BACKGROUND: To avoid acute rejection,it is necessary to use imunosuppressive drug regimen for long term to control immune state.However,imunosuppressive drug regimen of allogenic organ transplantation increases infection incidence of recipients,and induction of allograft immunological tolerance might be an ideal method for solving these problems.The long-term immunologic tolerance has been able to be induced in the experimental rodent models.Among these protocols,donor bone marrow cell (DBMC) infusion exerts an important role in the induction of allograft immunological tolerance.OBJECTIVE: To investigate effects of combination of cyclosporine A (CsA) with DBMC infusion on heterotopic rat cardiac allograft survival time.DESIDN: A randomized controlled animal experiment.SETTING: Renal Disease Center,First Affiliated Hospital of Zhejing University School of Medicine.MATERIALS: This study was performed at the Laboratory Animal Center,Zhejiang University School of Medicine between March 2002 and December 2005.Inbred male Lewis rats (n=40,serving as donors) and male BN rats (n=60,serving as recipients) of SPF grade were used in this study.The protocol was approved by the Hospital's Ethic's Committee.METHODS: Forty rats prepared for heterotopic rat cardiac allograft were randomly divided into 4 groups,with 10 rats in each: control group,in which,rats received no treatment,CsA group,in which,rats received CsA infusion for 7 days successively; CsA +DBMC group,in which,rats received DBMCs during and 6 days after the surgery and additional 7 successive days of CsA infusion,and a DBMC group,in which,rats received DBMCs infusion during and 6 days after the surgery.In addition,BN rats that received beterotopic rat cardiac allograft served BN controls.The survival time of heteroropic rat cardiac allograft was investigated.Serum interleukin-2 level and tumor necrosis factor-α mRNA expression level in the transplanted cardiac allograft were measured. The percentage of antigen presenting cells (APC) from donor,CD3+CD25+ cells,CD4+CD25+ cells,CD86+ cells,and the ratio for CD4+CD45RC+ and CD4+CD45RC- in the recipient peripheral blood karyocytes were measured by flow cytometry 6,12 and 18 days after surgery.MAIN OUTCOME MEASURES: The survival time of beteruropic rat cardiac allograft,serum interleukin-2 (IL-2)level,tumor necrosis factor- α (TNF- α ) rnRNA expression level, rejection grading,the percentage of DBMCs in the recipient peripheral blood karyocytes,CD3+CD25+ cells,and CD4+CD25+ cells,as well as CD86 expression,and the ratio for CD4+CD45RC+ and CD4+CD45RC.RESULTS: Forty Lewis male rats and sixty male BN rats were all included in the final analysis. The heterotopic rat cardiac allograft survival time was longer in the CsA +DBMC group than in the control group and DBMC group (P ＜ 0.05). Serum IL-2 level and TNF- α mRNA expression were respectively lower in the CsA +DBMC group than in the control group and DBMC group ( P ＜ 0.05).The rejection was milder in the CsA +DBMC group than in the remaining 3 transplantation groups.In the CsA +DBMC group,CD 86 expression in the recipient peripheral blood karyocytes was markedly inhibited,and 6 and 12 days after surgery,the ratio for CD4+CD45RC+ and CD4+CD45RC- and the percentage of CD3+CD25+ were respectively lower compared to control group and DBMC group.DBMCs in the recipient peripheral blood karyocytes were more in rats that received DBMC infusion compared to rats that received no BDMC infusion.CONCLUSION: Short-term CsA treatment combined with DBMC infusion can lower acute rejection of heterotopic rat cardiac allograft and prolongssurvival time of cardiac allograft.

Objective To investigate the changes of lead,zinc,copper,iron and calcium in blood of chronic poisoned infantal mice.(Methods) Forty-eight 21 day-old kunzea mice were randomly divided into 4 groups,each having 12 mice.Distilled water group was as control group and other three lead acetate poisoning groups had a dose of 10,20,40 mg/kg,respectively.The poisoning was carried out by lavage once a day,and consecutively for 46 days.Eyeballs of mice were picked then for blood sampling,and BS trace element analysis grapher was used to determine level of lead,zinc,copper and iron.Level of calcium was measured by Dimentional-RXL auto-biochemistry analysis meter.Results The lead and zinc levels in poisoned mice blood were increased with increasing lead acetate level administration,while zinc level changed inversely with lead acetate level.Significant differences were shown among control group and poisoning groups in terms of lead(P0.05).Conclusion Lead posioning can lead to zinc decreasing and copper(increa)-sing,which suggests that zinc works as a poential antidote of lead poisoning.

