Objective:To discuss effect of nicorandil on unstable angina patients With persistent Weakly positive for troponin I (TnI).Methods:A total of 111 unstable angina patients With persistent Weakly positive for TnI Were randomly divided into control group (received routine treatment,55 cases) and intervention group (received nic-orandil 5mg,3 times/d based on routine treatment,56 cases).The relief of chest pain in one Week,the recurrent hospitalization for chest pain aggravation in 3 months and the cardiac mortality rate in one year betWeen tWo groups Were observed in tWo groups. Results:Compared With control group,the relief of angina pectoris in one Week (63.6% vs. 91.1%,χ2=11.97,P=0.0005)significantly increased,re-hospitalization for chest pain aggravation in three months (56.4% vs.19.6%,χ2=15.91,P=0.0001)significantly decreased in intervention group;but cardiac mortality rate during one year betWeen tWo groups Was no significant difference (5.5% vs. 8.9%,χ2=0.50,P=0.4792).Conclusion:Nicorandil can significantly reduce the unstable angina and re-hospitalization for chest pain aggravation in patients With persistent Weakly positive for TnI,but there Was no significant difference in reducing mortality Within one year betWeen tWo groups.

ObjectiveTo observe the effect of pulsed electromagnetic fields (PEMFs) on bone mineral density (BMD) of spinal cord injuried (SCI) patients. MethodsThe PEMFs group was composed of 12 SCI patients received regular PEMFs treatment as the treatment group, 12 SCI patients treated without PEMFs were as the control group.BMD of total proximal femur, Ward's trigonum and great trochanter of all patients were detected by dual-energy X-ray absorptiometry before and after treatment.ResultsBMDs of total proximal femur and great trochanter of patients of the treatment group were a little increased after PEMFs treatment, with no significant difference (P>0.05), but BMD change was significantly different compared with the control group (P<0.05～0.01).ConclusionPEMFs treatment along with routine exercise may help to reduce the loss of bone after SCI.

This study was carried out to predict the possible tertiary structure alterations of p53 protein after point mutation of p53 gene condon 282 in lung cancer cells based on the latest 3D structure analysis platform series of Phyre software. It was found that the p53 gene condon 282 mutation (Arg/Leu) may destabilize the H2 helix and DNA binding in the major groove by compromising the contacts of p53 protein with the beta-hairpin of DNA binding surface.
Amino Acid Sequence ; Base Sequence ; Codon ; genetics ; Humans ; Lung Neoplasms ; genetics ; pathology ; Models, Molecular ; Molecular Sequence Data ; Point Mutation ; Protein Structure, Tertiary ; Tumor Suppressor Protein p53 ; chemistry ; genetics

Objective To identify the role of phosphatidylinositol-3-kinase(PI3K) in mediating necroptosis induced by tumor necrosis factor alpha (TNFα) and the involved mechanism.Methods Knockdown of p110α,receptor-interacting protein 1(RIP1) or both p110αand RIP1 was mediated by the specific short hairpin RNA (shRNA) lentivirus and verified by RT-PCR or Western blotting .In addition , Western blotting was used to detect phosphorylation of mixed lineage kinase domain-like protein(MLKL) and protein kinase B(AKT) or tetramerization of MLKL.Cell death was measured by micros-copy and flow cytometry.Results AKT phosphorylation and TNFα-induced necroptosis of L929 cells were suppressed by the inhibitors of PI3K or AKT, as well as p110αknockdown.Moreover, RIP1 knockdown did not inhibit L929 cell death induced by TNFαplus Z-VAD, but the RIP1-independent necroptosis was inhibited by p 110αknockdown.In addition, p110αknockdown suppressed MLKL phosphorylation and tetramerization induced by TNFαwith Z-VAD in L929 cells. Conclusion PI3K mediates necroptosis of L929 cells induced by TNFαby activating AKT and MLKL, respectively.

