Female ; Humans ; Infant ; Metabolism, Inborn Errors ; Methylmalonic Acid ; blood

Objective To screen the proteins binding with HBV X-promoter and to investigate their potential role in the regulation of replication and expression of HBV DNA. Methods By using HBV X biotinylated promoter DNA as the selective molecule, the T7 select human liver cDNA library was biopanned and positive clones were selected. After screening, positive plaques were performed to amplify for inserted DNA fragment and cloned into pGEM-Teasy vector. Thirty positive plaques were chosen for DNA sequencing. Results Five kinds of known and nine kinds of novel cDNA sequences were obtained. The 5 kinds of known ones are human serum albumin, NADH dehydrogenase subunit 4, prokininase, ubiquitin specific protease 10, and testis enhanced gene transcript. Conclusion The binding proteins of HBV Xp DNA were identified by phage display. The results suggest that this approach provides a new search tool for the study of replication and expression mechanism of HBV DNA.

ABSTRACT:Objective To observe the anesthetic effects of xylazine and dexmedetomidine in establishing the model of spinal cord injury in beagle dogs.Methods Thirty female beagle dogs were randomly divided into 3 groups with 1 0 in each:xylazine group (group S ), dexmedetomidine group (group D ), and xylazine +dexmedetomidine group (group S+D).Basal anesthesia with ketamine was performed in all the dogs.Then group S was intramuscularly injected with xylazine discontinuously, group D was intravenously injected with dexmedetomidine 2μg/(kg·h)continuously,but group S+D was injected with xylazine and dexmedetomidine in combination.The heart rate (HR),respiratory frequency (RR),Ramsay score and spasm index (SI)were observed at the time of skin incision (T1),after the beginning of the operation 30 min (T2),before the end of the operation 30 min (T3),and the end of the operation(T4);and the usage of anesthetics was recorded.Results Compared with that in groups S and D,the usage of xylazine and dexmedetomidine in group S+D was decreased by 1/2 and 1/4,respectively (P<0.01),but HR and RR did not change significantly;the sedative effect was superior to that in S.No obvious increase in muscular tensility in the foreleg was observed before operation.Conclusion Xylazine and dexmedetomidine used in combination to establish the model of spinal cord injury in beagle dogs have advantages of better sedative effect,less respiratory depression and lower dosage of anesthetic drugs.

Objective To explore the fluctuation of cytokine mRNA expression level in a novel T-cell-mediated immune hepatic fibrosis model induced by repeatedly injections of Concanavalin A in BALB/c mice. Methods BALB/c mice were divided into different groups. Model group mice were injected weekly up to 20 weeks with Concanavalin A (15mg/kg), via retro-orbital venous plexus under ether anesthesia. Normal control group mice were treated in the same manner weekly with normal saline. Twenty-four hours after Concanavalin A challenge at 1, 5, 12 and 20 week, 8 mice from each time were killed by cervical dislocation, repectively. The livers of different group were excised and fixed in 10% formalin for HE staining and Gomori Ag staining or frozen in optimal cutting temperature (O.C.T.) media in liquid nitrogen for immunohistochemical staining for CD4 +T or CD8 +T cell. After extracting total RNA from liver tissues, IL-2, IL-4, IL-10 and transforming factor ?1 messenger RNA were amplified by reverse transcription polymerase chain reaction. PCR products were electrophoresed on agrose containing ethidium bromide and visualized under ultraviolet light. Densitometric RT-PCR data were standardized with ?-actin signals. Results The histological change of HE staining and Gomori Ag staining indicated the fibrogenesis in model group mice. Immunohistochemical staining for CD4 + or CD8 + T cell indicated that the infiltrating lymphocytes in liver parenchyma were mainly CD4 +T lymphocytes. IL-2 mRNA expression level only increased after the first injection of Concanavalin A. The expression levels of IL-4, IL-10 and transforming growth factor ?1 mRNA significantly increased over the whole experiment period as compared with control group. Conclusions Repeated administration of Concanavalin A can induce T-cell-mediated immune hepatic fibrosis model in BALB/c mice. The expression levels of IL-4, 10 and TGF-?1 increase over the whole experiment period and may play an important role in creating mouse fibrotic model.

