Bel1, a transactivator of prototype foamy virus (PFV), plays pivotal roles in the replication of PFV. Previous studies have shown that Bel1 bears a nuclear localization signal (NLS), but its amino acid sequence remains unclear and the corresponding importins have not been identified. In this report, we inserted various fragments of Bel1 into an EGFP-GST fusion protein and investigated their subcellular localization by fluorescence microscopy. We found that the 215PRQKRPR221 fragment could direct nuclear localization, which accords with the consensus sequence K(K/R)X(K/R) of monopartite NLS. Point mutation experiments revealed that K218, R219, and R221 are essential for the nuclear localization of Bel1. The results of the GST-pulldown showed that the Bel1 fragment with residues 215-223, which bears the NLS, interacts with KPNA1, KPNA6, and KPNA7. This result suggests that KPNA1, KPNA6, and KPNA7 maybe involved in Bel1 nuclear translocation.
Cell Line ; Cell Nucleus ; genetics ; metabolism ; virology ; Humans ; Nuclear Localization Signals ; genetics ; metabolism ; Protein Binding ; Protein Transport ; Retroviridae Infections ; genetics ; metabolism ; virology ; Retroviridae Proteins ; chemistry ; genetics ; metabolism ; Spumavirus ; chemistry ; genetics ; physiology ; Trans-Activators ; chemistry ; genetics ; metabolism ; alpha Karyopherins ; genetics ; metabolism

No abstract available.
Humans ; Mucinoses

AIMTo synthesize four water-soluble metal porphyrins [5, 10, 15, 20-tetra[4-(4'-pyridine-1) butyloxy phenyl] metalloporphyrins bromide, metal = Zn (I), Cu (II), Mn (III) and Co (IV)] as analogous enzyme having two anti-active oxygen functions.

METHODSThe first function, scavenging O2-, has been proved by using riboflavine-methionine photoreduction methods. The second function, scavenging H2O2, has been demonstrated by using the oxidating Vit C. The third function, scavenging HO*, has been demonstrated by using Fenton reaction. The complexes were measured by the mice liver homogenate technique of mice.

RESULTSFour model compounds could scavenge O2- in the concentration range of 1.0 x 10(-5) - 1.0 x 10(-6) mol x L(-1), decompose H2O2 in the concentration of 1.5 x 10(-6) - 1.0 x 10(-6) mol x L(-1), scavenge HO* in the concentration of 2.0 x 10(-8) - 1.0 x 10(-8) mol x L(-1). All showed that they had obvious action of decreasing the lipid peroxidation in the concentration of 1.0 x 10(-7) mol x L(-1).

CONCLUSIONAll above-mentioned complexes were considered to be qualified analogous enzymes of anti-active oxygen.

Animals ; Cobalt ; Copper ; Free Radical Scavengers ; chemical synthesis ; pharmacology ; Hydrogen Peroxide ; metabolism ; Hydroxyl Radical ; metabolism ; In Vitro Techniques ; Lipid Peroxidation ; drug effects ; Liver ; metabolism ; Malondialdehyde ; metabolism ; Manganese ; Metalloporphyrins ; chemical synthesis ; pharmacology ; Mice ; Reactive Oxygen Species ; metabolism ; Zinc

OBJECTIVETo study the role of proximal nerve stimulation at Erb point in the early diagnosis of Guillain-Barré syndrome (GBS) in children.

METHODSThirty-two children who were diagnosed with GBS between October 2013 and December 2014 received neurophysiological examination. Thirty healthy children were used as controls. Compound muscle action potentials and distal motor latency of the median and ulnar nerves were determined and analyzed after nerve stimulation at the wrist, elbow, and Erb point in the two groups. Moreover, F-wave latency of the median nerve and H-reflex latency of the tibial nerve were measured and analyzed in the two groups.

