AIM: To study the effect of double gene transduction mediated by lentiviral vectors on the characteristics of human embryonic stem cells (hESCs). METHODS: Using the backbone from inducibal dual and excisable transgene vector (iDuet101) , lentiviral vectors overexpressing cytotoxic T lymphocyte - associated molecule - 4 immuno-globulin (iDuet101 - CTLA4Ig) , indoleamine 2, 3 dioxygenase (iDuet101 - IDO) , and ubiquitin - C promoter - lueifer-ase - ires - puromycin ( ULIP) were constructed and packaged according to the standard recombinant techniques. Human embryonic stem cells (hESCs) were first transduced with iDuet101, iDuet101 - CTLA4Ig or iDuet101 - IDO, then, after the selection, were transduced again with ULIP. The expression and function of the exogenous genes were detected. Immu-nohistochemistry, RT - PCR and flow cytometry were applied for detection of embryoid bodies ( EB) formation in vitro and in vivo teratoma formation. RESULTS: Double - transduced hESCs showed typical shape of cell clones and positive staining of tumor rejection antigen -1 - 60 ( Tra -1 - 60 ) and octomer transcription factor - 4 ( OCT - 4 ). The formation of EB was observed, in which a - fetoprotein (AFP), paired box gene 6 ( Pax6) and Musashi 1 ( MSI1) were positively expressed. The cells formed teratomas, and the luciferase signals existed until 28 days after xeno - transplantation. CONCLUSION : Double transduction of non - transcriptional factors mediated by lentiviral vectors does not affect the cell growth rate and their differentiation ability.

Objective To observe the efficacy of pregabalin in the treatment of painful diabeticperipheral neuropathy.Methods In a 8-week study,50 patients were randomly assigned into two groups:the therapy group and the control group.The patients in the therapy group were treated with pregabalin 150 mg/d,and the patients in the control group were treated with carbamazepine 200 mg/d.The dose of pregabalin was increased to 300 mg/d till pain was alleviated in the therapy group,and the dose of carbamazepine was increased to 600 mg/d till pain was alleviated in the control group.The therapeutic effect and VAS score in the two groups were observed and evaluated.Results After 1 and 8 weeks' treatment,the therapeutic effect of therapy group were significantly superior to the control group ( At 1 week:U =2.028,P =0.046 ; At 8 week:U =3.540,P ＜0.001 ).After 1 and 8 weeks' treatment,the VAS scores of therapy group were significantly lower than the control group ( At 1 week:5.13 ± 1.76 vs.6.74 ± 1.52,P ＜ 0.05 ; At 8 week:1.13 ± 0.45 vs 3.27 ± 1.04,P ＜ 0.05).There were 5 patients (20％) and 10 patients (40％) had adverse reaction in the therapy and the control groups respectively.Conclusion Pregabalin is effective and safe for the treatment of painful diabetic peripheral neuropathy.

Objective To investigate the changes of brain volumes in neuromyelitis optica (NMO)patients using voxel-based morphometry (VBM) method, and preliminarily explore the pattern of cerebral anatomical impairment. Methods Twenty-three clinically defined NMO patients and 15 gender and age matched healthy volunteers underwent 3-dimensional (3D) fast spoiled gradient echo (FSPGR) sequence scanning on 3.0 Tesla MR systen. Raw data was processed and analyzed using statistical parametric mapping (SPM) 5. Whole brain volumes included grey matter volume (GMV), white matter volume (WMV), total intracranial volume (TIV), grey matter fraction (GMF), white matter fraction (WMF),brain tissue fraction (BTF) and regional brain volumes between the two groups were compared by independent samples t-test and an Pearson were performed to compare the regional brain volumes and the ages. Results GMV of NMO group[(610. 2 ± 55.0) ml] was significantly decreased comparing to healthy control group[(657. 2 ± 36. 3) ml] (t = - 2. 915, P ＜ 0. 05). The age of NMO patients [(40 ± 9) years old] showed negative correlation with GMF [(42. 5 ± 2. 6) %] (r = - 0. 673, P ＜ 0. 05). Regional brain volume analysis showed decreased GMV in left insula and bilateral posterior cingutates in NMO patients,while decreased WMV was found in left frontal and left parietal white matter. Conclusion VBM could detect brain volume changes sensitively. Total grey matter volume in NMO patients was decreased comparing to HC group. Regional grey matter atrophy in NMO patients occurred in left insular and bilateral posterior cingutates, regional white matter atrophy occurred in left frontal and left parietal lobe.

A reverse transcription polymerase chain reaction (RT-PCR) and single-strand conformation polymorphisms (SSCP) assay were applied to rapidly detect the molecular variability in CP and SP genes among seven isolates of Rice stripe virus in China. The PCR results showed that the CP gene of JD isolate and SP gene of PJ isolate could not be amplified. SSCP analysis showed that there were completely different electrophoretic pattern of CP gene among six isolates. To SP gene, SSCP results also discovered polymorphisms. There were five patterns among these isolates, and the pattern of YL and BS isolates were same.

OBJECTIVETo establish a method for studying efficacious materials of traditional Chinese medicines from an overall perspective.

METHODCarthamus tinctorius was taken the example. Its major components were depleted by preparing liquid chromatography. Afterwards, the samples with major components depleted were evaluated for their antioxidant effect, so as to compare and analyze the major efficacious materials of C. tinctorius with antioxidant activity and the contributions.

RESULTSeven major components were depleted from C. tinctorius samples, and six of them were identified with MS data and control comparison. After all of the samples including depleted materials are compared and evaluated for their antioxidant effect, the findings showed that hydroxysafflor yellow A, anhydrosafflor yellow B and 6-hydroxykaempferol-3, 6-di-O-glucoside-7-O-glucuronide were the major efficacious materials.