Objective To observe the value of ultrasonography, MR and MSCT in diagnosing intravitreous autologous eyelashes. Methods Thirty-two New Zealand rabbits were averagely divided into 4 groups (A, B, C and D) randomly, and 1, 5, 10 eyelashes were implanted into vitreous body of rabbits respectively in group A, B, and C, whereas rabbits in group D (the control group) were only exposed to sham operation without implanting any eyelashes. After one week, all rabbits underwent ultrasound, MR and MSCT examination, then the specimens of vitreous bodies were obtained and observed pathologically. Results All the implanted intravitreous eyelashes were displayed with ultrasound. Intravitreous eyelashes in group A were not detected with MRI, whereas in 3 rabbits of group B and all of group C were displayed on T2WI, T2*WI and SWI sequences. MSCT detected intravitreous eyelashes in only 6 experimental animals. Conclusion Ultrasonography should be considered as the first choice for diagnosis of intravitreous autologous eyelashes. Different sequences of MRI have various advantages in diagnosing intravitreous autologous eyelashes, whereas the diagnostic value of MSCT is limited.

Objective:To investigate the relationships between the expression levels of chemokines CCL19 and CCL21 in serum of patients with RA and interstitial lung disease,as well as the correlation between the chemokines and the clinical parameters.Methods:Patients hospitalized in the First Hospital of China Medical University from May 2016 to March 2017 were enrolled.Serum levels of CCL21 and CCL19 in the 62 patients with RA,among them,34 cases without pulmonary interstitial lesions while 28 cases with lung interstitial diseases,18 cases ⅡP and 20 health control were detected with enzyme-linked immunosorbent assay (ELISA).One-way ANOVA,LSD-t test,Kruskal-Wallis test,Pearson and Spearman correlation analysis were used for statistical analysis.Results:The levels of CCL21 and CCL19 were both higher in the RA patients than the health controls(P＜0.05;P＜0.01),The concentrations of CCL21 in serum of the RA patients with ILD were higher than thoes without ILD,higher than the health controls (P＜0.05;P＜0.01),serum levels of CCL21 in the ⅡP patients were higher than the health controls (P＜0.01),too.The serum levels of CCL21 in men were higher than women(P＜0.05).Serum CCL21 levels were negatively correlated with albumin and BMI(r=-0.280,P＜0.05;r =-0.605,P＜0.05).Levels of CCL19 were negatively correlated with the levels of C4 (r =-0.326,P＜0.05),and positively correlated with the levels of D-Dimer(r =0.592,P＜0.05).Conclusion:Chemokines CCL21 and CCL19 are highly expressed in serum of patients with RA,they may be involved in the pathogenesis of RA.Furthermore,CCL21 may has a correlation with the lung interstitial lesion in RA.

Objective To study the distribution of pathogens,the positive time and drug resistance of pathogenic bacteria by blood culture of adult patients,in order to provide the basis for the early clinical discovery and treatment of bacteremia. Methods 3 537 specimens of adult blood culture were collected from July 2016 to December 2016,then identified the posi-tive bacteria strains,and analysed the antimicrobial susceptibility.Results In 3 537 specimens of adult blood culture,485 positive samples were detected,and the positive rate was 13.7%(485/3 537).Including 203 cases(41.9%)of both aerobic and anaerobic positive bottles,220 cases(45.3%)of aerobic positive bottles,and 62 cases(12.8%)of anaerobic positive bottles.About pathogens,229 specimens were gram-negative bacteria strains,accounting for 47.2%.The great majority of bacteria was E.Coli,Klebsiella pneumoniae,Pseudomonas aeruginosa,and they all showed sensitivity to imipenem.202 specimens were gram-positive bacteria strains,accounting for 41.7%,mainly on Staphylococcus aureus and Staphylococcus epidermidis,they all showed sensitive to vancomycin.54 strains were Fungi,accounting for 11.1%.For analysis of 203 ca-ses of aerobic and anaerobic both positive bottles,the results showed that:there were 121 cases of gram-negative bacteria strains,95(78.5%)specimens anaerobic jar to positive time earlier than aerobic bottle to positive time,26(21.5%)speci-mens aerobic bottle jar to positive time earlier than anaerobic bottles to positive time.78 cases of gram positive bacteria,an-aerobic jar to earlier than aerobic bottle to positive time had 45 strains,accounting for 57.7%.Aerobic bottle to positive time earlier than anaerobic bottles to positive time had 33 strains,accounting for 42.3%.Fungi,a total of 4 strains,50% each. Positive pathogens were mainly distributed in I,emergency surgery,respiratory medicine department.Conclusion Pathogen-ic bacteria isolated from the adult blood culture was given priority to gram-negative bacteria,pathogenic bacteria species and drug susceptibility difference was obviously.Clinicians should be combined with blood culture and drug susceptibility results of use of antimicrobial drugs to patients.