RESULTSThe F-wave and H-reflex latencies were significantly longer in the patient group than in the control group (P<0.05). In thirty-two patients, the numbers of patients with abnormal amplitude, abnormal latency, and conduction block at Erb's point were 24 (75%), 22 (69%), and 20 (62%), respectively. The patient group had significantly lower amplitudes but significantly longer latencies of the ulnar and median nerves at Erb point than the control group (P<0.05). There were no significant differences in the amplitudes and latencies at the wrist and elbow between the two groups (P>0.05).

CONCLUSIONSThe nerve stimulation at Erb point holds promise as a routine examination for the early diagnosis of GBS.

Adolescent ; Child ; Early Diagnosis ; Electrodiagnosis ; methods ; Female ; Guillain-Barre Syndrome ; diagnosis ; physiopathology ; H-Reflex ; Humans ; Male ; Neural Conduction ; physiology ; Reaction Time

Objective To analyze the recurrence and metastasis in patients with non-small cell lung cancer (NSCLC) after minimally invasive surgery. Methods 123 patients with NSCLC underwent thoracoscopic lobectomy from January 2008 to December 2013 were enrolled in the study. Their perioperative data and follow-up results were analyzed, and postoperative recurrence and metastasis were recorded. Multivariate logistics regression analysis was performed to investigate the influencing factors of postoperative recurrence and metastasis. Results The median operative time was 165 min (60 ~ 430 min) and the median intraoperative blood loss was 95 ml (20 ~ 3100 ml). Postoperative complications occurred in 15 cases (12.2%). All patients were followed up regularly after discharge, and the median follow-up time was 23.5 months (6 ~ 69 months). During the follow-up period, postoperative recurrence and metastasis occurred in 36 cases (29.3%) and 42 cases (34.1%), including 16 cases (13.0%) patients simultaneously appeared recurrence and metastasis. The ipsilateral lung (52.8%) was the most common site of recurrence, followed by mediastinal lymph nodes (38.9%). Bone (28.6%) was the most common site of metastasis,followed by contralateral lung (26.2%) and brain (19.0%). Multivariate logistic regression analysis showed that stage Ⅱ-Ⅲ, mediastinal lymph node metastasis and low differentiation were independent risk factors of postoperative recurrence (P < 0.05) while stage Ⅱ ~ Ⅲ , number of lymph node metastasis ≥ 3, without postoperative radiotherapy and chemotherapy were independent risk factors of postoperative metastasis (P < 0.05). Conclusions For patients with NSCLC, recurrence and metastasis after thoracoscopic lobectomy occurred mainly in the ipsilateral lung and bone. TNM staging, mediastinal lymph node metastasis, differentiation degree, lymph node metastases, postoperative radiotherapy and chemotherapy were related with postoperative recurrence and metastasis.

