Objective To investigate the molecular basis of ABO gene in a patient with serologic ABO blood group discrepancy.Methods Serologic blood group identification,Coombs' test and antibody screening were detected with DG Gel Confirm cards,Neutral cards,Coombs cards by WADiana/8XT Compact Analyzer (from Diagnostic Grifols,S.A).The enhancer,promoter,exon 1 ～ 7 and their adjacent intron region of ABO gene were amplified by using polymerase chain reaction (PCR) method.Results The patient's red blood cells was determined as weak B phenotype showing two groups in gel and mixed field in tube with monoclonal anti-B,and A phenotype with monoclonal anti-A.DNA sequencing showed nine variants in ABO gene.One heterozygous variation in exon 6 (297A＞G) and eight heterozygous variations in exon 7 (467C＞T,526C ＞G,657C＞T,703G＞A,796C＞A,803G＞C,829G＞T 930G＞A) were identified and 829G＞T was the novel variant.Compared with Blood Group Antigen Gene Mutation Database,genotype of the patient was weak expression of A102/B101.Conclusion The novel variation of B allele is the main reason of Bweak phonetype in A102/B101 genotype.Serological and molecular biological detection help to understand the characteristics of blood group phenotype and genotype,provide the guidance for clinical transfusion strategies.

Objeetive To explore the differentiation of B lymphocytes and expression of B7-related protein-1 (B7RP-1)on B lymphocytes in patients with systemic lupus erythematosus(SLE).Methods Three-color immunofluorescent staining and flow cytometric assay were used to analyze the frequency of three types of B lymphocytes,I.e.,plasma cells,memory B lyphocytes and naive B lymphocytes,as well as the expression of B7RP-1 on these cells in peripheral blood from 23 patients with SLE and 16 normal human controls.Clinical data of these patients with SLE were collected.and SLE disease activi index(SLEDAI)was also evaluated.The relationship was assessed between the expression of B7RP-1 and SLEDAI.Results The frequency of plasma cells was highest in patients with active SLE.followed by patients with inactive SLE and normal human controls(P＜0.01).A significant decrease was observed in the frequency of memory B lymphocytes in patients with active SLE compared with normal controls (P＜0.01),but no significant difference was found between patients with inactive SLE and those with active SLE(P＞0.05).Regarding the frequency of naive B lymphocytes,there was no significant difierence among the three groups.Increased frequency of plasma cells was also noted in patients with lupus nephritis(LN)compared with those without LN [(6.15±3.12)%vs(3.31±1.41)%,P＜0.05 ],but no significant difierence was found with regard to the frequency of memory or naive B lymphocytes between these two groups.The expression rate Of B7RP-1 was significantly lower on total lymphocytes from patients with SLE than from normal human controls (46.51%vs 63.75%,P＜0.05),which was the case with B7RP-1 on plasma cells,memory B lyphocytes and naive B lymphocytes (all P＜0.01),whereas no significant difierence was found between patients with inactive SLE and active SLE or between patients with and without LN.In addition.no correlation was found between the expression of B7RP-1 and SLEDAI(r=0.035,P＞0.05).Conclusions In peripheral blood of patients with SLE,the frequency of plasma cells is increased,while the expression of B7RP-1 on lymphocytes is decreased,which may be relevant to the pathogenesis of SLE.

Objective To assess the diagnostic value of MRI on multiple focal nodular hyperplasia (FNH) of the liver. Methods MR images of 9 cases with pathological-confirmed multiple FNH were retrospectively analyzed. MRI features of the lesions were correlated with pathological findings. Results Multiple FNH was considered in all these 9 cases. Among them, the primary diagnosis was FNH in 5,hepatic adenoma in 3 and fibrolamellar hepatocellular carcinoma in 1 case. A total of 31 lesions were detected in the 9 cases. On T2WI, 19 lesions presented slightly high-signal intensity, and the other 12 presented iso-signal intensity. On T1WI, 12 lesions presented slightly low-signal intensity, 7 presented iso-signal intensity, and the other 12 presented high-signal intensity. On opposed-phase, the signal intensity of 1 lesion dropped unevenly. After bolus injection of contrast agent Gd-DTPA, in hepatic arterial phase 18 lesions showed mild to marked heterogeneous enhancement, 11 showed marked homogeneous enhancement, 1 showed moderate ring-like enhancement, and the last one did not have obvious enhancement In portal venous and delayed phase, all the lesions turned to iso- or slightly high-signal intensity gradually. Sixteen of 31 lesions presented central scar, which demonstrated mild star-like enhancement in delayed phase. Conclusion Multiple FNH presented certain MRI features, which contributed to the preoperative diagnosis.

