Objective To develop a new rabbit model of myocardial infarction and assess left ventricular function and the survival rates.Methods A total of 30 adult rabbits were equally and randomly assigned into improved（n =15）and traditional model groups（n =15）.The rabbits were not intubated and ventilated in improved model group.A thoracotomy was performed along the left sternum by small nick with rib and pleura cavity integrity.The left anterodecendant arteries（LAD）of adult rabbits were ligated to establish animal models of my-ocardial infarction by a 60-min occlusion of the LAD,followed by reperfusion.In another group,rabbits were intubated and ventilated with rib abscised during thoracotomy inversely.Echocardiographic assessment was performed 3 weeks after myocardial infarction and survival rates were analyzed.Results Echocardiography showed that there was a sizable reduction in systolic and diastolic function in both groups with cardiac function significantly decreased 3 weeks later.Compared with traditional model group,improved method significantly increased the survival rate of rabbit model with myocardial ischemia.Conclusion This improved method is applicable to make an animal model of my-ocardial ischemia in rabbit.We established a simple,rapid,and effective method to mimick myocardial infarction in rabbit.

BACKGROUND:Bone marrow mesenchymal stem cells(BMSCs) have the potential to differentiate into various cells and growth factor plays a key role in cell engraftment, survival, and differentiation.OBJECTIVE:To observe the effects of hepatocyte growth factor(HGF) and insulin-like growth factor(IGF) combined with stem cell transplantation on cardiac function in rabbits after myocardial infarction.DESIGN, TIME AND SETTING:The randomized controlled animal study was performed at the Animal Experimental Center, China Medical University from November 2006 to March 2008.MATERIALS:A total of 28 healthy adult rabbits, of both genders, weighing 2.0-2.5 kg were used in this study.METHODS:Rabbit BMSCs were cultured in vitro.The left anterodecendant arteries(LAD) of adult rabbits were ligated to establish animal models of myocardial infarction by a 60-min occlusion of the LAD, followed by reperfusion.A total of 28 adult rabbits were equally and randomly assigned into 4 groups.In the combination group, rabbit models were injected with 8 ?L DMEM, supplemented with 5?1011/L BMSCs 40 ?L, 150 ?g/L HGF and 200 ?g/L IGF-1 at 3 days following model induction.In the stem cell group, rabbit models were infused with 5?1011/L BMSCs 40 ?L.In the growth factor group, rabbit models were injected with 8 ?L DMEM containing 150 ?g/L HGF and 200 ?g/L IGF-1.In the control group, rabbit models were treated with the same volume of DMEM.Three dimensional(3D) full volume image of the left ventricle was measured by IE33 model ultraphonic apparatus with 3D detecting head.MAIN OUTCOME MEASURES:The following parameters were measured:results of BMSC fluorescent labeling, end diastolic dimension, end systolic volume and ejection fraction of the left ventricle in each group at 6 weeks following model induction.RESULTS:A total of 28 adult rabbits were included in the final analysis.BMSCs adhered, with various morphology.Following 1 week, adhered cells gradually formed cell colony, with the presence of spindle-shape.At 2 weeks, about 80%-90% cells were confluent.Under a fluorescence microscope, DAPI-positive cells were found in the combination and stem cell groups.Positive cells were scattered in the rabbit myocardium, and surrounding the scar, along cardiac muscle fiber direction.Compared with the control and growth factor groups, cardiac function was significantly improved in the combination and stem cell groups(P

