AIM To explore the effect of tacrolimus on maturity and allostimulatory activity of cultured dendritic cells (DC) in vitro. METHODS Twenty rats were randomly divided into 4 groups: control, tacrolimus, LPS, and tacrolimus+LPS groups. The bone marrow cells of rats were cultured with granulocyte-macrophage colony-stimulating factors and interleukin-4 (IL-4) for 6 d and the DC which adhered to the wall were harvested. No other extraneous reagents were used in control group. Tacrolimus 10 μg·L-1 was added to tacrolimus group at the beginning of culture. Lipopolysaccarides (LPS) 100 μg·L-1 were administered 18 h beforeharvest in LPS group. In tacrolimus+LPS group, tacrolimus and LPS were used in accordance with tacrolimus and LPS groups. The immunophenotypes of DC were analyzed with flow cytometry and the level of IL-12 secreted by DC was detected by ELISA. The allostimulatory activity of DC on allogeneic T cells was assessed with mixed lymphocyte reaction. RESULTS Compared with control group, LPS increased CD80 and CD86 expressions, IL-12 secretion and allostimulatory activities of DC. Compared with control and LPS groups, tacrolimus cut down the expressions of CD80 and CD86, decreased the secretion of IL-12 and reduced the allostimulatory activity of cultured DC. CONCLUSION Tacrolimus exerts a negative effect on the maturation and allostimulatory activity of cultured DC in vitro.

BACKGROUND: Immune tolerance is regarded as the most effective measure to overcome rejection. For the past few years, the important effect of immature dendritic cells (imDCs) on immune tolerance has drawn close attention. OBJECTIVE: To study the effect of imDCs treated with tacrolimus (FK506) on imlnune tolerance in rat allograft organ transplantation and to investigate the mechanism of immune tolerance induced by imDCs. DESIGN: A randomized and controlled animal experiment. MATERIALS: The experiment was carried out at the Experimental Animal Center, the Affiliated Hospital of Medical College. Qingdao University from April 2006 to December 2006.Cervical heart transplantations were performed in which 45Wister rats were used as donors and45 SD rats as allograft recipients. The ratswere randomly divided into three groups with 15 for each. METHODS: Normal sodium, imDes. and imDCs treated with FK506 were injected via vena caudalis seven. days before the operations respectively. Mixed lymphocyte reaction (MLR) was meas ured on SD, Wistar, and Lewis rats. MAIN OUTCOME MEASURES: Heart allografi survival was monitored and one-way MLR, heart pathology and the levels of interleukin-2(IL-2),IL-4,IL-10,and interferon-Y(IFN-Y)in the serum were detected. RESULTS: In treatment group without FK506.heart allografl survival were prolonged after imDC injection(P＜0.01);while their survival were prolonged further in treatment group with FK506(P＜0.05).MLR showed that the tolerance was donor specific. Analysis of variance showed that tere was high significant difference for serum concentrations of IL-2,IFN-Y,IL-4,and IL-10 in the three groups(P＜0.01).The concentrations of IL-2 and IFN-Y expressing on Th1 were lower. and that of IL-4 and IL-10 expressing on Th2 were obviously higher in the latter two groups. CONCLUSION: ImDCs can induce immune tolerance in rat heterotopic heart transplantation successfully; ImDCs treated with FK506 can enhance the tolerance which is donor specific.ImDCs may induce the immnune tolerance bymeans of modifying the immune response type of T cells (immune deflection from Th1 to Th2), mediating the generation of regulatory T cells (T-reg)and inducing T cells disenabling.

OBJECTIVE To observe the effect of voriconazole on pulmonary fungals infection after renal transplantation.METHODS Nine cases of pulmanory fungals infection after renal transplantation were analyzed to evaluate the therapeutic effect of voriconazole.RESULTS Treated with voriconazole,seven in nine patients were cured,while the other two died.CONCLUSIONS For patients with pulmanory fungals infection after renal transplantation,the application of voriconazole can receive satisfactory effect.

