Objective To compare the consistency and accuracy of determination of peripheral blood lymphocyte(PBL)interleukin-2(IL-2)with ELISA and evaluation of the expression of PBL IL-2 mRNA with real time PCR(RT-PCR).Methods In 20 kidney recipients,the PBL IL-2 level was determined with ELISA and the mRNA expression in PBL IL-2 was evaluated by real time fluorescence quantitative PCR(RT-FQ-PCR)before operation and C0/C2 time on the 7th day after operation.The same detection and evaluation were also done in 15 chronic allograft nephropathy(CAN)patients and 5 non CAN patients.Results IL-2 and IL-2 mRNA expression showed positive correlation before operation and 1 year later after operation,R2=0.58 and 0.4 respectively,both P0.05.On the 7th day after operation,IL-2 and the inhibition rate of IL-2 mRNA expression were(16.31?9.59)% and(69.92?7.10)%,respectively.A positive correlation existed between C2 IL-2 concentration and the inhibition rate of IL-2 mRNA expression,R2=0.56,P0.05.Also,IL-2 inhibition and PBL IL-2 mRNA inhibition showed no linear correlation.Conclusions The technique of determining PBL IL-2 by ELISA can be used for evaluating immunity status and immunosuppressive tolerance in patients with renal transplantation before the operation and at stable stage after operation.However,it should not be used for assessing perioperative immunity status and evaluating effect of cyclosporine.

Animals ; Cells, Cultured ; Female ; HSP70 Heat-Shock Proteins ; genetics ; metabolism ; Humans ; Lead ; pharmacokinetics ; toxicity ; Liver ; metabolism ; Male ; RNA, Messenger ; genetics ; Rats ; Rats, Wistar

Objective The purpose of this study was to analyze the alterations of T_1、T_2 lymphocyte subsets in the peripheral blood of patients with colorectal cancer. MethodsTwenty patients with primary colorectal cancer were enrolled into this study, T_1 and T_2 in the peripheral blood were evaluated by detecting the intracellular interferon-? and interleukin-4 production with 4-color flow cytometry. ResultsThe percentage of T_1 and T_2 in the peripheral blood of cancer patients was lower significantly than healthy controls [(36?11)% and 3.3(1.9)% vs. (46?12)% and 4.1(3.1)%](P

AIM:To investigate the principle of function of Stomach-Clearing Powder (Radix Rehmanniae, Radix Angelicae Sinensis, Cortex Moutan Rhizoma Coptidis, etc.) on clearing the stomach-heat. METHODS: 5% alcohol water were used as drink water of mice to make model. In the treatment group, "Stomach-Clearing Powder" was administed. Three weeks later, the anal temperature, the evacuation time of active carbon powder,stomach cAMP, SOD, MDA and the pathological changes in stomach and tongue were observed and recorded. RESULTS: "Stomach-Clearing Powder" could can obviously improve the pathological changes of the model group with stomach-heat syndrom. CONCLUSION: The established pathological model of stomach-heat syndrome accorded with traditional Chinese medicine; "Stomach-Clearing Powder" has the effect on clearing the stomach-heat on the experimental mouse.

Objective To investigate the effect of the genotypic analysis of donor from cardiac death donation on the initial dose of Tac for liver transplant recipients and provide individualized administration for the early use of Tac in liver transplantation patients.Method Thirty recipients with a different genotype of CYP3A5 from cardiac death donors were collected from March 2010 to February 2013.The matched recipients were randomly divided into experiment group and control group.There was an adjustment of initial doses of Tac according to the donors' different CYP3A5 genotypes in experiment group but not in control group.Result In experiment and control groups,the average Tac blood concentrations at the 7th day after operation were (7.47 ± 1.83) and (8.68 ± 5.14) ng/mL,and the percent of recipeints reaching the optimal Tac concentrations was 72.2％ and 38.9％,respectively (P＜0.05).In experiment and control groups,22.2％ and 55.6％ recipients needed adjustments of Tac concentrations respectively (P＜0.05).Conclusion Individualized adjustment of Tac initial doses of recipients according to cardiac death donors' different CYP3A5 genotypes was benefit for reaching optimal concentrations as soon as possible and could decrease the rate of rejection,and reduce the side effects of Tac.