Objective To give a summary of the current researches on hematopoietic stem cell niche.Methods Through extensive reviewing the related domestic and abroad literatures,the present summary reviews the compo- nents of hematopoietic stem cell niche,related cell factors and related cell signaling pathway participating in regula- tion of hematopoietic stem cell.Results The activities of hematopoietic stem cells are regulated by the niche(micro- environment),which is composed of hematopoietic stem cell and its surrounding cells.The regulation cannot be completed through one signaling pathway.Also,the self-renewal and differentiation of hematopoietic stem cell can- not be completed only through osteoblastic cells.Conclusion The niche regulates hematopoietic stem cells by differ- ent ways.With study in-depth,we will comprehensively understand the nature of stem cells and the study will pro- vide a broader space to stem cell-based therapy.

Objective To explore the clinical characteristics and diagnosis of mycoplasma pneumoniae(MP) pneumonia showing lobar pneumonia.Methods Patients from Jan.2004 to Dec.2005 were selected as researching case,which had samptoms and signs of respiratory in clinic,the chest Xray shown lobar pneumonia,the MP of down respiratory′s secretion detected by polymerase chain reaction(PCR) and MP-IgM of serum detected by indirect hemagglutination were positive.The clinical data of them were reviewed.Results MP pneumonia showing lobar pneumonia was same to streptococcus pneumoniae pneumonia in clinic,but it had some characters:1.majority of the lobar MP pneumonia were children being student;2.the process was long,the samptoms of respiratory was tipical,the continuous fever was few,and the infectious samptoms was not obvious as streptococcus pneumoniae pneumonia;3.the injury to lung was more and tipical;4.the increase of white blood cell,neutrocyte and C-reactive protein were not clearly as to streptococcus pneumoniae pneumonia,but the absorbing of lung′s focal was relatively fast;5.the ?-lactams antibiotic was not effective,but macrolides was effective.Conclusions Clinical characters of MP pneumonia are needed to master,the variety of chest X-ray changes and differentiate from other pneumonia infected by other are needed to notice.

Objective To understand the regional distribution and drug resistance of clinical isolates of bacteria flora ,in order to provide the basis for the diagnosis and treatment of bacterial infection .Methods According to the national clinical test proce‐dures operation separation strains ,HX‐21 bacteria identification/susceptibility analyzer bacteria identification and drug sensitive test .Results Among 1 705 strains of isolated bacteria ,Gram‐positive cocci accounted for 39 .8% ,Gram‐negative bacteria accounted for 60 .2% ;separation rate from high to low were:benzene azole resistance westwood 13 .3% coagulase negative staphylococcus , pseudomonas aeruginosa ,12 .0% benzene azole resistance westwood staphylococcus aureus 11 .3% ,did not produce the ultra broad spectrum beta lactamase 10 .7% ,e .coli to produce ultra broad spectrum beta‐lactamase e .coli 9 .9% ,acinetobacter was 7 .1% ,pro‐ducing ultra broad spectrum beta‐lactamase pneumonia klebsiella bacteria 5 .9% ,did not produce the ultra broad spectrum beta‐lac‐tamase pneumonia klebsiella bacteria 5 .7% ,benzene azole westwood sensitive coagulase negative staphylococcus 4 .8% ,enterococ‐cus was 4 .8% ,benzene azole westwood sensitive staphylococcus aureus 4 .5% ,etc .Conclusion The common pathogenic bacteria is gram‐negative bacilli in the common pathogenic bacteria .In negative bacilli infection ,acinetobacter ,pseudomonas aeruginosa ,e .coli , klebsiella pneumoniae ,etc were more common .In staphylococcus aureus strains ,benzene azole westwood drug‐resistant strain ratio is higher than benzene azole westwood sensitive strain rate for 3 times .In addition to the vancomycin and teicoplanin sensitive ,other commonly used antibiotics shows different degrees of resistance .