Background BackgroundMeningothelial cells (MECs) are major cell type in the meningeal sheath around optic nerve,which form a fluid barrier between optic nerve and the cerebral spinal fluid.The impairment of the cerebral fluid-optic nerve barrier probably affects the balance of cerebral fluid components.Currently,the investigation on the role of MECs in neuropathy is less performed.Objective This study attempted to explore hypoxia-induced function changes of MECs,and to shed a new clus for the future research of optic nerve disorders.Methods Human MECs strains were cultured in vitro and cell suspension was prepared with the cell densities of 2.5 ×103/hole,5.0× 103/hole and 1 x 104 /hole,respectively.The suspensions of 100 μl were separately collected to incubate in 96-well plates and cultivated for 2 days in 21％ O2(normoxia group) or 1％ O2(hypoxia group).MTS was used to detect and compare the proliferative value (A490) of MECs between the normoxia group and the hypoxia group.The changes of MECs diameter and volume were measured by CASY1 assay.ATP product in the cells after MECs exposed to different oxygen environments with or without substrate (100 mmol/L pyruvate and 100 mmol/L malate) for 1,2 days were assayed by Luminometer method.The expression and distribution of cytochrome C in the cells of the normoxia group and the hypoxia group were determined by immunofluorescence.Results A490 of MECs in the 2.5× 103/hole,5.0× 103/hole and 1 × 104/hole were 0.399±0.009,0.393±0.009 and 0.496±0.026 in the hypoxia group,which were lower than 0.424±0.131,0.413±0.111 and 0.537±0.021 in the normoxia group (t =3.777,P =0.004 ; t =3.251,P =0.009 ; t =3.037,P =0.013).Compared with the normoxia group,the diameter and volume were significantly increased in the hypoxia group ([20.970 ±0.127] μm vs.[21.198 ±0.048] μm,t =-3.762,P=0.006; [5805±73] fl vs.[6026±106] fl,t=-4.124,P=0.002).ATP products were (0.900±0.225)mmol/(L· g) and (0.952± 0.075) mmol/(L · g) in the hypoxia group and the hypoxia+substrate group,which were significantly lower than (1.389±0.145) mmol/(L · g) and (1.401±0.122) mmol/(L · g) in the normoxia group and the normoxia +substrate group (P =0.001,0.002,0.001).Immunofluorescense staining showed that the green fluorescence of cytochrome C located at mitochondria of MECs in the normoxia group,but in the hypoxia group,cytochrome C distributed in the cytoplasm extensively.Conclusions Hypoxia induces malfunction of MECs,which might impact the intact of the cerebral spinal fluid-optic nerve barrier and therefore influence the microenvironment of the subarachnoid space and neuronal function.

Objective To explore the efficacy of multiple target therapy in treatment of patients with chronic moderate and severe IgA nephropathy with hyperuricemia.Methods Seventy-six patients with chronic progressive moderate and severe IgA nephropathy with hyperuricemia were enrolled the current study and randomly divided into observation group and control group.Patients in control group were treated with allopurinol,prednisone,benner pury and valsartan,while those in observation group were treated with urokinase,mycophenolate mofetil besides the basis of control group for 6 months.The blood uric acid (UA),24 h urine protein,mean arterial pressure (MAP) and creatinine clearance rate (Ccr) were determined and analyzed.Results The levels of UA,24 h urinary protein,MAP and Ccr in observation group and control group were same before treatment (P ＞ 0.05).After 6 months treatment,the levels of UA,24 h urine protein,MAP and Ccr in observation group were (413.7 ± 90.7) μmol/L,(1.15 ± 0.57) g/L,(87.7 ± 10.6) mmHg and (81.9 ± 3.7) ml/min respectively,significantly different from those of the control group ((369.6 ± 67.2) μ mol/L,(0.77 ±0.51) g/L,(81.6 ±12.3) mmHg and (86.4 ±6.8) ml/min;t =2.219,2.802,2.132,3.230;P ＜0.05).The rate of adverse reactions in two groups was not significantly differnent(9.7％ (3/31) vs 9.1％ (3/33) ; x2 =0.006,P =0.936).Conclusion Multiply target therapy is effective and safe in terms of treating chronic progressive moderate and severe IgA nephropathy with hyperuricemia.

Objective to measure the percentage of th22 cells and evaluate the levels of plasma interlukin-22(IL-22)in human peripheral blood, so as to determine their clinical significance in primary Sj?gren′s syndrome(pSS). Methods Patients with pSS were divided into three subgroups based on severity of labial gland involvement:mild,moderate and severe. Healthy people served as controls. the percentage of th22 cells and the lev-els of IL-22 in human peripheral blood from pSS patients and healthy controls were measured and compared. Results Compared to healthy con-trols,pSS patients had significantly higher percentage of th22 cells and higher plasma IL-22 levels(P < 0.05). Among pSS patients,the more seri-ous illness they had,the higher percentage of th22 cell and levels of IL-22 were observed. Severe patients had higher percentage of th22 cells and IL-22 levels than moderate patients(P < 0.05),and moderate patients had higher percentage of th22 cells and IL-22 levels than mild patients(P <0.05). Conclusion Increased peripheral IL-22-secreting th22 cells are detected in primary Sj?gren′s syndrome,which have a close association with disease severity. these data suggest that th22 and its released cytokine IL-22 may be considered as potential valuable biomarkers for severity of pSS,which may provide a novel therapeutic target for treatment.

