Objective To investigate the effects of macmphage migration inhibitory factor (MIF) on glucocorticoid (GC) re]ease and glucocorticoid recer (GR) in mts.Methods Test Ⅰ Thirty-two male SD rats weighing 250-300 g were randomly divided into 4 groups(n=8 each):control group(C),low dose recombinant MIF (rMIF) group (rMIF-L),middle dose rMIF group (rMIF-M) and high dose rMIF group (rMIF-H).The animals received l ml normal saline via the right femoral vein in group C.The animals received rMIF50.100 and 200 ng in l ml normal saline though right femoral vein in group rMIF-L,rMIF-M or rMIF-H respectively.Blood samples were taken from left femoral artery immediately before inection(T0,baseline),and at 5 min,3 h,6 h.12 h and 24 h after injection of rMIF(T1-5) for determination of serum concentration of corticosterone.Test Ⅱ Primary cultured neonate rat(2-3 days)myocardial ceils were randomly divided into 3 groups(n=24 each):group C,group rMIF-L and group rMIF-M.The ceUs in group C,rMIF-L and rMIF-M wefe incubated with DMEM.rMIF 50 ng+DMEM and rMIF 100 ng+DMEM for 3 h respectively.The expression of GR and HsPg0 wag determined by Western blot.ResuBs Test Ⅰ The serum concentration of corticosterone was signifieemily higher in the other 3 groups than in group C at T1-5(P<0.05).The sertlm concentration of corticostemne was significantly increased at T1-5 in group rMIF-L,rMIF-M and rMIF-H compared with the baseline values(P<0.05).Test Ⅱ HSP90 expresion was significantly lower in the other two groups than in group C(P<0.05).Them was rio signifieanf difference in HSP90 expression between group rMIF-L and group rMIF-M(P>0.05).There was no significant difference in GR expression among the 3 groups ( P > 0.05). Conclusion MIF druing sepsis can weaken GR function through down-regulating HSP9O expression, resulting in CC resistance.

Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; metabolism ; Proto-Oncogene Proteins c-akt ; metabolism ; Signal Transduction

Objective To investigate the role of tyrosine kinase Src in a murine C.albicans infection model. Methods Observed cell proliferation by alarmarblue assay at 2, 24 and 48 h after Src inhibitor PP2 treatment. Phagocytosis was determined by a fluorometric assay. Cytokine TNF-αand IL-10 production was detected by ELISA. Results The 0~33.3 μmol/L PP2 had no effect on cell proliferation after PP2 treatment for 2 h. When the PP2 treatment extended to 24 or 48 h, PP2 (11.1, 33.3μmol/L) showed significant inhibition on cell proliferation with 78%, 9%, and 54%,13%, respectively. At 48 h after 11.1μmol/L PP2 treatment, the internalization of C.albicans in macrophage is significantly inhibited, contributing to the inhibition of cell proliferation. However, the 11.1 and 33.3μmol/L PP2 significantly inhibited the cytokine TNF-αand IL-10 production during C.albicans infection (P<0.01). Conclusion Src kinase played an important role during C.albicans infection, especially for the cytokine TNF-αand IL-10 production.

Objective: We have studied the role of lymphoproliferative tumors in the pathogenesis of autoimmune and the origin of the autoantibodies in paraneoplastic pemphigus (PNP) in recent years. A Castleman’s tumor from a patient was identified to produce autoantibody. To identify the relationship between the tumor and pathogenesis of the disease, we analyzed the rearrangement of immunoglobulin variable region gene and its hypermutation in B cells of Castleman’s tumor from a patient who was diagnosed of paraneoplastic pemphigus. Methods: The surface-markers of cultured tumor lymphocytes were assessed with immunochemistry staining. After total RNA of the tumor cells were isolated, the mRNA was reversely transcribed into cDNA. V H and V L genes were cloned and their sequences were analyzed. Results: Immunochemistry staining and flow cytometer analysis showed that the tumor cells were CD20, HLA-DR, smIgM, and smIgG positive. The cloned IgV H and IGHV3-9*01 germ-line gene are homologous and so are the Ig V L and the IGKV4-1*01 germ-line gene. More nucleotide changes in the V H or V L occurred in CDRs than those in FRs. Conclusion: In this reported case, a clone of specific B-lymphocyte in the Castleman’s tumor carrying functional rearranged immunoglobulin heavy and light chain genes was found to have experienced switch recombination and was possible to produce IgG autoantibody.

