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: 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.

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 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.

Background Genetic mutation remains to be the most common cause of congenital cataract.Whole exon sequencing technology is an ideal method to detect the pathogenic gene mutations.Objective This study was to identify the pathogenic gene in a Chinese autosomal dominant congenital cataract (ADCC) family by whole-exome sequencing.Methods This study complied with Helsinki Declaration and the protocol was approved by Ethic Committee of Peking University Third Hospital.Informed consent was obtained from each subject before any medical examination.A cross-sectional study was designed.A Chinese ADCC family with 4 generations and 48 members were enrolled in Peking University Third Hospital,of which Ⅰ1 and Ⅰ2 died.The periphery blood of 8-10 ml was collected from each member of Ⅱ,Ⅲ and Ⅳ generations for the high throughput sequencing of genes using whole exon trapping and new sequencing technology,and the sequencing results were compared with the data of human HA PMAP8,dbSNP130 and 1000 Genome Project database.The synonymous mutation was filtered after reported common variants,and the false positive results of explicit sequencing were finally excluded by Sanger sequencing and then the candidate genes were identified.The mutation genes were screened to determine the pathogenic gene of this ADCC family.Results Eleven ADCC patients were found in this family,and the patients distributed in each generation with an equal chance for involvement in male and female subjects,which conformed to an autosomal dominant inheritance pattern.All the patients were nuclear cataract.Genome-wide whole-exome sequencing found that major intrinsic protein (MIP) gene was known genes of ADCC in initially identified candidate genes,so the Sanger was used to verify the MIP gene.The heterozygous mutation of MIP gene (chr12:56845250 C ＞ T) appeared to be the pathogenic cause of this ADCC family.The mutation occurred in the splice sites of the gene,resulting in the fourth exon coded-61 amino acids are replaced by leucine,histidine and serine,which lead to the abnormal truncated proteins.Conclusions The heterozygous mutation of MIP gene is the molecular pathogenesis of this Chinese ADCC family.

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 analyze the risk factors associated with allergic disease in preterm infants during the first year of life. To investigate whether there was any association between peripheral blood eosinophils and allergic di-sease during the first year of life benefit of hydrolyzed formula feeding in preterm infants. Methods A nested case -control study was conducted in a cohort of artificial feeding preterm infants admitted to Neonatal Intensive Care Unit, Shanghai Children's Medical Center from January 2012 to April 2013. The preterm infants were monitored from birth up to 1 year of age and the findings were related to development of allergic disease. Individuals with allergic disease during the first year of life(cases)were matched with gender,as previously described with two individuals(controls)who re-mained event - free during the study. The risk factors associated with allergic disease in preterm infants during the first year of life were analyzed. And whether there was any association between peripheral blood eosinophils and allergic di-sease during the first year of life benefit of a hydrolyzed formula feeding in preterm infants was investigated. Different variables in control and case individuals were compared with t test for normal distribution measurement date,χ2 tests for categorical variables and Mann - Whitney U test for non - normal distribution measurement date,and multiple conditio-nal Logistic regression were used to investigate the risk factors associated with allergic disease in preterm infants. Results Thirty - four individuals were in cases and 68 individuals were in controls. In a conditional multivariable Logistic model,peripheral blood eosinophils of preterm infants at full enteral feeding(EOS - 2)(OR = 5. 941,95% CI:1. 165 - 41. 375,P ﹤ 0. 05),and family history of allergy(OR = 3. 316,95% CI:1. 201 - 9. 152,P ﹤ 0. 05)were the two independent risk factors for allergic disease in preterm infants during the first year of life. Individuals fed with standard preterm formula after birth,the association between EOS - 2 and allergic disease was significantly enhanced (OR = 21. 459,95% CI:1. 686 - 273. 152,P ﹤ 0. 05). By contrast,in individuals fed with hydrolyzed formula,the risk of EOS - 2 was substantially attenuated(OR = 1. 708,95% CI:0. 148 - 19. 743,P ﹥ 0. 05). Conclusions Peripheral blood eosinophils of preterm infants at full enteral feeding EOS - 2 and family history of allergy were the 2 independent risk factors for allergic disease in preterm infants during the first year of life. Contrast to individuals fed with standard preterm formula after birth,individuals fed with hydrolyzed formula had lower association between factors of peripheral blood eosinophils and family history of allergy,and allergic disease during the first year of life.

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.