Post-translational modification by ubiquitin and ubiquitin-like modifiers(Ubls) is one of the most important mechanisms regulating a wide range of cellular processes in eukaryotes.Previous research showed that,through covalently modification by ubiquitin or ubls,the substrate proteins can be regulated in many different ways like stability,subcellular localization,enzymatic activity,protein-protein interaction and so on.Therefore,we believe,that ubiquitin and ubls play very important roles in cellular and biological processes by modifying plenty of proteins.To better understand the ubiquitin and ubls system,proteomic approaches have been developed to purify and identify more protein substrates.Large-scale idendification of ubiquitin/ubls-modification sites by mass spectrometry is particularly important for understanding the molecular mechanism and function of ubiquitin/ubls modification.Upto the present,more and more scientists are getting interested and participating in proteomics research of ubiquitin/Ubl modifications.This review summarizes the rencent results in this field.

Objective To detect the expression of Zbtb7 gene in Phorbol esters(PMA) induced differentiation of leukemia cell lines.Methods 50 nmol/L PMA was used to induce the differentiation of U937 and K562 cell lines.The expression of Zbtb7 was detected by real-time PCR in the cells after induction on day 0,1,3 and 5,respectively. Results The expression of Zbtb7 in leukemia cell lines was markedly enhanced after treated by PMA(P

Objective To analyze the characteristics of blood loss for laboratory test of critically ill premature infants,and to seek feasible measures to reduce the blood loss.Methods Two hundred and forty-six cases of critically ill premature infants admitted to the neonatal intensive care unit from April 2012 to April 2013 were analyzed the blood loss for test during the hospitalization and the clinical features of blood loss with different gestational age,different weight within the first four weeks after admission.Then the application of blood loss according to test category was described.The blood volume demanded in theory was determined by the formula B =5 (∑ s + 0.1),then calculated the phlebotomy overdraw on the basis of the practical blood loss and analyzed the characteristics of overdraw per patients per day in first two weeks after admission.Results Among 246 patients,The median blood loss figure was 25.57 ml for each infants with the range between 7.10 ml ～ 119.20 ml,and the blood loss concentrated in first four weeks,which showed a decreasing trend with time.There was a statistically significant difference(P ＜0.05)that the smaller gestational age,the lower birth weight,the more daily blood loss per patient per day in first two weeks,but no significant differences(P ＞ 0.05) between the third and fourth week.The largest proportion of the blood samples was used for clinical chemical tests(31.49％),followed by blood gas analysis (19.03％),immunoassays (12.69％),blood cultures (12.63％),hematology (12.28％).The practical blood loss was about twice times of blood volume demanded for tests in theory,which the median was 7.8 times to the latter(25.57 ml vs 3.26 ml).It showed statistically significant difference(P ＜0.05) between blood overdraw per patients per day in the first week and the second week.Compared with different gestational age and birth weight,the difference of overdraw was also statistically significant(P ＜0.05).According to test category,blood culture was the most significant samples of phlebotomy overdraw,followed by biochemical,other,blood gas analysis,the percentage was 76％,64％,45％ and 41％ respectively.Conclusion The blood loss for laboratory test and the phenomenon of blood waste is serious in critically ill preterm infants.The smaller the gestational age is,the lower the weight is,the amount of blood loss and phlebotomy overdraw are more significantly.Biochemical and blood gas analysis are the main items of blood loss.

Dihydroartemisinin (DHA),the major active metabolite of artemisinin,participates in tumor progression through the following ways:forming free radicals to induce cancer cells death dependent on iron,inducing apoptosis,inhibiting angiogenesis,tumor cells invasion and metastasis,modulating muhidrug resistance,controlling intracellular Ca2+ concentration,regulating cell cycles,cell autophagy and the immune system and so on.Generally,it is considered to be a potential anti-tumor drug.