Objective To observe of different facemask pressure controlled ventilation yongon gastric insufflation evaluated by ultrasound in infants during anesthesia induction.Methods Sixty ASA Ⅰ infants aged 1-3 yr,undergoing elective surgery,were randomly assigned to three groups ac-cording to facemask ventilation pressure:10 cm H 2 O (P10),1 5 cm H 2 O (P1 5 )and 20 cm H 2 O (P20)with twenty in each group.Infants were injected with propofol 2 mg/kg,fentanyl 0.002 mg/kg,cis-atracurium 0.1 5 mg/kg for general anesthesia induction until consciousness lost,then face-mask pressure controlled ventilation was applied for 120 s.Some respiratory parameters (SpO 2 , PET CO 2 )were recorded at the time of loss of consciousness (T0 )and after facemask pressure con-trolled ventilation for 30 s(T1 ),60 s(T2 ),90 s(T3 ),120 s(T4 )and after tracheal intubation(T5 ). The cross-sectional transverse and longitudinal diameter and area were measured respectively using ul-trasound at T0 and T4 .Results In all groups,SpO 2 was greater than or equal to 99% at all time points.PET CO 2 at T1-T5 was significantly higher than that at T0 and PET CO 2 at T5 was higher than that at T4 in all three groups.There were statistically significant increases in the values of the antral cross sectional area before and after facemask pressure controlled ventilation in group P20 (P <0.05). Conclusion During anesthesia induction in infants,1 5 cm H 2 O facemask ventilation pressure can guarantee adequate ventilation,and avoid gastric insufflation.

BACKGROUND:Conventional methods,including trypsin digestion and cells transfer using single call suspension,have many drawbacks,which limit the development of bone tissue engineering.OBJECTIVE:To culture bone marrow stromal stem calls,induce osteoblastic differentiation,and prepare cell sheets.METHODS:Canine bone marrow stromal calls were isolated by density gradient centrifugation technique,inoculated into DMEM medium,and induced to differentiate into osteoblasts.Complete call sheets were harvested by call sheet engineering based on the temperature change of temperature-responsive medium.RESULTS AND CONCLUSION:Immediately after inoculation,primary calls were scattered on the bottom of culture flask,presenting a transparent spherical body with a good refractive capacity.At 12 hours,calls exhibited a long shuttle shape,reached complete confluency,and grew in a whirlpool-like fashion.After osteoblastic induction,the majority of bone marrow stromal stem calls appeared tetragonal,polygonal,and squamose.At 21-28 days,round or oval-shaped calcified nodules formed.When the bone marrow stromal stem calls in the temperature-responsive culture dishes were cooled below the critical temperature 32℃,cells were gradually detached from the bottom of culture flask and formed complete bone marrow stromal stem call sheets.These findings indicate that density gradient cantrifugation technique can be used to successfully isolate and culture canine bone marrow stromal stem cells to differentiate into osteoblasts and call sheet engineering enables to harvest complete bone marrow stromal stem call sheets.

Objective To explore the curative effect of ursodeoxycholi acid on retinopathy of prematurity (ROP) caused by high concentration of oxygen in newborn rats. Methods The model of ROP was established. Neonatal rats were divided into normal control group, ROP model group, low dose ursodeoxycholi acid treatment group (10mg/kg) and high dose ursodeoxycholi acid treatment group (40mg/kg). Rats were sacrificed at days 17. The new retinal vessels were observed and counted under lfuorescence microscope. Results The new retinal vessels in ROP rats were hyperplastic, twisted and unevenly distributed. There was signiifcant difference in the number of new retinal vessels among different groups (P=0.000). The number of new retinal vessels of rats in ROP group, low-dose group and high-dose group was signiifcantly more than that in control group (P=0.000). The number of new retinal vessels in low-dose group and high-dose group was significantly less than that in ROP group (P<0.05). The number of new retinal vessels in high-dose group was less than that in low-dose group (P>0.05). Conclusions Ursodeoxycholi acid could inhibit the angiogenesis of retina and could have curative effect on ROP.

Objective] To analyze the sequence difference of the SSU rDNA variable regions of Leishmania isolates from hilly foci and plain foci of China. [Methods] Specific SSU rDNA fragments from nuclear DNA of five Leishmania species and isolates were amplified by PCR. The amplified DNA fragments were cloned into pGEM R\|T Easy vector. The specific fragments were sequenced by the automated DNA sequencer. [Results] Sequence analysis showed that the amplified DNA fragments of five Leishmania species and isolates were all 392 bp in length, point mutations were located in the two unique sequence (UQ\|Ⅰ and UQ\|Ⅱ); L.d.SC10 and L.d.GS7 had two same point mutations in UQ\|Ⅱ, only L.d.GS7 had one in UQ\|Ⅰ; no insertion/deletion. [Conclusion] Sequence difference of the SSU rDNA variable region existed between Leishmania isolates from hilly foci and plain foci; The sequences of the SSU rDNA variable regions of L.d. SD2 isolate and L.infantum were identical.