Objective To construct the retroviral vector carrying novel gene mgt-16 and to investigate its expression in mouse mesenchymal stem cells C3H/10T1/2(10T1/2). Methods DNA sequences containing mouse novil gene mgt-16 was used as a template for PCR amplification of full length mgt-16 cDNA. Then the DNA fragment was cloned into pEGFP-N1 vector to produce pEGFP-N1-16 vector after T-A cloning with pMD18T plasmid and sequencing. The pEGFP-N1-16 vector was confirmed by PCR, restriction enzyme digestion and sequencing analysis. The retroviral vector, pLEGFP-N1-16, was constructed using retroviral vector, pLEGFP-N1, and pEGFP-N1-16 vector including mgt-16 gene. The pLEGFP-N1-16 vector was verified by restriction enzyme digestion, sequenced, and then transfected into packaging cell line Phoenix to prepare EGFP fused mgt-16 retrovirus particles, whichwere collected and used to infect 10T1/2 cells. G418 (400 (μg/mL) continuous selection was conducted to obtain 10T1/2 cell clones stably overexpressing EGFP fused mg-16. Fluorescence microscope was employed to determine the expression and subcellular localization of MGT-16 in Phoenix and 10T1/2 cells. Results A band of about 300 bp size was observed by agarose gel electrophoresis after PCR amplification for mgt-16 gene, and the result of sequencing showed that the sequence of insert fragment in T-A clones was identical to mg-16 gene reported in Genbank. PCR, restriction enzyme digestion and sequencing revealed that the pEGFP-Nl-16 plasmid was successfully constructed. Restriction enzyme digestion and sequencing revealed that the pLEGFP-Nl-16 plasmid was also successfully constructed. Phoenix and 10T1/2 cells overexpressing MGT-16 showed green fluorescence distribution in thecytoplasmic, especially around the perinucleararea. Conclusion We have successfully constructed a recombinant retroviral vector carrying novll gene, mg-16, and expressed it in mouse mesenchymal stem cells, which provides a basis for studying the role of novel gene mg-16 in mesenchymal stem cells.

In-vitro assay methods were established to evaluate transactivation and binding activity of compounds on peroxisome proliferator-activated receptor y (PPARγ). Firstly, plasmids were constructed for transactivation assay of PPARγ response element (PPRE) triggered reporter gene expression, and for cell-based binding activity assay of the chimeric receptor, which was fused with PPARγ ligand binding domain (LBD) and yeast transcriptional activator Gal4. Secondly, by using PPARy competitive binding assay based on time resolved-fluorescence resonance energy transfer (TR-FRET), affinities of compounds and drugs to PPARγ were evaluated. In application of these above methods, the PPARγ activating potency and characteristics of different compounds were evaluated, and a novel benzeneselfonamide derivative, ZLJ01, was found to have comparable binding activity and affinity with the well-known PPARy agonist, but lack of PPRE mediated transactivation activity. In preliminary study on in-vitro hypoglycemic activity, ZLJ1 was found to promote insulin-stimulated glucose uptake by liver cells. Therefore, we believe that combining transactivation and binding activity as well as affinity evaluation, the system could be used to screen non-agonist PPARγ ligand as anovel PPARγ modulator
Genes, Reporter ; Hepatocytes ; Hypoglycemic Agents ; chemistry ; Ligands ; PPAR gamma ; agonists ; chemistry ; Plasmids ; Response Elements ; Sulfonamides ; chemistry ; Transcriptional Activation