AIM: To prepare a soluble hemangiopoietin(HAPO) protein and to construct pET22b(+) expression vector, to obtain pure recombinant HAPO protein and to measure its bioactivity. METHODS: HAPO cDNA was amplified using RT-PCR method from a commercial human fetal liver cDNA library. The resulting product was cloned into pET22b(+) vector and transformed into E.coli BL21(DE3). The recombinant protein was isolated and purified by Ni~(2+)-NTA chelating resin and the chromatographies of SP Sepharose FF. The adhesion of human umbilical vein endothelial cells (HUVEC) were measured by adhesion assay. RESULTS: HAPO gene with a reading frame of 897 bp was successfully cloned from human fetal liver cDNA library, the expressed pET22b(+)-HAPO fused protein existed in a soluble form, with the yield above 10% total bacterial protein and its purity achieved above 80%. The activity assay showed that the treatment of HAPO enhanced total adherence of HUVEC in a concentration-dependent manner. CONCLUSION: HAPO protein can be expressed in a soluble form. HAPO may facilitate the homing of hematopoietic stem/progenitor cells in vitro.

Objective The purpose of the experiment is to research the effects of different kind of anti-platelet drugs and their combinations in prevention of vascular restenosis after ballon-injury, and to evaluate the best way of clinical utility of anti-platelet drugs.Methods Fifty-six male New Zealand White Rabbits were randomly assigned to 3 groups: the control group (ML) , the model group (M) , the experimental group.The experimental group was divided to 5 subgroups, the aspirin group (A), the cilostazol group (B) , the clopidogrel group (C), the aspirin plus cilostazol group (A + B) , and the aspirin plus colopidogrel group (A + C).Femoral artery stenosis model was induced by halloon injury except the control group.The rabbits were fed different drugs 3 days before operation except the normal group and the model group.On the 14th day after operation, the rabbits were sacrificed and the iliofemoral arteries were removed and made to the pathological section.We measured the internal elastic lamina, external elastic lamiua and luminal areas measured.Then calculate the area of intima, media and the ratio of intima vs.media.Results ①There is no endothelium hyperplasia in control group.Compared with the control group, we observed endothelium hyperplasia in both model group and the experimental groups.Compared with the model group, endothelium hyperplasia in all experimental groups is in a lower degree;② Cilostazol alone works better than aspirin alone and clopidogrel alone in prevention of vascular restenosis after ballon-injury.Clopidogrel combined with aspirin and cilostazol combined with aspirin both work better than aspirin alone in prevention of vascular restenosis after ballon-injury.There is no apparent difference between the effect of clopidogrel combined with aspirin and cilostazol combined with aspirin.Conclusions ① Different anti-platelet drugs and different combinations of them can prevent the happening of vascular restenosis after ballon-injury.② Cilostazol works better than aspirin and clopidogrel in prevention the happening of vascular restenosis after ballon-injury, and combination of anti-platelet drugs which contains cilostazol works better than other way of combination of anti-platelet drugs.③ Cilostazol and combination of anti-platelet drugs which contains cilostazol is potential useful in clinical uses to prevent restenosis after PTA.

Objective: To compare regeneration of motor fibers with that of sensory fibers by end to side neurorrhaphy and compare the effect of end to side neurorrhaphy with that of end to end neurorrhaphy. Methods: 20 SD rats were randomly divided into two groups: group A and group B. In group A, the right peroneal nerve was sectioned and the distal end was sutured laterally to the tibial nerve; in group B, the right peroneal nerve was sectioned and sutured with end to end neurorrhaphy. All left sides of two groups were used as control. Retrograde transportation of HRP was observed after 28 weeks. Results: The labelled neurons were also found in the anterior horn of the spinal cord and the spinal ganglia. Conclusion: The regenerative nerve fibers had motor fibers as well as sensory fibers by end to side neurorrhaphy. The effect of end to side neurorrhaphy is not as good as that of end to end neurorrhaphy. [

Objective: To investigate the expression of midkine (MK) gene in childhood acute lymphoblastic leukemia (ALL) and the clinical significance of MK thereof. Methods: The real-time PCR was used to assay MK gene expression in bone marrow of 15 normal children and 124 childhood ALL patients, including 73 patients in progression and 51 patients in complete remission. Three stratifications of progressing patients were established by prognostic factors such as white blood cell count, age, immunopherotype and response to the 7-day prednisolone prephase. Results: The significant statistic difference in MK gene expression was found between the progression group, the complete remission group and the normal group (P< 0.01). The MK gene expression was over-expressed in B-ALL than that in normal group. Furthermore, there was statistic difference between B-ALL and T-ALL (P< 0.01). But there was no difference in MK mRNA expression between the normal control and T-ALL. The assay in risk stratifications showed that the levels of MK gene were higher in standard risk group and mid-risk group than that in high risk group (P< 0.01 and P< 0.05, respectively). There was no significant difference between standard risk group and mid -risk group (P = 0.32). No correlations were found between MK level and age, gender or lactate dehydrogenase level in serum. The expression of MK was significantly lower in the group with higher white blood cells(WBC≥ 25×10~9/L) than that with lower WBC (WBC<25×l0~9/L) in peripheral blood (P< 0.05). Conclusion: The high level of MK was a favorable prognostic factor in childhood acute lymphoblastic leukemia patients.