Recombinant protein SMB(PRG4) containing two Somatomedin B domains and a small amount of glycosylation of repetitive sequences of proteoglycan 4 was cloned according to PGR4 gene polymorphism. Mature purification process was established and recombinant protein SMB(PRG4), with high-level expression was purified. By using size-exclusion chromatogaraphy and dynamic light scattering, we found that the recombinant protein self-aggregate to dimeric form. Structure prediction and non-reducing electrophoresis revealed that SMB(PRG4), was a non-covalently bonded dimer.
Glycosylation ; Protein Multimerization ; Proteoglycans ; chemistry ; Recombinant Proteins ; chemistry ; Somatomedins ; chemistry

BACKGROUND: Hippocampal neuron presents remarkably injury in cerebral after seizure of epilepsy. Necrosis and apoptosis are two kinds of neural cell injury after epilepsy and play an important role in neural injury of epilepsy. Being endogenous neural protective transmitter, adenosine may inhibit the release of excitatory amino acid, production of oxygenic free radical and action of nitric oxide. Simultaneously, it can improve cerebral blood flow and anti-convulsion. But it has been unknown concerning to the relationship between adenosine and cell apoptosis after epilepsy yet.OBJECTIVE: To observe the effects of 2-CAdo adenosine-receptor excitant on genetic expression of bcl-2, Bax of hippocampal cells in epileptic rats and further probe into the mechanism of adenosine on anti-convulsion and brain protection.DESIGN: Completely randomized controlled experimental research in which the experimental animals were taken as the objects.SETTING: Pediatrics department and general surgical department of one oil field general hospital, and pediatric internal department of a hospital affiliated to one university.MATERIALS: The experiment was performed in Experimental Zoology Departnent and Pathological Teaching & Research Department of Harbin Medical University from October 2002 to March 2003. Totally 104 Wistar rats of either sex were employed, weighing varied from 200 g to 250 g. The animals were randomly divided, named as normal group 8 rats, epileptic group 32 rats, epileptic & 2-CAdo group 32 rats, and epileptic & physiological saline group 32 rats.INTERVENTIONS: The animal epileptic model was set up by intra-abdominal injection of coriamyrtin 15 mg/kg(provided by Pathology Department of Harbin Medical University. Convulsion presented in all of rats, 5 minutes later after injection, lasting for 1 or 2 minutes. In epileptic & 2-CAdo group, 2-CAdo(provided by ICN company), 0.6 mg/kg, was injected from the vein on the tail 1 hour before coriamyrtin injection and 1 hour after convulsion respectively. In epileptic & physiological saline group, the physiological saline of equal dosage was injected from the vein on the tail 1 hour before coriamyrtin injection and 1 hour after convulsion respectively.MAIN OUTCOME MEASURES: Positive cell counts of bcl-2 and Bax genetic expression in hippocampal CA1 area.RESULTS: Twenty-four hours after epilepsy seizure, neural cell bcl-2 expression was increased in hippocampal CA1 area, was remarkably decreased in 48 hours, and the expression was only little amount in 72 hours, but it was increased again in 7 days. Bax expression began increased in 24 hours after epilepsy seizure, was significantly increased in 48 hours, reached the peak in 72 hours, the expression was the minimum in 7 days. In epileptic & 2-CAdo group, bcl-2 expressions at corresponding times were remarkably increased compared with epileptic group and epileptic & physiological saline group( P＜ 0.05), Bax expressions were remarkably decreased compared with epileptic group and epileptic & physiological saline group( P ＜ 0.05), indicating statistical significance.CONCLUSION: 2-CAdo can reduce apoptosis of hippoeampal neural cells after epilepsy seizure and provide a certain protection for neural cells.

