AIM: To examine expression of macrophage migration inhibitroy factor (MIF) gene and protein in macrophages induced by oxidized low density lipoprotein (ox-LDL). METHODS: Macrophages were incubated with ox-LDL at the concentration of 150 mg/L for time course (0-36 h) and with ox-LDL at the different concentrations (0-300 mg/L) for 24 h, expression of MIF mRNA and protein were detected by RT-PCR and ELISA. RESULTS: The results showed that ox-LDL increased MIF gene and protein expression in macrophages in a dose and time-dependent manner. After the exposure of macrophage to ox-LDL, the expression of MIF mRNA level increased consistently with protein. CONCLUSION: MIF may play an important role in atherosclerosis. [

Objective To observe the effects and mechanism of pretreatment in rats with prostaglandin E1 on liver after hemorrhagic shock and resuscitation(HSR).Method In total,32 male SD rats were randomly(random number)divided into four groups(n=8):group A(sham group),group B(shock group),group C(HSR group)and group D(Lipo-PGEl+HSR).In group B,rats were sacrificed 90 min after shock,and in group C,rats were anesthetized and then subjected to hemorrhagic shock followed by resuscitation.In group D,rats were pretreated with Lipo-PGEI one hour before HSR.Liver function,NO and ET.1 were measured,and pathological changes of liver tissue in each group were observed,and the expres8ions of iNOS and ET.1 of liver tissue were measured by using immunohistochemistry 6 hours after HSR.Data were analyzed by analysis of variance,and P<0.05 was considered as significantly different in statitistics.Results The levels of liver iNOS and ET-I increased in HSR group compared with shock group [(O.225±0.080)vs.(0.082±0.021)and(0.292±0.047)vs.(0.082±0.035),P<0.05].Pretreatment with Lipo-PGEl markedly reduced the damage of Liver function,and lowered the levels of NO and ET-I.which were consistent with decrease in iNOS and ET-16 hours after HSR[(0.116±0.034)vs.(0.225±0.080)and(0.198±0.041)vs.(0.292±0.047),P<0.05].Conclusions Pretreatment with Lipo-PGEl could reduce liver injury after HSR.The mechanisms might be attributed to inhibiting iNOS and ET-1,regulating the balance of NO/ET-I.

tralateral parietal lobe. Conclusions CIMT can improve the patients' upper-limb function effectively. The constraint-induced movement of the affected arm during CIMT appears to induce cortical reorganization and compensation as measured by fMRL

Objective To construct and identify recombinant expression plasmid of small interfering RNA (siRNA)targeting hepatitis B virus X protein(HBx), and observe its effect on mitoehondrial function in healthy liver cell line steadily expressed HBx gene (HL-7702/HBx). Methods Two siRNA sequences containing short hairpin structure, which target on the total length HBx gene, were synthesized and cloned into the vector psiRNA-Hh1GFPzeo to eonstruct recombinant expression plasmids pX1 and pX2. Non-specific recombinant pScr plasmid served as control. After siRNA transfected into HL-7702/HBx cells line by liposome, reverse transcription-polymerase chain reaction (RT-PCR) and Western blot were performed to identify the suppressive effect on HBx expression. Levels of intraeellular reactive oxygen species (ROS) and mitochondrial membrane potential (△ m) were determined by flow cytometry. The experimental results were compared by analysis of variance. Results Successful constructions of pX1 and pX2 were confirmed by restriction enzyme digestion and sequencing. The expressions of HBx mRNA and protein after 48 h of transfection into HL-7702/HBx cells in control group were 0.65± 0.12 and 0.62± 0.09, respectively, which were both higher than those (0.33±0.10 and 0.19±0.08, respectively) in group pX1 (t=4.73, P<0.05; t=7.53, P<0.05) and those (0.48±0.10 and 0.37±0.11, respectively) in group pX2 (t=2.39, P<0.05;t=4.43,P<0.05). But the inhibition of group pX1 was stronger than that of pX2 (t=2.28,P<0.05). Levels of ROS and △ m after RNA interference were 5.00±0.38 and 33.86±0.50, respectively, while those in control group were 72. 10±0. 55 and 3. 57±0.26, respectively (ROS: t=276.22, P<0.05; △ m: t=107.15, P<0.05). Conclusions siRNA targeting HBx can efficiently and specifically suppress the HBx expression in HL-7702/HBx cells, and decrease the level of ROS and increase the level of △ m, thus relieve cellular oxidative stress.

