) was used to identify the differentially expressed proteins. The expression levels of differential protein BAG-2 in the 2 groups of U266 cells lines were detected by Western blot. Results The 2-DE reference pattern of treated and untreated U266 cells with PS-341 was established. A total of 31 differential proteins were identified by MALDI-TOF-MS, 27 of which were down-regulated after PS-341 treatment. The differential expression level of BAG-2 in the 2 groups of U266 cells was confirmed by Western blot. Conclusion Some down-regulated proteins may be the potential drug targets of proteasome inhibitor PS-341.

Objective To investigate the expressions of TLR2 and TLR4 in patients with psoriasis vulgaris and the efrect of methotrexate(MTX)on them.so as to explore the therapeutic mechanism of MTX in psoriasis vulgafis.Methods Forty-three patients with psoriasis vulgaris were recruited into the study together with 30 normal human controls.Oral MTX was given to patients with an interval of 12 hours for three times per week until the control of conditions followed by 4 weeks of mainmining treatment.The dosage of MTX was 5 mg initially and decreased to 2.5 mg in the maintaining period.Flow cytometry was used to detect the expression of TLR2 and TLR4 in peripheral blood CD14~+ cells from the controls and patients at baseline,4 and 8 weeks after the beginning of treatment.Results The expression rate of TLR2 and TLR4 in CD14~+ cells was(92.6±4.3)%and(48.5±4.6)%,respectively,in untreated patients,significantly higher than that in normal controls(botll P＜0.01).A significant increase was observed in the expression rate of TLR2 and TLR4 in patients with active psoriasis compared with those with inactive psoriasis [(97.5±4.1)%vs(87.6±5.6)%,(55.3±5.8)%vs(40.7±7.1)%,both P＜0.05].Eigh weeks after the beginning of treatment with MTX.the expression rate of TLR2 and TLR4 significantly decreased to (79.6±6.7)%and(34.6±5.9)%.respectively(both P＜0.05).The psoriasis area and severity index(PASI)score had no significant correlation with the expression rate of TLR2 or TLR4(r=0.24.0.27,both P＞0.05).Conclusions TLR2,TLR4 and innate immune response mediated by both receptors play an important role in the pathogenesis of psoriasis.MTX may exert its therapeutic effect on psoriasis by inhibiting the expression of TLR2 and TLR4.

AIM: To design,prepare and screen out functional small interfering RNA(siRNA) for specifically silencing proliferating cell nuclear antigen(PCNA) gene expression and effectively inhibiting cell proliferation on human hepatoma cell line SMMC-7721,human gastric carcinoma cell line SGC-7901 and human colorectal carcinoma cell line Caco2.METHODS: PCNA siRNA was designed based on previous studies about design guidelines and synthesized in vitro by T7 Mega short script reaction kit according to the manufacturer's instructions.After purification,the integrities of siRNA were identified through 19% denaturing polyacrylamide gel electrophoresis,and the concentrations of the generated siRNA were measured by testing the absorbance at 260 nm using a spectrophotometer.Four synthesized double-strand siRNA were transfected into three types of carcinoma cell lines by LipofectamineTM2000 reagent,respectively.WST-8 assay was employed to examine the proliferative inhibitions of these three cell lines.The subsequent alterations on PCNA mRNA and protein levels were determined by semi-quantitative reverse transcription-polymerase chain reaction(RT-PCR) and immunocytochemistry,respectively.RESULTS: 3 sequences,No.2,No.3 and No.4 PCNA siRNA showed effective inhibitions on tumor cells among the 4 candidate siRNA,and a single dose of 50 nmol/L of No.2,No.4 PCNA siRNA showed the most effective inhibitory rates as more than 62% at 48 h after transfection.50 nmol/L of No.2 and No.4 PCNA siRNA transfection caused 72% decrease of PCNA mRNA level and almost completely loss of the protein in human Caco2 cells.CONCLUSION: No.2 and No.4 PCNA siRNA have been screened out in this study,which show the capability to effectively down-regulated PCNA expression and significantly inhibit the proliferation of carcinoma cells.The optimal concentration is 50 nmol/L and satisfactory effects are achieved 48 h after transfection in vitro.

AIM: To evaluate the effect and safety of Rhodamine 123 (Rh123)-mediated photodynamic treatment (PDT) on acute graft versus host disease. METHODS: An acute graft versus host disease (aGVHD) mice model was established using C57B/6 mice as donors and BALB/c mice as recipients. Mixed lymphocytic cells were cultured with Rh123 (50 nmol/L) and irradiated by argon laser 30 mW/cm2 for 3 min, then transplanted to BALB/c recipient mice mixed with donor bone marrow. Hepatopoietic recovery, aGVHD occurrence, survival time after transplantation and pathological changes were observed. In addition, CD3+CD69+ positive rates of MLC were examined by flowcytometry. RESULTS: Occurrence of aGVHD decreased, degree of pathological manifestation became milder, survival rates were higher than non PDT groups. CD3+CD69+ rates of MLC cells treated with photodynamic therapy (PDT) and cultured for 24 h significantly decreased. CONCLUSION: Rh123-mediated PDT can effectively prevent aGVHD of allogeneic bone marrow transplantation in mice.