Objective To investigate the effect of retinoblastoma binding protein 4 (RBBP4)in Sp1 -mediated HIV long terminal repeat(LTR)transcription.Methods RBBP4 expression vector and Sp1 expression vector were respectively co-transfected into 293 T cells with HIV promoter pHIV-LTR-Luc or Sp1 site mutated pHIV-LTR-sp1 -mut by liposome transfection,and the transfected cells were examined by dual luciferase reporter assay system.The effect of RBBP4 on the binding of Sp1 to LTR was further studied by chromatin immunoprecipitation (ChIP)and electrophoretic mobility shift assay (EMSA).Results The relative firefly luciferase activity activated by Sp1 was decreased from 62.5 to 16 at the dose of 500 ng of RBBP4 expression vector (t =14.52,P <0.01 ).When the Sp1 binding sites were mutated,the effects of 100,300 or 500 ng of RBBP4 expression vector on the firefly luciferase activity of HIV LTR were not statistically significance (t =1 .897,2.357 and 3.162,all P <0.05).ChIP results showed that when the binding of RBBP4 on HIV LTR increased,the binding of Sp1 on HIV LTR increased significantly (t =11 .93,P <0.01 ),while the reduced binding of RBBP4 on HIV LTR significantly attenuated the binding of Sp1 onto LTR(t =11 .38,P <0.01 ).The effect of RBBP4 on the binding of Sp1 to DNA in ChIP assays was further verified by EMSA assays.Conclusion RBBP4 can inhibit the Sp1 -mediated HIV LTR transcription in 293 T cells.

for the detection of their virulent abilities simultaneously.

Objective To detect the mechanism of serum uric acid (UA) in different types of coronary heart disease (CHD).Methods 88 patients were divided into three groups：the control group，stable angina (SA) group and acute coronary syndrom (ACS) group.The levels of UA，alpha-granule membrane protein 140 (GMP-140)，von Willebrand factor(vWF)，plasminogen activator inhabitor typed (PAI-1)，Thromboxane B_2 (TXB_2) and C-reacting protein(CRP) were measured.Results ①UA and CRP in ACS group were higher than that in SA group and control group [(392.1±68.57) μmol/L and (42.2±39.4) mg/L vs (370.50±58.80) μmol/L and (18.9±17.1) mg/L vs (286.00±65.31) μmol/L and (2.5±0.7) mg/L，P＜0.05)].For UA，there was no difference between ACS and SA group(P＞0.05)；CRP was higher in ACS group than in SA group (P＜0.05).②vWF and TXB_2 were higher in ACS[(1.65±0.48)%，(19.73±18.66)ng/L]and SA group[(1.35±0.49)%，(11.18±10.71) ng/L]than in control group[(1.07±0.26)%，(6.46±5.41) ng/L，P＜0.05]，and those were higher in ACS group than in SA group (P＜0.05).③GMP-140 and PAI-1 were higher in ACS [(13.04±0.99) μg/L and (65.65±14.76) μg/L]and SA group[(12.55±0.74) μg/L and (62.69±12.24) μg/L]than in control group [(12.32±0.29) μg/L，(50.78±13.88) μg/L，P＜0.05].There were no differences between ACS and SA group (P＞0.05).④Comparing hyperuricemia group and non-hyperuricemia group in CHD patients：the CRP(71.3±18.9) mg/L，vWF(1.08±0.52) %，GMP-140(13.57±1.11) μg/L，TXB_2 (57.26±47.84)ng/L，PAI-1 (72.12±9.23) μg/L in ACS group possessing hyperuricemia were higher than non-hyperuricemia group [CRP (20.7±17.9) mg/L，vWF (0.84±0.54) %，GMP-140 (13.23±1.07) μg/L，TXB_2 (26.70 + 23.83) ng/L，PAI-1 (61.30±12.07) μg/L](t=7.394，0.008，0.227，7.605，0.421，P＜0.05)；CRP(31.1±18.9)mg/L and TXB2 (21.54±3.90) ng/L in SA group possessing hyperuricemia group were higher than non-hyperuricemia group[(10.9±10.1)mg/L and (5.02±4.93) ng/L，t=0.494，8.669，P＜0.05].Logistic stepwise regression analysis indicated that the related factors with ACS were UA(OR=1.046)，CRP(OR=7.615)，PAI-1(OR=1.301)，PT(OR=0.300)and TG(OR=2.243) (P＜0.05).Conclusions UA is an important risk factor in CHD patients.UA can induce different types of CHD by damaging blood vessel endothelium function，activating platelet，changing coagulation and causing inflammatory.