Alisma orientalis is a traditional herb medicine commonly used in clinical. With the increasing report of its toxicity in clinical, the renal toxicity of Alisma orientalis has got gradually attention. This paper systematically reviews the research on the chemical material basis of Alisma orientalis including its chemical composition and toxicity of ingredients; and also declares its toxic ingredients and targets according to Network toxicology. Based on the controversy on renal toxicity of Alisma orientalis, we analyzed the possible reasons that may be associated with renal toxicity. It might be associated with the differences of the material basis composition and regulatory toxicology network, differences in employed processing technology, the metabolic function leading to accumulation of compounds, dosage and duration of the experiment and compatibility. The review provides possible reference and ideas for the quality control and rational use of Alisma orientalis.
Alisma ; chemistry ; toxicity ; Animals ; Drugs, Chinese Herbal ; chemistry ; toxicity ; Humans ; Molecular Structure

This study was purposed to explore the expression of p57kip2 in the bone marrow of patients with de novo myelodysplastic syndrome (MDS) and its role in MDS pathogenesis, as well as the relationship between the expression of p57kip2 and SDF-1/CXCR4 signal. The expression of p57kip2 and CXCR4 in 67 de novo MDS patients was measured by real-time quantitative PCR. The percentage of CD34(+) cells in the bone marrow from MDS patients was measured by flow cytometry. 18 healthy volunteers were recruited for control. The effect of SDF-1 on p57kip2 expression in bone marrow mononuclear cell (BMMNC) from MDS or normal controls was investigated in vitro, and difference between them was compared. The results showed that low-risk MDS and high-risk MDS displayed a significant reduction of p57kip2 mRNA expression in BMMNC compared with that in control group (P < 0.001) and there was a negative correlation between p57kip2 expression and percentage of CD34(+) (r = -0.458, P < 0.001); the patients with abnormal karyotype showed lower expression of p57kip2 gene, compared to patients with normal karyotype (P = 0.045). Although the expression of CXCR4 had no difference between MDS patients and normal controls, a positive correlation between p57kip2 and CXCR4 in MDS patients was still found (r = 0.609, P < 0.001). Moreover, SDF-1 increased p57kip2 expression in normal BMMNC in dose-dependent manner, but BMMNC from MDS patients showed no response to SDF-1. SDF-1-induced p57 expression was blocked by AMD3100. It is concluded that the low expression of p57 gene in MDS may play a role in the pathogenesis of MDS. Furthermore, SDF-1-induced p57kip2 expression in BMMNC, and the decreasing response of BMMNC to SDF-1 may contribute to the low expression of p57kip2 in MDS patients.
Case-Control Studies ; Chemokine CXCL12 ; metabolism ; Cyclin-Dependent Kinase Inhibitor p57 ; genetics ; metabolism ; Flow Cytometry ; Humans ; Myelodysplastic Syndromes ; genetics ; metabolism ; Receptors, CXCR4 ; metabolism

OBJECTIVETo study the methylation status of p73 gene promoter in patients with myelodysplastic syndrome (MDS) and explore its significance with clinical prognosis.

METHODSMethylation of p73 promoter was detected in bone marrow cells from 135 MDS patients and 13 healthy controls by methylation-specific PCR (MSP). The results of MSP were confirmed by bisulfite sequencing. The expression of p73 mRNA was detected by real-time quantitative PCR. Primary bone marrow cells from MDS patients were treated with decitabine, the changes of p73 methylation status and p73 mRNA expression were measured. The role of p73 methylation in the prognosis of MDS and the correlated clinical data were explored.