OBJECTIVETo study the correlation between EEG characteristics of medial prefrontal cortex (mPFC) and drug-seeking behavior of rats with morphine dependent place preference under shuttling condition.

METHODSForty rats were randomly divided into four groups (n = 10): morphine PL group, NS PL group, morphine IL group and NS IL group. After embeding the electrode in prelimbic (PL) or infralimbic (IL) cortex of each group by brain stereotaxic operation, the model of morphine dependent conditioned place preference (CPP) in rats was established. The differences of EEG wave percentage in mPFC were telemetered and analyzed when rats shuttled before and after the model.

RESULTSAfter the model, the withdrawal symptoms were evident in morphine PL and IL group, and the activity time and distance in white box were increased obviously. Compared with control group, after the model, the EEG in morphine PL group showed that: when the rats shuttled to white box, 8 wave decreased obviously, P wave increased obviously. When the rats shuttled to black box, brain waves showed opposite changes. The EEG in morphine IL group showed that: when the rats shuttled to white box, a wave increased obviously, P and a wave decreased obviously. When the rats shuttled to black box, the brain wave had no significant differences compared with control group.

CONCLUSIONThe EEG changes are different in PL and IL cortex of morphine CPP rats under shuttling condition, and the EEG changes are also different when rats shuttling to white or black box. There is possibly different mechanism, when different drug-seeking environmental cues caused EEG changes in different regions of mPFC.

Animals ; Conditioning (Psychology) ; Cues ; Drug-Seeking Behavior ; Electroencephalography ; Morphine Dependence ; physiopathology ; Prefrontal Cortex ; physiopathology ; Rats ; Telemetry

OBJECTIVETo analyse the relationship between the electrical activity changes of nucleus accumbens (NAc) shell and the drug-seeking behavior by recording NAc shell electrical activity in conditioned place preference (CPP) rats induced by morphine.

METHODSForty SD rats were randomly divided into operation-only control group and the morphine-induced CPP group after stereotaxic electrode was buried on rats NAc shell and the latter group was used to establish the morphine CPP model(n = 20). A CPP video system combining with the technique of electrical activity wireless telemetry was used in the study. The NAc electrical activity from each group of rats was recorded by wireless telemetry respectively, which included staying in black or white chamber of video box, shuttling between black-white chambers and between white-black chambers. The electrical activity differences were analyzed by the percentage of each wave.

RESULTSWhen the morphine-induced rats staying in black chamber, compared with the operation-only control group, the NAc shell electrical activity showed that the percentage of 0 - 10 Hz was increased(P < 0.05), meanwhile, those of 10 - 20 Hz and 30 - 40 Hz were reduced(P < 0.05, P < 0.01); when the morphine-induced rats staying in white chamber, the NAc shell electrical activity showed that the percentage of 0 - 10 Hz and 30 - 40 Hz were increased(P < 0.05 , P < 0.01) , that of 10 - 20 Hz was reduced(P < 0.05 , P < 0. 01); when the morphine-induced rats in black- white shuttling status, the NAc shell electrical activity showed that the percentage of 0 - 10 Hz was increased(P <0.05, P <0.01), that of 10- 30 Hz was reduced( P <0.05); and in the white-black shuttling status, the electrical activity showed that the percentage of 0 - 10 Hz was reduced(P <0.05), that of 10 - 30 Hz was increased(P < 0.05) ; the electrical activity was further compared between staying status and shuttling status in the morphine-induced CPP group. There was no significant difference of electrical activity between the rats in white-black shuttling status and staying in white chamber. However, when rats in black-white shuttling status, compared with staying in black chamber, the electrical activity showed that the percentage of 0 - 10 Hz and 40 - 50 Hz were increased(P < 0.05), meanwhile, those of 10 - 20 Hz and 30 - 40 Hz were reduced(P <0.05).

CONCLUSIONThe electrical activity changes of NAc shell in morphine-induced CPP rats were different from those of the operation-only control group, and these changes might be associated to the rat's drug-seeking behavior.