OBJECTIVE:To establish an HPLC method for the determination of Stillbene glycoside (THPG) in Zhipingk-ang misture.METHODS:The chromatographic column was Hypersil C18(250 mm?4.6 mm,5 ?m).The mobile phase consisted of acetonitribl-water(25∶75) at a flow rate of 1.0 mL?min-1 with detection wavelength set at 320 nm. RESULTS: There was a good linear relationship within the concentration range of 0.5～8.0 ?g for THPG. The average recovery of THPG was 100.03%(RSD=1.07, n=6).CONCLUSION: The adoptd method is accurate, reproducible and suitable for the quality con-trol of Zhipingkang misture.

Objective:To investigate the relationship between levels of plasma thrombopoietin(TPO) and thrombocytopenia.Methods:Sixty-eight thrombocytopenia with different causes were injected with rhIL-11 domestically made, by 25 ?g/(kg?d) for 10 days continuously. The plasma TPO levels were determined by quantitative sandwich enzyme immunoassay technique in thrombocytopenic patients between before and after treatment.Results:(1)The serum TPO controls of patients with thrombocytopenia after acute leukemia(AL) chemotherapy were lower than normal controls. The plasma TPO levels in aplastic anemia(AA) and myelodysplastic syndrome(MDS) patients were all elevated compared to those in normal controls.The serum TPO levels in patients with idiopathic thrombocytopenic purpura(ITP) and liver cirrhosis had no difference with the normals. Bone marrow megakaryocytes(MK) count in after chemotherapy AL and AA patients were lower than those with normal serum TPO levels controls. (2)In all above patients used interleukin-11(rhIL-11), their TPO levels were close to normal in efficiency patients. On the contrary, their TPO did not change dramatic. Those react to rhIL-11, their TPO levels were inversely correlated with platelet count.Conclusion:Testing serum TPO levels would be helpful for distinguish the thrombocytopenia with different causes, and provides theorical support to patients to use rhIL-11 reasonably.

OBJECTIVE:To evaluate the drug use status of Chinese materia medica injection(CMMI)in our hospital.METHODS:Chinese materia medica injection(CMMI)used in our hospital from2002to2004was analyzed statistically in terms of consumption sum,defined daily dose(DDDs)and the average daily consumption sum(DDDc).RESULTS:The consumption sum of CMMI in3years increased year by year with annual increase rates of29.7%and32.3%,respectively.The cardio-cerebral vascular CMMI steadily dominated the first place in terms of consumption sum and DDDs in3years with a steady increase year by year from a high starting point.The DDDc of antitumor CMMI ranked the toppest.As for the consumption sum for single variety,xuesaitiong steadily ranked the first place in3years.CONCLUSION:The fast increase of the consumption of CMMI shows that it has a great market demand.

Objective To explore the organic micro environmental effect of skeletal muscles on the proliferation of thoracic malignant cells, its significance in the rarity of metastases in skeletal muscles and the prospect for its clinical applications.Methods Primary culture of new born Wistar rat skeletal muscle cells was established.The murine skeletal muscle conditioned medium(MMCM)was prepared to test its effect on thoracic malignant cell lines of A549、Anip-973,PLA-801C,NCI-H466,Eca109 and benign cell line of BHK-21 by MTT assay.Results Proliferations of thoracic malignant cell lines of A549,Anip-973,PLA-801C,NCI-H466,and Eca109 were significantly restrained when cultured with MMCM,while the proliferation of benign renal cell line(BHK-21)was not affected.Conclusions The conditioned medium of new born Wistar rat skeletal muscle cells could selectively inhibit the proliferation of thoracic malignant cells in vitro.Moreover,it affects tumor cells only and has no apparent effect on normal cells,which differs from most of the chemotherapeutic agents.These findings suggest a sound mechanism in the rarity of metastases in skeletal muscles.A therapeutic agent could be generated from MMCM to complement surgery and/or chemotherapy.

