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.

OBJECTIVETo investigate the protect effects of sodium ferulate (SF) on the daunormbicin(DNR-induced cardiotoxicity in juvenile rats.

METHODSForty male juvenile SD rats were randomly divided into control group (Control), daunorubicin group (DNR), sodium ferudate treatment group (DNR + SF), sodium ferudate group (SF) (n = 10) . Juvenile rats were intraperitoneally treated with DNR (2.5 mg/kg every week for a cumulative dose of 10 mg/kg) preparation immature myocardial injury model in presence with SF (60 mg/kg) oral treat- ment for 25 days. The left ventricular pressure and its response to isoproterenol were measured using left ventricular catheter. Rat myocardium myocardial pathology specimens and ultrastructure changes were also observed. The expression of cardiac Troponin I (cTNI) was detected by Western blot and RT-PCR. Results: SF treatment could inhibit the decreasing of heart rates induced by DNR damage (P < 0.05); it could increase the left ventrivular end diastolic pressure(LVEDP), heart rate, the maximal left ventrivular systolic speed(LVP + dp/dtmax) and the maximal left ventrivular diastolic speed (LVP-dp/dtmax) responding to isoproterenol stimulation(P < 0.01); SF also could improve the myocardial ultrastructure injuries and inhibit the decreasing of cTNI expression caused by DNR damages (P < 0.05).

CONCLUSIONSF treatment could alleviate the decreasing of cardiac reservation induced by DNR damages in juvenile rats, which might be related to its reversing the effects on the cardiac systolic and diastolic function injuries and its inhibiting effects on the decreasing of cTNI expression caused by DNR. The mechanism of SF preventing daunorubicin-induced cardiotoxicity in juvenile rats is relevant to inhabited cardiac Troponin I expression.

Animals ; Blood Pressure ; Cardiotoxicity ; drug therapy ; Coumaric Acids ; pharmacology ; Daunorubicin ; toxicity ; Heart ; physiopathology ; Heart Rate ; Isoproterenol ; Male ; Myocardium ; pathology ; Protective Agents ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Troponin I ; metabolism

Animals ; Animals, Newborn ; Chronic Disease ; Daunorubicin ; adverse effects ; Disease Models, Animal ; Heart Failure ; chemically induced ; Rats ; Rats, Sprague-Dawley

BACKGROUND: There is no effective method to identify liver cancer stem cells until now, which has become one of the major challenges in this field. OBJECTIVE: To explore a more effective and suitable way for the identification of liver cancer stem cells in hepatocellular carcinoma cell lines. METHODS: Firstly, the single cell separation technique was used to obtain single cells, which were seeded into 96-wel plates one by one. Secondly, cell sublines derived from single cell colonies (tumorigenic colonies) were selected and obtained. At last, the cells from these clones were transplanted into the forelimb armpits of nude mice to observe the tumorigenic ability. RESULTS AND CONCLUSION: The number of holes of single cells obtained was 371 by the single cell separation technique, and the success rate was 96.4%. According to the growth of cell clone, tumorigenic colonies were selected to be transplanted to the forelimb armpits of nude mice, and the tumor formation rate of the colonies was 100%. The identification model of single-cell-transplant-tumorigenic-test for liver cancer stem cells is confirmed to be preliminarily established, which lays the technology and methodological basis for the follow-up research.

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 explore the clinical characteristics and outcome of group B streptococcus (GBS) induced neonatal meningitis and to provide the guide for early diagnosis and appropriate treatment.Methods A retrospective chart review was performed.A total of 19 cases of neonatal purulent meningitis caused by GBS and 22 cases of neonatal purulent meningitis caused by Escherichia coli were identified in the NICU of Guangzhou Women and Children's Medical Center from Nov 1,2011 to Apr 31,2014.The clinical features,treatments and clinical turnover were analysed.Results GBS meningitis accounted for 24.7％ (19/77) of total bacterial positive cultures of blood or cerebral spinal fluid.The average time of progression to early-onset GBS meningitis of 6 early-onset cases mainly complaining of anhelation and groan,was (11.80 ± 11.34)h,and 83.3％ present within 24 hours;the main initial clinical symptoms of 13 late-onset cases[mean age (17.85 ± 7.77) d] were fever.Peripheral blood C-reactive protein concentration of GBS meningitis was significantly higher than that of Escherichia coli meningitis [(154.43 ± 88.64) mg/L vs.(67.52 ± 64.23) mg/L,P =0.001].Compared with Escherichia coli meningitis,the average length of stay in hospital and the recovery time of abnormal cerebral spinal fluid in neonates with GBS infection were both extended by more than 10 days.Conclusion The clinical manifestations of neonatal purulent meningitis caused by GBS are usually non-specific.It is associated with longer hospitalization and recovery time of abnormal cerebral spinal fluid.Antepartum prophylaxis,early diagnosis and therapy are vital for reducing the incidence of complications and mortality of neonatal GBS purulent meningitis.