To investigate the effects of complex danshen solution and heparin on the changes of blood coagulation factors in rats with hemorrhagic shock, and to explore the therapy of coagulopathy by compound danshen solution, the rat model of hemorrhagic shock was set up, 40 SD rats were randomized into four groups: sham operation, shock, compound danshen solution and heparin groups, each group was composed of 10 SD rats. Plasma SFMC, TM, ATIII, D-D, t-PA, PAI levels and APTT were detected, incidences of bleeding complications between heparin and danshen group were compared. The results showed that plasma SFMC, D-D levels in shock group were higher but ATIII level in shock group was lower than that in sham operation group, compound danshen solution group and heparin group (P < 0.001), TM levels obviously increased in shock group and heparin group (P < 0.001). There was no significant difference between compound danshen solution and sham-operation groups. Plasma t-PA, D-D levels obviously increased after shock for 2 hours, PAI level reached the peak after shock for 4 hours, but t-PA decreased. After shock for 6 hours, plasma PAI descended, t-PA continually drop in, but PAI and D-D remained in higher levels. Plasma D-D level in heparin group was lower than that in shock group, t-PA level was higher than that in shock group, but there was no significant difference between in heparin and shock groups. Plasma t-PA, PAI and D-D levels in compound danshen solution group were lower than that in shock group. APTT of danshen group was lower than that of shock group and heparin group. Bleeding incidences was 30% in heparin group and 0% in danshen group, respectively. It is concluded that compound danshen solution may used to treat hypercoagulation and hyperfibrinolysis. In comparsion with heparin, danshen posses-ses advantages of safety with less bleeding complication and needs not tight monitor.
Animals ; Anticoagulants ; therapeutic use ; Blood Coagulation ; drug effects ; Blood Coagulation Factors ; metabolism ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Fibrin Fibrinogen Degradation Products ; metabolism ; Heparin ; therapeutic use ; Male ; Partial Thromboplastin Time ; Phytotherapy ; Plasminogen Activator Inhibitor 1 ; blood ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Salvia miltiorrhiza ; chemistry ; Shock, Hemorrhagic ; blood ; drug therapy ; Thromboplastin ; metabolism ; Tissue Plasminogen Activator ; blood

Anti-Bacterial Agents ; therapeutic use ; Antigens, CD20 ; metabolism ; CD3 Complex ; metabolism ; CD4 Antigens ; metabolism ; Fever ; drug therapy ; virology ; Herpesvirus 6, Human ; isolation & purification ; Humans ; Lymph Nodes ; pathology ; virology ; Lymphadenitis ; drug therapy ; metabolism ; pathology ; virology ; Male ; Middle Aged ; Roseolovirus Infections ; drug therapy ; metabolism ; virology

Objective To investigate the development of absorption atelectasis in healthy adult volunteers breathing 100% oxygen.Methods Six healthy volunteers aged 31-36 yrs weighing 60-80 kg were recruited into this study.Chest computed tomograph(CT)of the layer of interventricular septum was performed at the end of normal expiration(FRC)at 6 time points:(1)the baseline(T_1);(2)after 5 min maximal expiration(close to residual volume)(T_2);(3)immediately after 10 maximal inspiration and expiration(T_3);(4)after breathing 100% O_2 through face mask at tidal volume for 30 min(T_4);(5)after breathing 100% O_2 at maximal expiration for 5 min(T_5)and(6)breathing room air deeply 10 times(T_6).The area of atelectasis and poorly ventilated lung were expressed as percentage of the total lungs.Results There was no atelectasis or poorly ventilated lung at T_1. At T_2 the poorly ventilated lung accounted for 22.9%?5.0%,but there was no atelectasis.There was no atelectasis and poorly ventilated lung at T_3 and T_4.At T_5 atelectasis accounted for 4.5%?1.1% which was reduced to 0.9%?0.4% at T_6.Conclusion Breathing 100% oxygen at reduced lung volume can result in atelectasis.