Cathepsin B from Helicoverpa armigera (HCB) belongs to the group of cysteine proteinases. HCB is proved being involved in the degradation of yolk proteins during embryonic development,which is an acidic preferring enzyme and is resistant to SDS. The expression of the proenzyme may offer a model for investigating the activation of the enzyme. The HCB gene was constructed into pPIC9K and expressed in Pichia pastoris KM71 strain . After induction by methanol, HCB was expressed and secreted into the medium. The molecular weight of the recombinant procathepsin B was determined as about 38 kDa. The expressed product was confirmed to be HCB by immunoblotting assay using specific rabbit anti-HCB polyclonal antibody. The activity of the product was assayed by in situ hydrolysis (gelatin-SDS-PAGE). These results showed that HCB with proteolytic activity was expressed in P. pastoris KM71. This proenzyme can be used for further research on the activation of the proenzyme or industrial production.

The effects of adrenomedullin (ADM) on the L-type calcium currents (I(Ca,L)) and the mechanism of the signal transduction process were studied. Enzymatically isolated guinea-pig ventricular myocytes were used to measure ICa,L with whole-cell patch-clamp techniques. ADM at the concentrations of 1-100 nmol/L decreased ICa,L in a dose-dependent manner (P<0.05). ADM22-52) (100 nmol/L), a specific ADM-receptor antagonist, completely abolished the ADM-induced inhibition of ICa,L. Pretreatment of the cells with H-89 (10 micromol/L), a specific PKA inhibitor, did not attenuate the effects of ADM. Intracellular application of 10 micromol/L PKC19-36), a specific PKC inhibitor, prevented the ADM-induced inhibition of the ICa,L, while the specific PKC activator PMA could mimic the effects of ADM on the ICa,L. PMA (1 micromol/L) decreased the ICa,L by 32.26+/-4.20%(P<0.05). These findings indicate that ADM can inhibit the ICa,L in guinea-pig ventricular myocytes, and the inhibition is mediated by the specific ADM-receptor and an activation of protein kinase C.
Adrenomedullin ; pharmacology ; Animals ; Calcium Channels, L-Type ; metabolism ; Guinea Pigs ; Heart Ventricles ; cytology ; Myocytes, Cardiac ; drug effects ; metabolism ; Patch-Clamp Techniques ; Protein Kinase C ; metabolism

Objective To observe the urodynamic change after spinal cord injury at different levels and the relationship with neurogenic dysfunction of bladder and urethra. Methods Eighty female rats were divided into a control group (20 rats) , a suprasacral spinal cord injury group (30 rats) and a sacral spinal cord injury (30 rats). The urodynamic exam was performed with all the rats before and 20 days after the spinal cord injury model was established by surgical operation. Results The maximum bladder volume and compliance in the su- prasacral injury group were significantly less than the sacral spinal cord injury group and the control, the maxi- mum volume and compliance in sacral spinal cord injury group were significantly less than the control. The DLPP in suprasacral injury group was significantly higher than that in the sacral spinal cord injury group and the con- trol, the DLPP in sacral spinal cord injury group was significantly less than that in the control group. Conclu- sion Urodynamic study is very useful for the early diagnosis and individualized treatment of the neurogenic bladder after spinal cord injury.

Previously, we have reported that CKAP2 is involved in the maintenance of centrosome integrity, thus allowing for proper mitosis in primary hepatocytes. To understand this biological process, we identified the mitosis-specific phosphorylation sites in mouse CKAP2 and investigated CKAP’s possible role in cell cycle progression. Because we observed mouse CKAP2 depletion in amplified centrosomes and aberrant chromosomal segregation, which was rescued by ectopic expression of wild-type CKAP2, we focused on the centrosome duplication process among the various aspects of the cell cycle. Among the identified phosphorylation sites, T603 and possibly S608 were phosphorylated by CDK1–cyclin B1 during mitosis, and the ectopic expression of both T603A and S608A mutants was unable to restore the centrosomal abnormalities in CKAP2-depleted cells. These results indicated that the phosphorylation status of CKAP2 during mitosis is critical for controlling both centrosome biogenesis and bipolar spindle formation.
Animals ; Biological Processes ; Cell Cycle ; Centrosome* ; Ectopic Gene Expression ; Hepatocytes ; Mice ; Mitosis ; Phosphorylation*

OBJECTIVETo verify the effects of treatment with blood transfusion and scopolamine on severe chlorphenamidine poisoning (SCP).