Animals ; Conditioning (Psychology) ; Drug-Seeking Behavior ; Morphine ; pharmacology ; Nucleus Accumbens ; drug effects ; physiology ; Rats ; Rats, Sprague-Dawley ; Telemetry

It is generally accepted that tissue factor plays an important role in coagulation and intravascular thrombus formation. Tissue factor is not only found primarily on the surface of certain cells that are located outside the vasculature, but also found in circulating cells. Monocyte express tissue factor induced by endotoxin. Recently, many researches indicate that P-selectin, CD40 ligand and GPIIb/IIIa receptor of platelet can also affect expression of tissue factor by monocyters. In addition, a lot of studies showed that tissue factor exist in the circulation including contained platelet. Tissue factor in the platelet releases under certain condition, and initiates coagulation. In this review the relation between platelet and tissue factor was elaborated.
Blood Platelets ; drug effects ; metabolism ; physiology ; CD40 Ligand ; physiology ; Humans ; Monocytes ; drug effects ; metabolism ; P-Selectin ; physiology ; Peptides ; pharmacology ; Platelet Glycoprotein GPIIb-IIIa Complex ; physiology ; Thromboplastin ; biosynthesis ; drug effects ; physiology

Objective To investigate the relationship between activation of gliacytes , mitogen-activated protein kinase (p38MAPK) and neuronal apoptosis after microinjecting aggregated Aβ25-35 into hippocampus.Methods The model was established by using stereotaxic technique to inject 10μg aggregated Aβ25-35 into dorsal hippocampus in rats .The rats were grouped as the control , vehicle and model groups .Immunohistochemistry and Western blotting were used for detection of activation of microglia(MG), atrocytes (AS) and expression of p-p38MAPK in the hippocampus.ELISA was used to evaluate the level of TNF-αand IL-1β.The survival neurons were observed by Nissl staining and the apoptotic neurons were identified by tunnel staining .Results Expression of ox-42, GFAP, p-p38MAPK were up-regulated in hippocampus, as well as TNF-α、IL-1β, which reached a highest value on the 7th day after injection of Aβ25-35.However, the number of neuron with Nissl positive decreased gradually , and the tunnel positive neurons increased highly and reached a peak value on the 7th day.There were significant differences between the control and vehicle group ( P <0.01). Conclusion Apoptosis of the neuron caused by Aβ25-35 injection may result from activation of gliacytes , p38 MAPK and increase of TNF-αand IL-1βlevel.

Objective To evaluate the outcomes of mature B-cell acute lymphoblastic leukemia(mature B-ALL) and to assess the safety and efficacy of the treatment protocol.Methods From February of 2003 to December of 2012,15 children were diagnosed as mature B-cell acute lymphoblastic leukemia/lymphoma possible (mature B-ALL/NHLp) in Shanghai Children's Medical Center(SCMC) were enrolled,and they were treated with SCMC-mature B-ALL/NHLp-2003 protocol.All of the clinical characteristics,therapeutic effects and long-term outcomes were analyzed.The statistical data were processed by SPSS 21.0.Results The median age on diagnosis was 8.7 years (1 year and 5 months to 14 years and 4 months).Among them,4 cases presented with local mass including maxillofacial tumors,neck and abdominal mass.The others had systemic manifestations such as fever and pale face.These neoplastic cells retained the expressions of surface membrane immunoglobulin M,terminal deoxynucleotidyl transferase,Cμ,CD10,CD19,cCD79 a differently.Follow-up was updated to November 30,2013.The median follow-up period was 80 months (39-128 months).Theestimated 5-year event free survival rate was (80.0 ± 10.3) ％.According to univariate analysis,increased lactate dehydrogenase level (＞ 4-times the normal value),increased serum ferritin level (＞ 2-times the normal value),no small residual disease markers were indepen-dent poor prognostic factors(x2 =5.49,4.89,5.49,all P ＜ 0.05).Conclusions SCMC-mature B-NHL/ALLp-2003 protocol is feasible and safe for children with mature B-ALL/NHLp,but more sample cases need to be investigated.

OBJECTIVE:To study the intervention mechanism of puerarin on expressions of HSP70 and Fas proteinum in rats with acute cerebral ischemia injury.METHODS:12 rats with acute cerebral ischemia injury were randomly divided into ischemia control group(saline) and puerarin treated control group,with 6 in each group,each rat under took own control by ischemia side(exp.group) and non-ischemia side(control group);And rats with acute cerebral ischemia injury were randomly divided into control group (saline,ischemia side),puerarin treated control group and normal group (non-ischemia side),with 6 in each group.And then the expressions of HSP70 and Fas proteinum is determined by HE staining and immunochistochemistry SP.RESULTS: The expression of HSP70 was negative or weakly positive in ischemia control group and puerarin treated control group, while that in ischemia experiment group and puerarin treated experiment group was significantly increased(P0.05);The numbers of dead cells in control group and puerarin group were found to be significantly larger than that in normal group(P

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