Objective To establish a bedside available risk scoring system of no-reflow in the acute stage of STEMI.Methods Data from STEMI patients treated with PCI divided into model group and validation group were analyzed.Multivariable binary logistic regression analysis was used to identify independent no-reflow predictors of the model group.Finally,a score according to the odds ratio on logistic regression analysis was designed,and then risk stratification was established,and no-reflow high-risk patients with myocardial infarction were selected.The authenticity and reliability of the logistic regression courses were validated using receiver operator characteristic curve (ROC)and Hosmer-Lemeshow goodness-of-fit.Results Multivariate logistic regression analysis demonstrated that female (OR =0.587,P =0.019),Killip class of myocardial infarction≥2 (OR =3.656,P <0.01),TIMI flow ≤2 before primary PCI (OR =0.774,P =0.013),thrombus burden score ≥4 on baseline angiography (OR =2.629,P <0.01),pain to balloon time ≥ 6 h (OR =1.485,P =0.083)were independent correlate predictors of no-reflow phenomenon in the STEMI after PCI.The risk score system demonstrated a good risk prediction in the model group with AUC of 0.716 (95%CI:0.671 -0.761)based on ROC analysis.There was no significant discrepancy between multivariate logistic regression analysis and Hosmer-Lemeshow goodness-of-fit (χ2 =1.027,P =0.994).In risk stratification,total value <2 was assigned into low risk level,and 2-5 was put into the medium risk level,and >5 was arranged into high risk level.The risk score system demonstrated a good risk prediction in the validation group with AUC of 0.891 (95%CI:0.822 -0.959)based on ROC analysis.ROC analysis in the validation group was applied to Killip class,thrombus burden,score and risk stratification in the validation group ,and the no-reflow score was more accurate,with a larger area under the curve (AUC = 0.851,95% CI:0.776 -0.927 ).Conclusions Establishment of no-reflow scoring system with STEMI patients undergoing PCI was benefit to select high risk patients with no-reflow.

Objective By comparing the change of expression of AQP1 in vitro lung preserved by the continuous infusion of a heart‐lung machine ,continuous pressure perfusion and single low temperature ,to explore the best method of vitro lung preserva‐tion .Methods Thirty Mongrel dogs were randomly divided into 3 groups ,and both lungs were completely resected under the condi‐tion of keeping mechanical ventilation .The vitro lungs were preserved by the way of the continuous infusion of a heart‐lung ma‐chine ,continuous pressure perfusion and single low temperature ,and collecting specimens according to the time point .HE staining was used to observe the morphological changes of vitro lung tissue .Immunohistochemistry and Western blot were used to detect the expression of AQP1 in vitro lung .Results HE staining found that as the time went by alveolar structure gradually collapsed ,in‐flammatory cells increased ,alveolar interval also gradually broadened and exudation could be seen in the alveolar cavity ;at the same time point ,organization structure of extracorporeal circulation group changed lighter than pressure perfusion group and low‐temper‐ature preservation group .In each experimental group ,the expression of AQP1 showed a trend of decline;at each time point ,the ex‐pression of AQP1 in extracorporeal circulation group was higher than pressure perfusion group ,and pressure infusion group was higher than that of low‐temperature preservation group .Conclusion The protective effect of the continuous infusion of a heart‐lung machine on vitro lung was better than continuous pressure perfusion and single low temperature .

Objective To investigate the effect and underlying mechanisms of ghrelin in a rat model of renal fibrosis. Methods Male Sprague-Dawley rats were divided into 4 groups , including sham operation +saline or brain gut peptide treatment group , model + saline or brain gut peptide treatment group. Unilateral ureteral obstruction (UUO) was established by left ureteral ligation. 7 days and 14 days after operation, the rats were sacrificed , while the kidney tissue of obstruction side was harvested for pathlogical changes through Masson coloration. Expression of α-smooth muscle actin (α-SMA), transforming growth factor beta1 (TGF-β1) and phosphorylated Smad3 (p-Smad3) in renal tissues were analyzed through immunohistochemistry. Expression of α-SMA and TGF-β1 mRNA was detected by real-time-PCR. Apoptosis kidneys cells were marked with TUNEL. Results Ghrelin inhibited renal fibrosis by reducing the production of collagen , restraining extracellular matrix (ECM) deposition and decreasing the expression of α-SMA. Meanwhile, ghrelin inhibited the accumulation of myofibroblasts by blocking the transforming growth factor-β1/Smad3 (TGF-β1/Smad3) signaling pathway. Moreover, ghrelin could attenuate renal tubular cell apoptosis induced by UUO injury. Conclusion Ghrelin can reduce renal fibrosis and renal cell apoptosis induced by UUO , demonstrating that ghrelin is a potent antifibrotic agent that may have therapeutic potential for patients with obstructive nephropathy.