METHODS400 patients with severe oral chlorphenamidine poisoning were randomly divided into two groups. 200 patients (Group I) were treated with the traditional combined therapy including gastrolavage, purgation and taking redox agent (methylene blue and vitamin C) while the other 200 patients (Group II) in addition to the above mentioned therapy, received blood transfusion and scopolamine injection.

RESULTSThe cure rate of Group II was 99.5% and significantly higher than that of Group I (91.0%, P < 0.01). The average time of improving in health in Group II [(8.71 +/- 1.49) h] was obviously shorter than those in Group I [(10.65 +/- 1.72) h, P < 0.01]. Blood methemoglobin concentrations in Group II at 3, 7, 12, 24 h after admission [(43.58 +/- 2.69), (34.21 +/- 2.30), (20.60 +/- 4.03), (13.50 +/- 1.65) g/L respectively] were obviously lower than those in Group I [(54.42 +/- 12.79), (42.17 +/- 22.34), (30.66 +/- 17.67), (19.01 +/- 0.61) g/L respectively, P < 0.01].

CONCLUSIONBlood transfusion and scopolamine had distinctive therapeutic effect on SCP to makeup the deficiency of redox agent. Combination of three therapies may potentiate the detoxication for chlorphenamidine.

Adult ; Antioxidants ; therapeutic use ; Blood Transfusion ; Chlorphenamidine ; poisoning ; Drug Therapy, Combination ; Female ; Humans ; Insecticides ; poisoning ; Male ; Mydriatics ; therapeutic use ; Poisoning ; therapy ; Scopolamine Hydrobromide ; therapeutic use ; Treatment Outcome

OBJECTIVETo express and purify the fusion protein of extracellular domain of human Ig domain-containing, neurite outgrowth inhibitor (Nogo) receptor-interacting protein-1 (LINGO-1(aa76-319)) in prokaryotic cells and prepare the rabbit anti-LINGO-1 polyclonal antibody (pAb).

METHODSThe 732 bp DNA sequence of hLINGO-1(aa76-319) was obtained from pCMV-SPORT6 by PCR and inserted into pET30a(+) plasmid to construct the prokaryotic expression plasmid pET30a(+)-hLINGO-1(aa76-319), which was subsequently transformed into E.coli. The target fusion protein was expressed with IPTG induction and purified by Ni(2+)-NTA affinity chromatography column. The antiserum against hLINGO-1(aa76-319) was obtained from the rabbits immunized with hLINGO-1(aa76-319), and the titer of the pAb was determined using enzyme linked immunosorbent assay (ELISA) and its specificity identified using Western blotting.

RESULTSThe prokaryotic expression plasmid pET30a(+)-hLINGO-1(aa76-319) was constructed successfully. Efficient expression of the target fusion protein was achieved with IPTG induction at the optimal concentration of 0.4 mmol/L and culture temperature at 37 degrees celsius; for 2.5 h. The hLINGO-1(aa76-319) fusion protein was effectively expressed in E.coli as inclusion bodies, and the soluble protein was obtained through denaturation and refolding procedures, and the purified fusion protein showed a purity above 90%. The titer of the anti-hLINGO-1(aa76-319) pAb obtained by immunizing the rabbits with the purified protein reached 1:1.6x10(6), and Western blotting confirmed its good specificity.

CONCLUSIONThe fusion protein hLINGO-1(aa76-319) with high purity has been obtained and the anti-hLINGO-1(aa76-319) pAb obtained shows a high titer and good specificity, which provide important experimental basis for further functional investigation of LINGO-1.

Animals ; Antibodies ; immunology ; isolation & purification ; Antibody Specificity ; Escherichia coli ; genetics ; metabolism ; Humans ; Immune Sera ; immunology ; Membrane Proteins ; biosynthesis ; genetics ; immunology ; Nerve Tissue Proteins ; biosynthesis ; genetics ; immunology ; Plasmids ; genetics ; Rabbits ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; immunology