Objective To observe the brain abnormalities of the white matter and myelination in patients with BH4 deficiency using MRI examination, and to further estimate the effect of treatment. Methods Eleven patients with BH4 deficiency aged 17 weeks to 4 years were observed, including 9 cases who were detected by newborn screening program. Although those patients were treated with low phenylalanine diet, and the blood phenylalanine 1evels maintained to 120-240 ?mol/L, the patients presented progressive hypotonia, convulsion, and mental retardation. All cases were diagnosed as BH4 deficiency by analysis of urinary pterins profile, BH4 loading test, and determination of dihydropteridine reductase in RBC. The patients received MRI examination using a 0.5 T MR system. Results Delayed myelination was found in 11 cases (100%) in frontal lobe, 8 cases (72.2%) in occipital lobe, 4 cases (36.3%) in temporal lobe, and 3 cases (27.3%) in parietal lobe. Delayed myelination of corpus callosum could be found in 6 cases (54.5%). There were abnormal diffuse high signals in the white matter on T_2WI in all cases. Conclusion Patients with BH4 deficiency demonstrate a high occurrence of brain abnormalities in the white matter. Those abnormalities are related not only with hyperphenylalaninemia, but also due to the decreased synthesis of neurotransmitters, such as L-Dopa and 5-HTP, all of these could be the reasons of adverse effect of the development of white matter.

Objective To assess the efficacy of mycophenolate mofetil (MMF) combined with low dose of hormone and lamivudine in the treatment of hepatitis B virus associated glomerulonephritis (HBV-GN).Methods The clinical data of 49 HBV-GN patients diagnosed by renal pathology was reviewed.They were treated with MMF( ≥ 12 years old,0.75 g,two times a day; ＜ 12 years old,0.4 g/m2,two times a day),low dose of hormone [0.5 mg/(kg·d) ],lamivudine( ≥ 12 years old,100 mg/d; ＜ 12 years old,3 mg/kg,two times a day).Results Among of 49 HBV-GN patients,clinical cure rate was 71.4％(35/49),the total effective rate was 81.6％(40/49),85.7％(42/49) patients' HBV DNA level decreased from 5.43 ×104 copies/ml to ＜ 1000 copies/ml.The complete remission rate was 88.0％ (22/25),the partial remission rate was 8.0％ (2/25),the inefficiency was 4.0％ (1/25) in membranous nephropathy (MN);the complete remission rate was 44.4％ (4/9),the inefficiency was 55.6％ (5/9) in mesangial proliferative glomerulone phritis (MsPGN) ; the complete remission rate was 70.0％(7/10); the partial remission rate was 30.0％(3/10)in membrane proliferative glomerulone phritis (MPGN) ;the complete remission rate was 40.0％ (2/5),the inefficiency was 60.0％(3/5) in focal segmental glomerulosclerosts (FSGS).There was significant difference among the different pathological type (P＜0.05).There were less adverse reactions.Conclusions MMF combined with low dose hormone and lamivudine is safe and effective in treating HBV-GN.The efficacy and pathological type has some relationship,MN has the best response,MPGN and MsPGN are second,FSGS is poor.