CONCLUSIONThis study explored a novel and effective method for studying efficacious materials of traditional Chinese medicines. Through this method, we could explain the direct and indirect contributions of different components to the efficacy of traditional Chinese medicines, and make the efficacious material expression of traditional Chinese medicines clearer.

Alkalies ; chemistry ; Antioxidants ; chemistry ; Carthamus tinctorius ; chemistry ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; Glucosides ; chemistry ; Mass Spectrometry

P-selectin, one of the membrane proteins, expresses on platelet and endothelia and interacts with P-selectin glycoprotein ligand-1 (PSGL-1) on leukocyte membrane. This interaction mediates leukocytes rolling on endothelial membrane and then induces leukocyte recruitment to the site of infection or tissue injury. In the present study, we constructed the recombinant wild type human P-selectin, its calcium-binding sites mutants and recombinant PSGL-1-globulin (PSGL-1-Rg). They expressed in Sf9 cells by using the baculovirus expression system and were purified by TalonTM metal or Protein A affinity chromatography. The results showed that the recombinant PSGL-1-Rg interacted with recombinant wild type P-selectin and two P-selectin mutants with 2 calcium-binding sites mutation respectively, but could not bind to the P-selectin mutant with all 4 calcium-binding sites mutation. Therefore, we verified the importance of P-selectin calcium-binding sites for its interaction with PSGL-1.
Binding Sites ; Calcium ; metabolism ; Humans ; Leukocytes ; metabolism ; Membrane Glycoproteins ; metabolism ; Mutation ; P-Selectin ; metabolism ; Recombinant Proteins ; metabolism

Antigens ; chemistry ; Fixatives ; chemistry ; Humans ; Hydrogen-Ion Concentration ; Immunohistochemistry ; methods ; Microwaves ; Staining and Labeling ; methods

Objective To observe the preventive effect of Anduolin(ADL) on radiation-induced lung injury in mice.Methods Totally 180 of Kunming mice were randomly classified into six groups:normal control group ( N ),irradiation control group ( R),irradiation plus low dose ADL group ( L),irradiation plus middle dose ADL group ( M),irradiation plus high dose ADL group ( H),and irradiation plus Dexamethasone group (D).The mice except group N were irradiated with 20 Gy of 6 MV X-rays on whole lung.The mice in group L,M and H were given with ADL 1 d before irradiation and continued for 6 weeks after irradiation.At 2,4 and 6 weeks after irradiation,the general situation and the lung pathological changes of mice were observed.The lung wet weight,collagen contents of the whole lung tissue,hydroxproline concentration,and TGF-β1 expression in the lung were also delected.Results Compared to the group R,the mice breathing rate,hydroxproline concentration,and TGF-β1 expression in the group L were not significantly changed.While in the groups M,H and D,the breathing rate,the generation of hydroxproline and the expression of TGF-β1 were decreased significantly ( F =2.668-161.646,P ＜0.05).In addition,ADL alleviated the pathological changes on radiation-induced lung injury in mice.Conclusions ADL might have the preventive effect on radiation-induced lung injury in mice.

OBJECTIVETo explore the application of sinusitis mixture (SM) in endoscopic sinussurgery, thereby improving clinical curative rate of chronic sinusitis and nasal polyps.

METHODSA totalof 50 chronic sinusitis patients were equally assigned to the experimental group (nasal douching by SM)and the control group (nasal douching by Compound Sodium Chloride Injection). Mucosa tissue 0.1 cmbefore natural opening was collected before surgery, at week 4, 12, and 24 after surgery. Changes ofmucosa cilia cells, goblet cells, stroma of mucosal membrane, inflammatory cells, and mucous glandwere observed. The numbers of goblet cells in the upper epithelia and ciliated cells, as well as their ratioswere calculated.

RESULTSThere was statistical difference in cavity cleaning time, cavity mucosal epithelization time, numbers of goblet cells in the upper epithelia and ciliated cells, as well as their ratio between the two groups (t = -2.342, -2.015, -2.145, respectively; P < 0.05).

CONCLUSIONSM could effectively promote and accelerate cleaning and mucosal epithelization of functional endoscopic sinus surgery, and significantly promote mucosal ciliary structure and function recovery of ostium-meatus nasicomplex.

Chronic Disease ; Drugs, Chinese Herbal ; therapeutic use ; Endoscopy ; Epithelium ; pathology ; Humans ; Mucous Membrane ; cytology ; pathology ; Sinusitis ; surgery

Objective To establish a new model of cardiac arrest (CA) in rats by transcutaneous electrical epicardium stimulation. Method Two acupuncture needles connected to the anode and cathode of a stimulator were transcutaneously inserted into the epicardium as electrodes. The stimulating current was steered to the epicardium and the stimulation was maintained for 3 minutes to induce CA. Cardiopulmonary resuscitation (CPR) was performed at 6 minutes after a period of nonintervention. Results The success rate of induction was 12/20 at the current intensity of 1 mA; and reached 20/20 when the current intensity was increased to 2 mA. The average time from the electrical stimulation to CA induction was (5. 10 ± 2. 81) seconds. When the electrical stimulation stopped, 18/20 rats had ventricular fibrillation and 2/20 rats had pulseless electrical activity. CPR was performed for averagely 207.4 ( ± 148.8) seconds. The restoration of spontaneous circulation was 20/20. The death rate within 4 hours after CA was 5/20, and the 72-hour survival rate was 10/20. There were only two cases of complications, a minor muscle contraction and a minor lung lobe injury. Conclusions The model of CA in rats induced by transcutaneous electrical epicardium stimulation is a stable model that requires low-intensity current and has fewer complications.