Cajanus cajan L. is a natural plant, which contains a lot of potential active components. In the present study, we identified the effects of the stilbene extract from Cajanus cajan L. (sECC) on hepatic cholesterol metabolism in diet-induced (for 4 weeks) hyperlipidemic Kunming mice. All experimental mice were divided into 5 groups: control group, high lipid model group, sECC-treated with 200 or 100 mg kg(-1), and simvastatin (Sim, 12 mg kg(-1)) treated group. The mice were fed with fat and cholesterol-enriched chow except control mice that were fed with standard diet. The effects of sECC were investigated by monitoring serum and liver lipid profile (i. e. cholesterol homeostasis) in mice. To further explore the mechanism of sECC, hepatic cholesterol 7alpha-hydroxylase (CYP7A1) and low density lipoprotein (LDL) receptor expressions in cholesterol homeostasis were analyzed by reverse transcription PCR. After 4 weeks pretreatment, the mice in the high lipid model group showed markedly higher serum and hepatic lipid contents than control group (P< 0.01). Compared with high lipid model group, the increased serum and hepatic lipid contents were markedly attenuated by sECC (200 mg kg(-1)), the serum and hepatic total cholesterol were reduced by 31.5% and 22.7% (P<0.05), respectively. The triglyceride contents of serum and liver were also lowered by 23.0% and 14.4%, respectively. At the same times, serum LDL cholesterol decreased by 53.0% (P<0.01). The mRNA expressions of hepatic CYP7A1 and LDL-receptor were significantly enhanced in the mice administered with sECC (200 mg kg(-1)), whereas those expressions were suppressed by the fat and cholesterol-enriched diet. These data indicate that sECC reduces the atherogenic properties of dietary cholesterol in mice. It is indicated that expression enhancement of hepatic LDL-receptor and cholesterol 7alpha-hydroxylase may be responsible for the hypercholesterolemic effect.
Animals ; Anticholesteremic Agents ; isolation & purification ; pharmacology ; Body Weight ; drug effects ; Cajanus ; chemistry ; Cholesterol ; blood ; metabolism ; Cholesterol 7-alpha-Hydroxylase ; biosynthesis ; genetics ; Cholesterol, LDL ; blood ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Gene Expression Regulation ; Hypercholesterolemia ; blood ; genetics ; metabolism ; pathology ; Liver ; metabolism ; pathology ; Male ; Mice ; Organ Size ; drug effects ; Plant Leaves ; chemistry ; Plants, Medicinal ; chemistry ; RNA, Messenger ; metabolism ; Receptors, LDL ; biosynthesis ; genetics ; Stilbenes ; isolation & purification ; pharmacology ; Triglycerides ; blood ; metabolism

Objective To improve vibration resistance of conventional blood transportation kits and mitigate hemolysis during transportation.Methods The structure of a blood transportation kit was modified.We installed a suspension brac-kets within the kit,added buffer material between the brackets,and tested the vibration-suppressing effect compared with the conventional blood transportation kit.Results Rubber and plastic materials between brackets were added,and double membrane suspension brackets were installed.After 4 and 6 min of vibration,free hemoglobin(FHb)[(1559.7 ±1038.5) and(1886.2 ±1023.8)mg/L],lactic dehydrogenase levels[(135.3 ±67.7)and(195.7 ±123.6)U/L]and hemolysis rate[(0.35 ±0.34)%and(0.42 ±0.38)%]in the conventional transportation kit were significantly higher than in the vibration-suppressing kit.K+did not change significantly,and was comparable in both groups at each time point.After 4 and 6 min of vibration, FHb in the conventional transportation kit exceeded the standard.However, after 12 min of vibration,FHb[(560.1 ±342.3)mg/L]in the vibration-suppressing kit were within the standard range.No bacterial growth was detected in either group.Conclusion The vibration-suppressing kit under research shows a better 1986vibration-suppressing effect,which could improve blood support capability in case of emergency.

Objective To study the effect of ulinastatin on no reflow or slow flow in the infarct related artery in patient with acute ST-elevation myocardial infarction (STEMI) undergoing emergency percutaneous coronary interventional therapy (PCI).Methods 180 STEMI patients were divided into the control group (n=100) and the ulinastatin treatment group (n=80).The control group received conventional PCI treatment and the treatment group received conventional PCI treatment plus ulinastatin. The level of serum inflammatory factors IL-6,IL-10,superoxide converting enzyme,the infarct related coronary artery reperfusion TIMI flow grade (TFG) and myocardial perfusion grade (TMPG),the results of postoperative cardiac ultrasound examination and clinical outcomes were compared between the two groups.Results Compared with the control group,the level of IL-6 was decreased,while the levels of IL-10 and superoxide converting enzyme were increased significantly in the ulinastatin treatment group(P<0.05).The TFG and TMPG of the infarct related vessels were increased significantly in the ulinastatin treatment group. The left ventricular end diastolic diameter[(54.6 ± 5.2 mm vs. (50.4±4.6) mm,P=0.046)]and left ventricular ejection fraction [(58.4±10.2) % vs. (62.2±9.8) % P=0.048] showed statistical difference between the two groups.Compared to the control,the major cardiovascular event rate of the treatment group during hospitalization (1% vs. 5%, P=0.038), after one month (1.2% vs. 3%,P=0.046) and 6 months (3% vs 12%,P=0.018) were all significantly lower .There was no significant difference in mortality between the 2 groups.Conclusion Ulinastatin may lower the incidence of no flow and slow flow after emergency PCI,improve heart function and the lower the rates of MACE.

OBJECTIVETo investigate the possible pathways for ovarian injury after administration of cyclophosphamide in rats.

METHODSAdult SD rats received a single injection of saline vehicle or chemotherapeutic agent cyclophosphamide, and 8 weeks later, the ovaries were removed, fixed and serially sectioned for pathological examination and ovarian follicle counting. The expression of stem cell factor (SCF) protein was evaluated by immunohistochemistry and immunoreactive score, and SCF mRNA expression determined by RT-PCR in rat ovaries.

RESULTSCyclophosphamide had a detrimental effect on ovarian stromal function and lead to primordial follicle loss. Immunoreactive SCF antigens were expressed on the oocytes in the primordial and primary follicles of rat ovaries, and also in the granulosa cells of the secondary follicles and early antral follicles. There was a higher granulosa SCF, lower oocyte SCF and higher SCF mRNA level in the ovaries of the rats exposed to cyclophosphamide as compared with those in control rat ovaries (P <0.05).

CONCLUSIONAltered SCF expression in the ovaries of rats exposed to cyclophosphamide can be helpful for understanding the mechanisms for chemotherapeutic drug-induced ovarian damage.

Animals ; Cyclophosphamide ; adverse effects ; Female ; Gene Expression ; drug effects ; Granulosa Cells ; drug effects ; metabolism ; Oocytes ; drug effects ; metabolism ; Ovary ; cytology ; drug effects ; injuries ; metabolism ; Rats ; Rats, Sprague-Dawley ; Stem Cell Factor ; genetics ; metabolism

OBJECTIVETo prepare eukaryotic expression of rotavirus (RV) SA11 capsid protein VP7, and to generate and purify yolk immunoglobulin (IgY) antibodies against the recombinant VP7 from Roman hens.

METHODSMA104 cells were infected with the standard SA11 strain and the culture fluid was collected. A DNA fragment of 978 bp encoding SA11 VP7 was obtained by RT-PCR amplification from genomic RNA of RV SA11. The PCR products were ligated to pMD18-T vector following the confirmation by DNA sequencing and sub-cloned into pPICZalphaB. The recombinant pPICZalphaB-SA11 VP7 was transformed into E coli Top10. The plasmids were linearized by digestion of BstXI and transformed into Pichia pastoris X-33 through electroporation by DNA sequencing. The transformants were induced with methanol for expression. The cultural supernatant was subjected to SDS-PAGE and Western blotting. Fusion expression was purified through the column of affinity chromatography. IgY was identified and purified by SDS-PAGE and Western blotting from eggs of Roman hens immunized with recombinant SA11 VP7.

RESULTSThe RNA extracted from the RV culture fluid consisted of 11 bands visualized by silver staining. The expression vector pPICZalphaB-SA11 VP7 was constructed and the fusion protein in Pichia pastoris X-33 was harvested and purified. The recombinant SA11 VP7 with molecular weight of 40 200 was identified by Western blotting. The IgY antibodies against the recombinant SA11 VP7 were produced with a purity of 95 percent and yield of 10.2 mg per egg.

CONCLUSIONThe preparation of IgY antibodies to recombinant SA11 VP7 might lay a foundation for the development of vaccines and diagnostic techniques.

Animals ; Antigens, Viral ; genetics ; immunology ; metabolism ; Capsid Proteins ; genetics ; immunology ; metabolism ; Chickens ; Cloning, Molecular ; Immunoglobulins ; immunology ; isolation & purification ; Recombinant Proteins ; genetics ; immunology ; metabolism