BACKGROUND; As the bridge vessel, radial artery is easy to occur spasm and injury of endometrium, and the abnormal proliferation of endometrium, re-vasospasm, etc. appear at early period postoperatively, which can affect the operative outcomes.OBJECTIVE: To compare the effects of nitroglycenn (NTG), verapamil (VP), paraverine (PA), and the mixture of NTG and VP on relaxing spasm of radial artery in human being.DESIGN: A randomized controlled trial.SETTING: Department of Cardiosurgery, General Hospital of Shenyang Military Area Command of Chinese PLA.PARTICIPANTS: Thirty patients who underwent coronary artery bypass graft (CABG) with autologous radial artery, were selected from the Department of Cardiosurgery, General Hospital of Shenyang Military Area Command of Chinese PLA from September to November in 2006, including 18 males and 12 females, 48-74 years of age. Informed consents were obtained from all the patients or their relatives preoperatively.METHODS:① The samples of human radial artery in CABG were collected, and totally 95 vascular rings were used in this study. The rings were divided into five groups: NTG group, VP group, PA group, VP+NTG group and control group.There two parts of the experiment, vasospasm relaxation test and vasospasm prevention test. The vascular rings in the control group were only immersed in the Krebs- Ringer solution without any drug. ② Drugs: NTG was purchased from Solo Pak Laboratories (Franklin Park, IL), PA from Eli Lilly & Co., (Indianapolis, IN), VP and others from Sigma (St.Louis, MO). ③ Thirty vascular rings were used in the vasospasm relaxation test, and the method of bath chamber was applied. Deoxyepinephrine (terminal concentration of 1 ×10-3 mol/L) was added to stimulate the contraction and spasm of the vascular rings, then four vasodilatators were added, the concentrations of VP and NTG were 30 μmol/L, and that of PA was 0.1%. Relaxation rate=(resting tension after stimulation-initial resting tension)/initial resting tension× 100%. ④ The other 60 vascular rings were used in the vasospasm prevention test. Firstly, the rings were immersed in the four solutions of vasodilatators of corresponding concentrations and Krebs- Ringer solution respectively for 30 minutes. After pretreatment, the vascular rings were randomly divided intc normothermia group (n =30) and cryopreservation group (n =30). In the cryopreservation group, the vascular rings were put into the Krebs-Ringer buffer solution at 4 ℃, and kept in refrigerator at 4 ℃ for 24 hours. The spasms of radial artery were observed.MAIN OUTCOME MEASURES: Changes of resting tension (degree of contraction and relaxation rate of dilatation) of vascular rings before and after administration.RESULTS: ① Effect on the dilatation of spastic radial artery: The radial artery was completely within 11 minutes in all the VP+NTG group, VP group, NTG group and PA group. But in the first 3 minutes, the dilating effects of VP+NTG and NTG were obviously better than the other two groups. The dilatation curves showed that the dilating ability in order was VP+NTG ＞ NTG ＞ VP ＞ PA. ② Effect of pretreatment of radial artery on antispasm: In the normothermia group, the contractility of the vascular ring produced by 1 ×10-3 mol/L deoxyepinephrine was close between the VP+NTG group and VP group [(0.47±0.06), (0.49±0.08) g, P ＞ 0.05], which were obviously different from those in the NTG group and PA group [(0.81±0.22), (0.87±0.26) g, P ＜ 0.05]. After cryopreservation for 24 hours, the contractility in the VP+NTG group was not obviously different from that in the VP group [(0.86±0.11), (0.90±0.13) g, P ＞ 0.05], and obviously lower than those in the NTG group, PA group and control group [(4.82±0.87), (5.00±0.53), (5.10±0.67) g, P ＜ 0.01], whereas those in the NTG group and PA group were close to that in the control group (P ＞ 0.05).CONCLUSION: The drugs can prevent and relax the spasm of radial artery to different extents, whereas considering from preventing spasm and the time-effect of treatment, the mixture of VP and NTG seems to be more proper as the preparation solution for the treatment of radial artery in CABG.