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 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.

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.

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.

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

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

Coptidis Rhizoma-Euodiae Fructus has been widely used for the treatment of digestive diseases since Song Dynasty, and therapeutic efficacy is very obvious. Modern research found that alkaloids are the main bio-active constituents, and some of their contents have striking difference after compatibility of the two herbs. The Chinese medicine pair (CMP) has extensive biological activities, such as the effect of gastrointestinal effect, anti-tumor, lowering the blood pressure and blood fat and so on. And some action mechanism of CMP also got partial demonstration. This paper mainly summarized the bio-active constituents, compatibility effects, action mechanism and clinical applications of the CMP, which can provide a basis for further research and development of the CMP.
Animals ; Drug Interactions ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Evodia ; chemistry ; Humans ; Medicine, Chinese Traditional ; methods

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

OBJECTIVEIn this study, five heavy metals contamination of soil and different parts of Panax notoginseng in the plantation area was investigated. Analysis of heavy metals correlation between the planting soil and P. notoginseng; and the absorption and accumulation characteristics and translocation of soil heavy metals by P. notoginseng plants was revealed.

METHODThrough field investigation and laboratory analytical methods, analysis of China's 30 different soil P. notoginseng origin and content of heavy metals in five different parts of the P. notoginseng plant content of heavy metals.

RESULTThe results revealed that the soil heavy metals should not be neglected in the plantation area Referring to the national soil quality standards (GB15608-1995), the excessive degree of soil heavy metals pollution showed Hg > As > Cd > Cr in the plantation area, and Pb content of soil was in the scope of the standard. Refer to 'Green Industry Standards for Import and Export of Medical Plants and Preparations', the excessive degree of heavy metals content of P. notoginseng plants showed As > Pb > Cr > Cd, and Hg content of plants was in the scope of the standard. Concentrations of five heavy metals of underground parts of P. notoginseng plants are higher than aboveground, and heavy metals elements are more concentrated in the root, followed by the rhizome of P. notoginseng plants. Heavy metal accumulation characteristics of the different parts of the P. notoginseng of the overall performance is the root > the rhizome > the root tuber > leaves > stems. From the point of view BCF value analysis of various parts of the P. notoginseng plants to absorb heavy metals in soil, BCF values of all samples were less than 1, description P. notoginseng not belong Hyperaccumulator. From the view of transportation and related analysis of the soil-P. notoginseng systems, the rhizome of P. notoginseng and the content of As and Cr in soil was significantly correlated, the root of P. notoginseng and the content of Cd in soil was significantly correlated, and no significant correlation between the other indicators. Through the analysis of transportation transfer coefficient showed: Pb, As and Cr are not easy to transport aboveground part from the underground, but Cd and Hg are relatively easy to transport stems from rhizome, the migration of five heavy metals in the aerial part is relatively strong, and heavy metal of stems is easily transported to the leaves.

CONCLUSIONP. notoginseng does not belong to the enrichment of heavy metals in crops, especially for Hg in soil with strong patience. In survey area, the content of heavy metals of P. notoginseng's planting soil is relatively high, and the heavy metals As, Pb, Cr, Cd of P. notoginseng also exist heavy metals exceeded problems. Due to the presence of heavy metals in crops internal absorption and translocation of special laws, accumulation of heavy metals varied significantly in different parts of P. notoginseng. The overall, the performance for the heavy metal content of the underground parts is more than aboveground, it explain heavy metals of P. notoginseng plants is still the main source of the soiL Therefore, the key to control of planting area soil environmental quality and reduce exogenous harmful substances secondary pollution of soil in the cultivation process are to study and solve the heavy metals pollution problem of P. notoginseng.

Adsorption ; China ; Laboratories ; Metals, Heavy ; analysis ; Panax notoginseng ; chemistry ; Soil ; chemistry ; Soil Pollutants ; analysis

OBJECTIVETo investigate the effect of oligochitosan in promoting intestinal absorption of protoberberine alkaloids in extracts from Corydalis saxicola total alkaloids.

METHODThe in vitro single-pass intestinal perfusion model in rats was established to study the changes in absorption kinetic parameters of dehydrocavidine, berberine hydrochloride and palmatine chloride in C. saxicola total alkaloids after the addition of different concentrations oligochitosan and evaluate the effect of oligochitosan in promoting intestinal absorption of the drugs.

RESULTThe concentration of oligochitosan had different effects on the absorption rate constant (Ka) and apparent permeability coefficient (Peff) of the three active component in rat intestines. Ka and Peff in 0.5% oligochitosan group significantly increased, indicating a stronger effect in promoting the absorption.

CONCLUSIONOligochitosan has a certain effect in promoting the intestinal absorptions of protoberberine alkaloids in C. saxicola total alkaloids.

Animals ; Berberine Alkaloids ; administration & dosage ; pharmacokinetics ; Chitin ; administration & dosage ; analogs & derivatives ; Corydalis ; chemistry ; Drugs, Chinese Herbal ; administration & dosage ; pharmacokinetics ; Intestinal Absorption ; drug effects ; Intestines ; drug effects ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley