This paper investigate the methods to detect engraftment rate of the four patients with hematological disorders who accepted nonmyeloablative allogeneic peripheral blood stem cell transplantation(NAPBSCT). To find out the best method, their engraftment rates were detected serially at different time after NAPBSCT by means of either FISH, or conventional chromosome analysis combined with R banding analysis concurrently.The results were carefully compared with one another. All these four sex mismatched cases were engrafted partially,and two of them changed to full engrafment. The results show no statistically significant difference in 3 groups (conventional method, FISH for hypermetaphase, FISH for interphase nuclei). But the results strongly indicate that FISH is a rapid, precise, objective,and reliable menthod for detection of the engraftment rate,and it is suitable for sex mismatched NAPBSCT.

This paper investigate the methods to detect engraftment rate of the four patients with hematological disorders who accepted nonmyeloablative allogeneic peripheral blood stem cell transplantation(NAPBSCT). To find out the best method, their engraftment rates were detected serially at different time after NAPBSCT by means of either FISH, or conventional chromosome analysis combined with R-banding analysis concurrently.The results were carefully compared with one another. All these four sex-mismatched cases were engrafted partially,and two of them changed to full engrafment. The results show no statistically significant difference in 3 groups (conventional method, FISH for hypermetaphase, FISH for interphase nuclei). But the results strongly indicate that FISH is a rapid, precise, objective,and reliable menthod for detection of the engraftment rate,and it is suitable for sex-mismatched NAPBSCT.

Objective To investigate the effect of lipoxin A4 ( LXA4 ) on the permeability of blood-brain barrier (BBB) after focal cerebral ischemia-repeffnsion (I/R) in rats. Methods Fifty-four adult male SD rats weighing 200-250 g were randomly divided into 3 groups ( n = 18 each): group Ⅰ sham operation (group S); group Ⅱ focal cerebral I/R ( group I/R) and group Ⅲ LXA4 ( group L). Focal cerebral I/R was produced by middle cerebral artery occlusion (MCAO) with a 4-0 nylon thread with rounded tip inserted into right internal jugular vein and threaded cranially in group Ⅱ and Ⅲ . In group Ⅲ LXA4 100 ng was injected into right lateral ventricle of the brain after MCA was successfully occluded. MCAO was maintained for 2 h. The neurological deficit was evaluated and scored (0 = no deficit, 5 = death) at 24 h of reperfusion. 2% Evans blue 4 ml/kg was injected via femoral vein at 1 h before the animals were sacrificed. The animals were killed and their brains were immediately removed for determination of brain water content, Evans blue content and expression of matrix metalloproteinase-9 (MMP-9)in the ischemic cortex. Results The neurologic deficit scores, the brain water and Evans blue content and MMP-9 protein expression in the cortex were significantly higher in I/R group than in S group. The cerebral I/R-induced changes were significantly attenuated in LXA4 group. Conclusion LXA4 can protect blood-brain barrier against cerebral I/R injury by inhibiting MMP-9 protein expression in the brain tissue.

Objective To investigate the value of cytokinesis-block micronuclei(CBMN)assay in estimation of the biological doses of the victims of radiation accident.Methods Samples of peripheral blood were collected from the 5 victims(Subjects 1-5)at 16 h after the radiation accident of Taiyuan,Shanxi Province.And the peripheral blood samples and bone marrow sample were collected from the victim No.1 at 23 and 24 h after the radiation.Eight days after the accident Subject 1 underwent allogeneic peripheral blood hematopoietic stem cell transplantation.At difierent time points in the period of 1 year after the accident.peripheral blood samples were collected from these 5 victims.CBMN assay was conducted on the peripheral blood lymphocytes on the samples,and the biological doses were estimated based on the micronuclei(MN)frequencies.The nuclear division index(NDI)obtained from in vitro irradiation experiment using high dose of 60Coγ-rays was used to estimate the exposed doses for Subject 1. Dynamic arialysis of the MN frequency for the 5 victims was performed in the period of 1 year after the accident.Results The MN frequency of Subject 1 surpassed the value corresponding to the upper limit of the MN dose.effective curve.The dose range estimated bv the combination of the CBMN and NDI (CBMN+NDl)assay was 10-20 Gy for Subject 1.The doses estimated by MN frequency for Subjects 2,3,4,and 5 were 3.6,2.9,2.3,and 2.9 Gy,respectively.The estimated doses were in accordance with those estimated by physicat method.chromosome aberration analysis.and clinical symptoms.Prominent decrease of the MN frequency was observed at 26 d after the accident(18 d after the transplantation)for Subject 1(u=3.295,P<0.05).Gradual decrease of MN frequency was observed after the accident for Subjects 2,3,4,and 5.The MN frequencies 1 month after the accident of Subjects 3,4,and 5 were all significantly lower than those 16 h later(u=6.874,4.526,and 7.811,P<0.05).Conclusions Quick and accurate.CBMN assay reinforces and verifies the result of chromosome aberration analysis.The new index CBMN+NDI assay is of reference valne for estimating higher dose of irradiation.

Objective To investigate the effect of lipoxin A4(LXA4) on inflammatory response in a rat model of permanent focal cerebral ischemia (PFCI). Methods Seventy-two adult male SD rats weighing 200-250 g were randomly divided into 3 groups (n = 24 each):group Ⅰ sham operation (group S); group Ⅱ PFCI and group Ⅲ LXA4. PFCI was induced by thread occlusion of right middle cerebral artery according to the method described by Longa in group Ⅱ and Ⅲ. In group Ⅲ LXA4 100 ng/5 μl was injected into right ventricle of the brain after PFCI was successfully induced, while in group Ⅰ and Ⅱ equal volume of normal saline was injected instead of LXA4. Six animals were killed at 6, 12 and 24 h of ischemia. Their brains were immediately removed for microscopic examination and determination of myeloperoxidase (MPO) activity, TNF-α, IL-10 and transforming growth factor-β1 (TGF-β1) contents in the ischemic cortex. The expression of glial fibrillary acidic protein (GFAP)was measured by immuno-histochemistry. Apoptosis in neurons was assessed using TUNEL. Results PFCI significantly increased MPO activity, TNF-α, IL-10 and TGF-β1 contents and GFAP expression in the ischemic cortex and neuronal apoptosis in group Ⅱ as compared with group S. LXA4 significanfly decreased MPO activity,TNF-α content, GFAP expression and neuronal apoptosis and increased IL-10, TGF-β1 contents at 12,24 h of ischemia. LXA4 significantly ameliorated PFCI-induced cerebral histopathologic damage. Conclusion LXA4 can protect the brain against PFCI injury by inhibiting inflammatory response.

Objective To establish a dose-effect curve of premature condensation chromosome ring (PCC-R)in lymphocytes of human peripheral blood after exposed to high doses of Y-rays. Methods Peripheral blood samples was drawn from three healthy individuals, and exposed to 60Co γ-rays with doses between 0 and 30 Gy. The frequencies of PCC-R in premature condensation chromosome (PCC) cells obtained by Okadaic acid (OA) induction were calculated, and a dose-effect curve was fitted. Results PCC index tapered with dose. Frequencies of PCC-R per cell increased until 20 Gy, and then saturation was observed. The results were fitted to a lineal model up to 20 Gy: y = - 0.020 + 0.052D ,where y was the frequencies of PCC-R per cell, D was the radiation dose (Gy). Conclusins The highest dose could be estimated is 20 Gy by the dose-effect curve established with PCC-R method. Its utility and validity will be verified in the future application of radiation accident.

Objective To investigate the effect of the asthmatic mice's contents of NF-κB、IκB、p-IκB treated with curcumin.Methods Thirty Balb/c mice were divided into three groups randomly,the normal mice,the asthmatic mice and the curcumin mice.The bronchial hyperresponsiveness of the three groups were exanined by lung function.The NF-κB 、IκB 、p-IκB content of three teams were tested.Results The content of cytoplasm NF-κB in asthmatic mice was lower than the control (P ＜ 0.01).However,the content of cytoplasm NF-κB in curcumin team was more than the asthmatic mice (P ＜ 0.05).In the other hand,The content of nuclear NF-κB in asthmatic mice was more than the control(P ＜0.01).However,the content of nuclear NF-κB in curcumin mice was lower than the asthmatic mice (P ＜ 0.05).The content of cytoplasm p-hκB in asthmatic mice was more than the control and the content of IκB was lower than the control(P ＜ 0.01).Howerer,the content of IκB in curcumin team have much more than the asthmatic mice and the content of p-IκB have much lower than the control(P ＜ 0.01).Condusion Curcumin alleviates the airway hyperresponsiveness of the asthmatic mice through the suppression of NF-κB transcribe to the nuclear via alleviating the phosphorylation of I-κB.

Objective To investigate the effect of the asthmatic mice's contents of nuclear factorerythroid-2-related factor 2,Nrf2 and HO-1 reated with curcumin in lung tissue.Methods All 45 mice were divided into three teams randomly,the normal mice,the asthmatic mice and the curcumin mice group.We tested the expression of Nrf2 and HO-1 of three mice teams in lung by immunolfluorescence technique.We tested the protein contents of Nrf2 and HO-1 of the three teams in the lung tissue by Western blot.We tested the binding activity of Nrff2 and ARE of the three teams in lung by EMSA.Results The expression of Nrf2 and HO-1 in curcumin group was significantly higher than that in asthma group and normal control group by immunolfluorescence technique.There was no sigrnificant difference in content of cytoplasm Nrf2 protein in lung tissue between three groups by western blot(P ＞ 0.05).The content of nuclear Nrf2 and HO-1 protein in the lung tissue of the curcumin group was significantly higher than that in the other two groups,the difference is statistically significant (Nrf2,P ＜0.05;HO-1,P ＜0.01).The binding activity of Nrf2-ARE in lung tissue of curcumin group was significantly higher than that of asthma group and control group,with statistical significance(P ＜ 0.05).Conclusion Curcumin can increase the protein contents of the Nrf2 and HO-1 of mice and enhance the expression of them.

Objective To understand the correlation of gene detection of Mycoplasma pneumoniae and clinical refractory Mycoplasma pneumoniae pneumonia.Methods (1) For children with Mycoplasma pneumoniae pneumonia in our hospital with serum Mycoplasma pneumoniae antibody positive,we collected the pharyngeal swab specimens over the same period,applied nested PCR to amplify 23SrRNA gene and undergoing electrophoresis and find out 97 cases of both positive,conducted DNA sequencing analysis of macrolide resistant gene to isolate the mutants,compared clinical manifestations of drug-resistance gene group with no drug-resistance gene mutation group.(2) Ninety-seven cases of mycoplasma pneumonia (MPP) patients were devided into the general MPP group (68 cases) and refractory MPP group (29 cases),retrospectively analyzed clinical manifestation,laboratory examination and differences of imaging performance.Multivariate logistic regression analysis for the performance of refractory Mycoplasma pneumoniae pneumonia was carried out to examine whether there is relevance between the mutant of drug-resisting gene and refractory Mycoplasma pneumonia.Results (1) Seventeen of 97 cases (17.5％) were found out without mutations,the other 80 cases (82.5％) exist drug-resistance gene mutations.(2) Mutation of drug-resistance gene group showed high CRP values,heating time,hospitalization time,macrolide drug application time,application of macrolides fever time and longer cough,by statistical analysis with statistical significance,higher incidence of lobar pneumonia.(3) Compared to general MPP group,refractory MPP group showed high peripheral blood neutrophil percentage percentage,CRP,calcitonin) and lactate dehydrogenase (LDH) values,heating time,hospitalization time,macrolide drug application time,application of macrolides fever time and longer cough.There was significant difference (P ＜ 0.05);macrocyclic lactones drug application time and resistance gene mutation and refractory Mycoplasma pneumonia were correlated.Conclusion MPP drug-resistant genes are widespread.Drug resistance gene mutations group shows long clinical symptoms duration,slow recovery rate,higher CRP value,higher rates of lobar pneumonia.Compared with ordinary MPP group,there are higher drug resistance mutation rate,inflammatory indexes and lactate dehydrogenase value,large ring lactone class drugs after a longer time of cough and fever in RMPP group.Drug application time and resistant gene mutations are associated with RMPP.

Objective To investigate the detection and clinical features of primary immunodeficiency disorders (PID) in children for an earlier diagnosis of disease.Methods The clinical data of the 27 children with PID diagnosed in Shengjing Hospital Affiliated to China Medical University from Dec.2003 to Nov.2011 were reviewed,including illness history,birth history,family history,clinical feature,laboratory data,diagnosis,treatment and outcome,etc.Results In 27 children with PID,antibody deficiencies were the most frequent findings (48.15％,13/27 cases),followed by combined immunodeficiency (22.22％,6/27 cases),phagocytic disorders (14.81％,4/27 cases),and immunodeficiency with other major defects accounted for 14.81％ (4/27 cases).PID was characterized by recurrent,severe and prolonged infection,but all kinds of PID had their own clinical features.Recurrent infections occurred in 24 cases.Respiratory infections and otitis media were the most common clinical manifestation.Seven patients had a family history.The fatality rate was 37.04％ (10/27 cases).Conclusions There are vast varieties of PID in our area and antibody deficiencies are the most common type.All kinds of PID have their own clinical features,which may guide us to choose appropriate lab examination.There are nearly 25％ patients with PID who have family history.The fatality rate is high.Patients who suffer from recurrent infections,especially respiratory infections or otitis media,or those with a family history should have early immunology testing so as to be detected and diagnosed of PID earlier.