Objective To study in vitro the inhibitory effects and mechanisms of N-butanol extract of Potentilla anserine L.(NP)against hypoxia-induced nitric oxide(NO)in hippocampus neuron of rats. Methods The models of hippocampus neurons hypoxia injury of Sprague-Dawley(SD)neonatal rats were cultured in vitro. The cultured hippocampus neurons were divided randomly into blank control group, hypoxia injury model group, nimodipine group(2 μmol/L)and NP high(250.0 mg/L),middle(62.5 mg/L),low(15.6 mg/L)dose groups. The activities of hippocampus neurons were examined by methyl thiazolyl tetrazolium(MTT)assay,and meanwhile their contents of nitrogen monoxidum(NO)were detected. Half quantity reverse transcription-polymerase chain reaction(RT-PCR)and Western blotting were used to detect neuronal nitric oxide synthetase(nNOS)mRNA and protein expression levels respectively in each group,immunocytochemistry stain was used to detect protein positive rate. Results Compared with blank control group,the activity of neuron〔absorbance(A)value〕was significantly decreased(0.0826±0.0095 vs. 0.3315±0.0105),content of NO(μmol/g:0.0509±0.0027 vs. 0.0291±0.0032), the expression levels of nNOS mRNA (0.1463±0.0081 vs. 0.0801±0.0058), the positive rate of nNOS〔(74.4238±3.9423)%vs.(28.3714±4.1361)%〕,the expression levels of nNOS protein(A value:1.9130±0.0471 vs. 0.5068±0.0368)were all significantly increased in the hypoxia injury model group(all P<0.01). Compared with hypoxia injury model,the activity of neuron was increased,contents of NO,the expression levels of nNOS mRNA,the positive rate of nNOS,the express levels of nNOS protein were decreased in each medicine group,especially prominent in the NP high concentration group〔the activity of neuron:0.1681±0.0118,contents of NO:0.0319±0.0044,nNOS mRNA:0.0648±0.0032,nNOS positive rate:(40.1240±6.4900)%,nNOS protein:1.3924±0.0621,all P<0.01〕. There were no statistical significant differences between the NP low concentration group and model group(all P>0.05). Conclusions NP can ameliorate the injury of rat hippocampus neurons induced by hypoxia in vitro. The possible mechanisms might be related to the effective inhibition of the synthesis of nNOS and NO excessive generation.

Objective To explore the impacts of enriched environment(EE),which has different initiation time points and intensity,on the neural and ethological prognosis and contents of myelin basic protein(MBP)of neo-natal rats with hypoxic - ischemic brain damage(HIBD). Methods HIBD rat models were established. Rats were divided into the early,the intermediate and the late intervention groups,which experienced EE from 7,14 and 21 days after HIBD for 14 days. The early and intermediate intervention groups were then divided into 6 - h and 24 - h groups, which experienced EE intervention for 6 hours or 24 hours respectively each day. Trapeze tests and water maze tests were carried out to detect the neural and ethological prognosis. Immunohistochemical method was used to detect MBP of the brain white matter,and the percentages of positive cells with MBP were detected by an image analyzer. The con-tents of MBP were measured. Results The trapeze test scores of the early and intermediate sham operation group,HI group,early 6 - h and 24 - h EE groups and the intermediate 6 - h and 24 - h EE groups,the late sham operation group,the late HI and late EE intervention group were(4. 05 ± 0. 88)scores,(2. 35 ± 1. 02)scores,(3. 67 ± 1. 12) scores,(3. 50 ± 1. 41)scores,(3. 50 ± 0. 93)scores,(3. 56 ± 1. 13)scores,(4. 00 ± 0. 89)scores,(2. 17 ± 1. 17)scores,(3. 50 ± 0. 92)scores,respectively. The trapeze test scores of early,intermediate and late EE groups were higher than those of the HI groups in the same period. There was no significant difference between the early,the intermediate 6 - h EE groups and 24 - h EE groups. Scores of water maze of each corresponding group were(40. 68 ± 23. 77)seconds,(56. 66 ± 10. 96)seconds,(46. 49 ± 19. 27)seconds,(51. 72 ± 20. 46)seconds,(38. 20 ± 18. 36)seconds,(47. 96 ± 20. 65)seconds,(38. 63 ± 20. 44)seconds,(59. 66 ± 13. 81)seconds and(45. 93 ± 22. 45)seconds,respectively. The water maze scores of the early,the intermediate 6 - h EE group and the late 24 -h EE groups were higher than those of the HI groups in the same period. There was no significant difference between the early,the intermediate 6 - h EE groups and the 24 - h EE groups. The relative abundance of MBP of the early and intermediate and the late HI groups were 6. 32 ± 1. 63 and 6. 74 ± 2. 19,and significantly less than that of the sham groups in the same periods,which were 9. 09 ± 1. 69 and 9. 37 ± 2. 46. The relative abundance of MBP of early 6 - h and 24 - h groups,the intermediate 6 - h and 24 - h groups and the late EE group was 7. 84 ± 2. 51,8. 05 ± 1. 86, 8. 89 ± 2. 29,8. 48 ± 2. 67 and 7. 98 ± 2. 09,respectively,which was significantly higher than that of the HI groups in the same periods. It showed that the neural and ethological prognosis of neonatal rats with HIBD could be improved,no matter the intervention began in the early,the intermediate or the late periods,or the intervention time was 6 hours or 24 hours each day. And relative abundance of MBP in the white matter increased with EE. Conclusions EE interven-tion has a long window stage for young rats. EE intervention could improve the neural and ethological prognosis of rats with HIBD. EE intervention could elevate the contents of MBP in the white matter,which could be one of the mecha-nisms for EE to improve the neural and ethological prognosis of rats with HIBD.