RESULTSp73 hypermethylation was present in 37.04% of MDS cases and patients with high risk MDS (RAEB-1 and RAEB-2) exhibited a significantly higher frequency of p73 methylation than that of low risk MDS (58.8% vs 29.7%, P = 0.002). The expression of p73 mRNA in the methylated group was decreased compared to that of the unmethylated group (P = 0.032). Decitabine treatment decreased the level of p73 methylation and increased the level of p73 transcripts. Patients with p73 methylation progressed rapidly to AML (P < 0.001) and had shorter survival (P = 0.002) than those who did not have p73 methylation. In the multivariate Cox regression model, BM blast and p73 methylation status emerged as independent prognostic factor for overall survival and leukemia free survival.

CONCLUSIONp73 gene methylation is common in patients with MDS and may indicate poor prognosis. p73 may be a therapeutic target in MDS.

Aged ; Case-Control Studies ; DNA Methylation ; DNA-Binding Proteins ; genetics ; Female ; Humans ; Male ; Middle Aged ; Myelodysplastic Syndromes ; genetics ; Nuclear Proteins ; genetics ; Prognosis ; Promoter Regions, Genetic ; Tumor Protein p73 ; Tumor Suppressor Proteins ; genetics

Development of the disease is the result of several factors involved in biological network changes. The nature of drug intervention is to regulate these pathological changes to the normal range. Advantages of traditional Chinese medicine (TCM) are to integrally and systematically regulate this biological networks and systematic pathology through multi-targets, multi-levels, multi-channels. Structural components TCM provides the controlled and precise basis "substance" for this regulation and also to clarify the "truth" of the nature of the regulation by the network pharmacology. Network pharmacology provides new strategy for the research on mechanism of structural components TCM. This study not only reflects the overall characteristics of the development of the disease, but also fully embodies the essence of TCM for preventing and treating diseases through changing traditional model on "one drug, one gene, one disease". This paper explores systematically the integration essence, features and research strategies of structural components TCM and the network pharmacology, understand the interaction of structural components TCM and body from the perspective of the overall concept of improving or restoring the balance of.biological networks. It is effective measure to reveal the structure of a multi-component for regulating biological networks mechanisms, and also provide new ideas and methods for further scientific research and innovation of structural component TCM.
Drug Interactions ; Drug Therapy ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Gene Regulatory Networks ; drug effects ; Humans ; Medicine, Chinese Traditional

To study the pharmacokinetics, excretion characteristics and plasma protein binding rate of asperosaponin VI (A-VI) and its active metabolite hederagenin (M1). A-VI and M1 concentrations in plasma, bile, urine and feces were determined by established LC-MS/MS to calculate the pharmacokinetic parameters. The plasma protein binding rate of A-VI was determined by equilibrium dialysis method. the double peaks was observed in the A-VI plasma concentration-time curve, after rats were orally administered with low, medium and high doses of A-VI (0.03, 0.09, 0.27 g x kg(-1)). The Cmax1 and Cmax2 of A-VI were (28.88 +/- 49.78) and (4.480 +/- 1.872) microg x L(-1), (35.19 +/- 23.53) and (22.11 +/- 16.15) microg x L(-1), (73.37 +/- 37.28) and (132.2 +/- 160.7) microg x L(-1), respectively. The AUC0-t, of A-VI were (43.21 +/- 37.32), (133.9 +/- 102.5) and (779.6 +/- 876.9) microg x h x L(-1), respectively. The t1/2 of A-VI were (3.3 +/- 0.8), (3.2 +/- 2.3) and (4.5 +/- 1.2) h, respectively. The Cmax of M1 were (16.03 +/- 9.336), (26.41 +/- 11.95) and (28.71 +/- 5.874) microg x L(-1), respectively. The AUC0-t, of M1 were (105.6 +/- 73.60), (260.0 +/-153.9) and (323.1 +/- 107.9) microg x h x L(-1), respectively. The t1/2 of M1 were (4.1 +/- 3.4), (4.4 +/- 2.3), (3.9 +/- 0.9) h, respectively. No significant gender difference was found in the in vivo pharmacokinetics of A-VI and M1. There was no accumulation of A-VI and M1 after multiple administrations of A-VI (0.09 g x kg(-1)). After oral administration of A-VI, the double peaks were also observed in biliary and urinary excretion rate-time curves for A-VI. M1 was detected in the feces samples at 6 h after oral administration. The average plasma protein binding rate of A-VI was 92. 9% in rats.
Administration, Oral ; Animals ; Area Under Curve ; Bile ; metabolism ; Drugs, Chinese Herbal ; metabolism ; pharmacokinetics ; Female ; Male ; Plants, Medicinal ; Protein Binding ; drug effects ; Rats ; Saponins ; blood ; metabolism ; pharmacokinetics ; urine