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

Objective To assess the status and severity of bone disease in patients with multiple myeloma (MM) by using chest computerized tomography (CT) and the relationship between clinical prognostic parameters and bone disease.Methods All 46 newly diagnosed MM in-patients received both imaging tests of chest CT and plain X ray.An experienced radiologist reviewed all the imaging data.Clinical laboratory parameters,stages of Durie-Salmon (DS) and International Staging System (ISS) were evaluated.Five cytogenetic abnormalities of bone marrow myeloma cells were tested by fluorescence in situ hybridization (FISH).Results The sensitivity of CT and X ray to determine pathological fractures was comparable,the positive rates of which were 41.3％ (19/46) and 30.4％ (14/46) respectively (P =0.29).Nevertheless,the positive rate of osteolytic lesions ascertained by CT was significantly higher than that by X ray (P ＜ 0.001),60.9％ (28/46) vs 13.0％ (6/46) with diameter 5-10 mm and 50.0％ (23/46) vs 10.9％ (5/ 46) with diameter more than 10 mm.Osteolytic lesion numbers found by CT were more than those by X ray [5(0-21) vs0(0-4) lesions with diameter5-10 mm (P＜0.001),2(0-14) vs0(0-2) lesions with diameter more than 10 mm (P ＜ 0.001),respectively].Patients with positive osteolytic lesions had higher percentage of RB1 gene deletion[46.7％ (14/30) vs 18.8％ (3/16),P ＜0.001],D13s319 deletion [43.3％ (13/30) vs 18.8％ (3/16),P ＜0.001] and high risk cytogenetic abnormalities[50.0％ (15/30) vs 25.0％ (4/16),P ＜ 0.001].Conclusions Chest CT is more sensitive than plain X ray in detecting osteolytic myeloma bone disease.Osteolysis determined by CT is relevant to clinical DS stages and risk stratification of cytogenetic abnormalities.

Objective To investigate the effects of plaque staining test on periodontal health of middle-aged and elderly patients with dental implants. Methods Ninety patients with dental implants were included in the study and divided into test group (n=45) and control group (n=45) by using random digit table. In the test group, the patients were all instructed with both routine health education and dental plaque dyeing education, while the control group received routine health education only. The plaque index (PLI) and gingival index (GI) were assessed at months 1 and 6 after implant restoration. Results One month after dental implantation, there were no significant differences in PLI and GI between the two groups (P>0.05), while 6 months after the implantation, the differences in PLI and GI between them were significantly different ( P<0 . 01 ) . Conclusion The plaque dyeing technology applied in the oral health education to elderly patients with dental implants can reduce plaque index and gingival index so that their periodontal health is enhanced.

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.

Objective To investigate the relationship between the change of pre-ablative TSH after thyroid hormone withdrawal(THW) and the response of subsequent 131I therapy in patients with low to intermediate risk DTC after total or near total thyroidectomy.Methods A total of 120 DTC patients (38 males,82 females,age (40.8±10.9) years) were enrolled in this retrospective study.Serial TSH levels determined on the day of THW and on the day of receiving 131I ablative therapy were monitored,which were marked as TSH1 and TSH2 accordingly.The THW duration (t) was recorded,the change of TSH was defined as △TSH and the change rate of TSH was calculated (V=△TSH/t).The responses to 131I therapy were classified as excellent response (ER),indeterminate response (IDR),biochemical incomplete response (BIR) and structural incomplete response (SIR) according to ATA guideline.According to the TSH2(mU/L) levels,patients were divided into G1 group (30≤TSH2＜60),G2 group (60≤TSH2＜90),G3 group (90≤TSH2＜ 120),G4 group (120≤TSH2＜150) and G5 group (TSH2 ≥ 150).Clinical and pathological features,THW duration,the change rate of TSH,residual thyroid,131 I dose and follow-up time were compared among these groups.In order to evaluate the relationship between response to 131I ablation and change rate of TSH,patients were divided into V1 group (V≤2.5),V2 group (2.5＜V≤5.0) and V3 group (V＞5.0),and their responses to 131I ablation were compared.Patients were classified into RI group (including ER and IDR)and R2 group (including BIR and SIR),the differences of clinical and pathological features,131I doses between the two groups were explored.Furthermore,logistic regression was performed to identify factors associated with BIR and SIR.Results Patients with male gender (x2=11.863),younger age (F =4.975),and faster TSH change rate (H =44.911) and lower thyroid residue (H =18.159) achieved a higher value of TSH2(all P＜0.05).G3 group presented the highest rate of ER (83.8％,31/37).The percentage of ER + IDR in V2 group was higher than those in V1 group and V3 group,which was 92.4％ (61/66),85.7％ (18/21) and 5/7,respectively,but the difference was not significant (U=407.5,P＞0.05).TSH2 level (OR=0.835) and pre-ablative Tg level (OR =1.196) were independent factors in predicting BIR and SIR (both P＜0.05).Conclusions The changing rate of TSH before 131 I ablation may not be associated with the response to 131I therapy in patient with low to intermediate risk DTC,while the level of TSH2 does.Patients with TSH2 ranging from 90 to 120 mU/L could be of help in achieving a better clinical response.