Objective To study the browning-related genes of Poria cocos mycelia,so as to provide a basis for their further verification and to discuss the browning molecular mechanism of Poria cocos.Methods Four samples of normal and brown Poria cocos mycelia were analyzed by transcriptome sequencing.After aligned with the reference genome sequence of Poria cocos,the expression levels of the genes in different samples were analyzed.Results A total of 12 383 transcripts were identified.Among them,1 017 genes were firstly found,and 260 genes were functionally annotated.Based on the alignment results,336 common differentially-expressed genes overlapped between the normal mycelia and 3 browned mycelia were obtained,and part of them were evaluated.Conclusion Transcriptome sequencing results showed that plenty of differentially-expressed genes were found in brown Poria cocos mycelia,and the difference of the expression levels of some genes was up to several hundred folds or even thousands of folds,whose related-functions were worth of further analysis.

BACKGROUND:Studies have shown that 26RFa plays an important regulatory role in bone formation, pain, endocrine, cardiovascular disease and energy metabolism. OBJECTIVE:To observe the effects of 26RFa on the proliferation and differentiation of human bone marrow mesenchymal stem cells. METHODS:In order to obtain the most efficient concentration of 26RFa, human bone marrow mesenchymal stem cells were detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide analysis. cells were inoculated into 6-wel plates and then divided into two groups:experimental group treated with 10-11 mol/L 26RFa and control group with no 26RFa. After 8, 12 and 16 days of osteogenic induction, alkaline phosphatase activities in induced cells were detected using alkaline phosphatase kit. After 21 and 28 days of osteogenic induction, alizarin red staining and Von Kossa staining were performed. The number of calcified nodules over each coverslip was calculated, and the expression of cbfa1 protein was detected by western blot assay. RESULTS AND CONCLUSION:After 8, 12, and 16 days of osteogenic induction, the alkaline phosphatase activities were higher in the experimental group than the control group (P<0.05, P<0.01, P<0.05). After 21 and 28 days of osteogenic induction, alizarin red staining and Von Kossa staining showed that the number of calcified nodules was higher in the experimental group than the control group, and the expression of cbfa1 protein was also higher in the experimental group (P<0.05). These findings indicate that 26RFa can promote the osteogenic differentiation of human bone marrow mesenchymal stem cells under appropriate culture conditions.

Objective To investigate the evidence of cortical reorganization in stroke hemiplegia treated with constraint-induced movement therapy(CIMT)by fMRI.Methods Five patients with chronic stroke were evaluated with the Action Research Arm Test(ARAT).The functional MRI(fMRI)was performed on a 3.0-T MRI with echo-planar imaging.The subjects were required to finish the finger-tapping task and undergo fMRI before and after CIMT.A block design was used for the inspection.Results After CIMT,the function of upper limb(sick side)of patients improved significantly assessed by ARAT(P<0.001),and cortical reorganization was found on fMRI.Conclusion CIMT can improve motor function of upper limb of chronic stroke patients with hemiplegia and induce cortical reorganization as measured by fMRI.

Objective To explore the interaction between hepatitis B virus(HBV) X protein and cytochrome C oxidase Ⅲ (COXⅢ).Methods Mating experiment and in vitro binding assays were utilized tO verify the interaction between HBV X protein and COX Ⅲ in vivo and in vitro respectively in yeast cells.Coimmunoprecipitation was employed to demonstrated the binding between these tWOproteins in mammalian cells.Results In mating experiment,the resulting diploids containing both pAS2-1-X and pACT2-COX Ⅲ plasmids turned blue in the β-gal assay,the result of Western blot displayed a positive reactive strap which indicated X protein could interact directly with COX Ⅲ in vitro binding assays.Interaction between these two proteins in mammalian cells was further confirmed by coimmunoprecipitation,as a protein strap about 17 000 was detected in Western blot.Conclusions It is speculated the specific interaction may exist between HBV X protein and COX Ⅲ in both prokaryotie and mammalian cells.X protein may impair the mitochondrial respiration chain and energy metabolism through its association with COX Ⅲ.