Objective To evaluate the protective effect and mechanism of luomaishutong(LMST)on acute myocardial ischemia-reperfusion injury in rabbits.Method Twenty-four New Zealand white rabbits were randomly divided into 3 groups:the LMST group,the control group and the sham-operated group.The AMI reperfusion model was established by removing the blockade after the occlusion of coronary artery for 2 hours.The changes of hemodynamics,oxygen free radical and clearance system were measured in all rabbits.Results (1)Compared with the control group,left ventricular systolic pressure(LVSP)and maximal changing rate of left ventricular innner pressure(?dp/dt_(max))increased remarkably in the LMST group after reperfusion,meanwhile, LVEDP decreased significantly.(2)In the control group,MDA level of cardiocyte was noticeably higher,while SOD and NOS levels were lower than in the sham-operated group.Compared with the control group,MDA level in the LMST group was significantly lower.Furthermore,SOD and NOS levels were higher in LMST group,and the infarcted area was smaller in the LMST group as well.Conclusions LMSF can protect myocardium after ischemia-repcrfusioninjury and improve cardiac function through inhibiting induced by oxygen free radicals.

BACKGROUND:Until now, little has been reported on establishment of pancreatic stem cell strains and lines,and the purification of pancreatic stem cells is difficult since the cell line establish rate is low.OBJECTIVE:To explore a more rational and effective technique of in vitro separation and continuous passage of pancreatic stem cells, with the hope to establish cell strains and even cell lines and to lay the foundation for the follow-up study of pancreatic stem cells in the treatment of diabetes.METHODS:Firstly, Percoll discontinuous density gradient centrifugation method was applied to separate the mouse pancreatic endocrine portion from the exocrine portion, then to obtain cell strains with highly proliferative ability and low differentiation from pancreatic endocrine portion-the islet. We used mouse embryonic fibroblasts treated with mitomycin C as a feeder layer, for in vitro continuous culture of islet-derived pancreatic stem cells under feeder layer conditions until they were transferred to the 30th passage to establish cell lines. Then pancreatic stem cell line derived from pancreatic islet was detected and identified by a series of tests including growth characteristic test, morphological observation, related molecular marker identification and differentiation characteristic identification.RESULTS AND CONCLUSION:In the continuous process of passage, pancreatic stem cells showed active proliferative ability, and maintained the typical morphological characteristics of stem cells and expression of pancreatic stem cell marker-Nestin. After induction, pancreatic stem cells showed insulin gene expression,reflecting their differentiation potential. Therefore, under the condition of feeder layer, the pancreatic stem cell line derived from Kunming mice was successfully established and the related identification was completed,which lays the foundation for the following research.

Exosomes are bilayer-lipid membrane nanovesicle from almost all living cell types which are in-volved in intercellular substance transporting and signaling communication .Exosomes are 30 ~120 nm in diameter , can transfer bioactive molecules including DNA , RNA, microRNA, protein as well as lipids derived from parents ' cells to re-cipient cells by body fluids , and specifically influence their physiological or pathological conditions .Leukemia is due to malignant proliferation of hematopoietic stem and progenitor cells .It was reported that leukemia cells derived exosomes play a key role in disease progression , drug resistance , and predict prognosis .This paper will outline the role of exosomes de-rived from leukemia cells and provide important information to help explore the molecular pathogenesis , biomarker as well as therapeutic target of leukemia .

OBJECTIVETo explore the effects of progesterone (PROG) on learning and memory and P2X7 receptor expression in the hippocampus after global cerebral ischemia/reperfusion (I/R) injury in rats.

METHODSForty-eight male SD rats were randomly divided into four groups (n = 12) that were normal group, sham-operated group, I/R group and I/R+ PROG group. The global cerebral I/R injury models were established by improved Pulsinelli's four vessel occlusion, the learning and memory were evaluated by Y-maze, the expression of P2X7 receptor protein in the hippocampus were detected by the immunofluorescence, the activity of superoxide dismutase (SOD) in the hippocampus were detected with hydroxylamine oxidation method, the content of malondialdehyde (MDA) in the hippocampus were detected with pen-thiobarbituric acid method.

RESULTSThere was no significant difference of the learning and memory and positive expression cells of P2X7 receptor protein and the activity of SOD and the content of MDA in the hippocampus between normal group and sham-operated group. Compared with sham-operated group, the learning and memory of I/R group were significantly decreased (P < 0.01), Compared with I/R group, the learning and memory of I/R + PROG group were significantly increased (P < 0.05). Compared with sham-operated group, the positive expression cells of P2X7 receptor protein in the hippocampus of I/R group was significantly increased (P < 0.01), Compared with I/R group, the positive expression cells of P2X7 receptor protein in the hippocampus of I/R + PROG group was significantly decreased (P < 0.01). Compared with sham-operated group, the activity of SOD in the hippocampus of of I/R group was significantly decreased (P < 0.01), and the content of MDA in the hippocampus of I/R group was significantly increased (P < 0.01). Compared with I/R group, the activity of SOD in the hippocampus of I/R+ PROG group was significantly increased (P < 0.05), and the content of MDA in the hippocampus of I/R + PROG group was significantly decreased (P < 0.01).

CONCLUSIONPROG could improve the learning and memory ability following global cerebral ischemia/reperfusion injury in rats, the protective mechanism might be associated with down-regulating the expression of P2X7 receptor protein and attenuating oxygen-derived free radicals in the hippocampus.

Animals ; Brain Ischemia ; metabolism ; psychology ; Hippocampus ; drug effects ; metabolism ; Male ; Malondialdehyde ; metabolism ; Maze Learning ; drug effects ; Progesterone ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Receptors, Purinergic P2X7 ; metabolism ; Reperfusion Injury ; metabolism ; psychology ; Superoxide Dismutase ; metabolism