Objective To establish a reliable unstable pelvic fracture model with sacral plexus stretch injury in goats and to detect electrophysiological signals when pelvis is shifted in different orientations and distance.Methods Twenty-four goats were divided into three groups of 8 animals each:pelvic longitudinal displacement was induced in group A,pelvic lateral displacement in group B,and pelvic dorsal displacement in group C.Animal left L6 and S1 nerve root and initial segment of sciatic nerve were exposed to connect stimulating and receiving electrodes respectively.Left pelvis was shifted at the speed of 0.25 mm/s to produce the model of unstable pelvic fracture with sacral plexus stretch injury.Electrophysiological signals of L6 and S1 were recorded when the placement of pelvis was 0,0.5 and 1.0 cm respectively.Results With pelvic displacement,the threshold potential,maximum stimulus potential and incubation period were gradually increased,but the peak-to-peak value became smaller gradually.When the pelvis displaced 0.5 cm at different orientations,group A showed improved threshold potential and incubation period (P ＜0.05),decreased peak-to-peak value (P ＜0.05),and no significant change in maximum stimulus potential(P ＞0.05);group B only showed lowered peak-to-peak value with no significant variations in other electrophysiological signals (P ＜ 0.05);group C had no significant variations in all electrophysiological signals (P ＞ 0.05).When the pelvis displaced 1.0 cm at different orientations,groups A and B showed improved threshold potential,maximum stimulus potential and incubation period (P ＜ 0.05) and lowered peak to peak value (P ＜ 0.05);group C showed higher incubation period with no significant variations in other electrophysiological signals(P ＜0.05).Between groups,the amplitude of variation in electrophysiological signals was the most in group A and the least in group C.Conclusion After unstable pelvis fracture in goats,sacral plexus stretch injury is the most serious in longitudinal displacement of the pelvis.

Objective The aim of the study was to detect the mutation of Fms-like tyrosine kinase 3 internal tandem duplication (FLT3-1TD) rate in older de novo acute myeloid leukemia (AML) patients, to evaluate the role of FLT3-ITD in AML and its clinical significance. Methods The mutations of FLT3-1TD in bone marrow mononuelear cells (MNCS) from 30 cases of older AML were screened by polymerase chain reaction denaturing-high performance liquid chromatography (PCR-DHPLC). Results FLT3-1TD mutations were identified in 26.67 %(8/30) patients, while there were no mutations identified in control cases. And these kinds of mutations were likely to attend in M3 types. All mutations of FLT3-ITD were heterozygous and rearrangement fragment located in reading frame. Different karyomite groups had different FLT3-ITDmutations rate. We could see that FLT3-ITD positive patients were more prevalent in patients with normal karyotype. Clinical researches indicated that FLT3-ITD mutations had the characteristics of a higher peripheral white cell count, higher blast cells and lower complete remission rate in older AMKA Conclusion FLT3-ITD positive older AML patients conferred a poor prognosis and were likely to attend in normal karyomite group. The detection of FLT3-ITD mutations could make up for the deficiency of cytogenetics to some extent, and may become a routine examination of AML in older, which can direct their treatment and predict their prognosis.

Objective To express the human recombinant SUMO1 protein and prepare monoclonal antibody(mAb) against it.Methods The recombinant expression plasmid pET32a-HIS-SUMO1 was made and transformed into E.coli(BL21),then the recombinant fusion protein HIS-SUMO1 was expressed and purified.The BALB/c mice were immuned with pure protein HIS-SUMO1 as antigen.Monoclonal antibody against SUMO1 was prepared with standard hybridoma technology.The hybridoma cell lines were obtained by ELISA and Western blot screening procedure,the isotype of the mAbs were further identified by immune-double diffusion.Ascites were collected from one propagated hybridoma cell line and mAbs were purified by using the Kit of Millipore.The valence of mAb was detected by Western Blot.Results The recombinant protein HIS-SUMO1 is expressed and purified.Three hybfidmas producing antibodies against SUMO1 were obtained,the isotypes of three mAbs are IgG1,Western blot showed that the antibodies were specific for SUMO1.The antibody purified from the ascites has better specificity.Conclusion The SUMO1 mAb prepared by using recombinant SUMO1 protein as antigen can be used for detectingthe protein sumoylation.

Ob jective To construct and screen out the RPL 23-siRNA interference fragments ,providing the basis for the following experiments about the correlation with RPL 23 and gastric cancer .Methods The RPL23-siRNA,synthesized chemically through lipofection ,were selected from three target sequences by RNA in-terference and detected by real -time PCR and Western blot .Results Compared with normal cell group and RPL23 control group ,the mRNA and protein expression of RPL 23 in the other 3 interference groups were signifi-cantly decreased(P<0.01).Multiple comparisons showed that the interference efficiency of RPL 23 -siRNA1 group was significantly higher than that of RPL 23-siRNA2 group and RPL23-siRNA3 group(P<0.01).Con-clusion The RPL23-siRNA interference fragment can be successfully constructed and screened out ,which pro-vides the basis for the following experiments .