Aim According to various target sites of HLA-E mRNA,to design and synthesize 3 pieces of HLA-E siRNA chain,to compare quantitatively their efficiency of silencing gene in BEL-7402 with HLA-E(+)in order to select the dominant siRNA.Methods The hepatocarcinomal BEL-7402 cells,induced by 5×10~5 IU·L~(-1) IFN-γ,expressed HLA-E(+) and was pured by flow cytometry selecting as target cells for research.3 pieces of specific siRNA(A,B,C group)were designed and chemically synthesized,then the concentration of which(0.1 mmol·L~(-1))was respectively transfected through Lipofectamin 2000 into target cells.After 48 h,the gene silent effect on HLA-E gene in A,B and C groups was quantitatively observed by cytoimmunofluorence,flow cytometery,Western blot and real-time PCR,as well as on NK cytotoxicity to target cells tested by NK killing rate.Results Compared with those of control or non-specific siRNA group,HLA-E antigen,protein product,HLA-E mRNA and HLA-E molecule on cell surface were statistically down-regulated in A,B,and C group(P<0.01),whose were silenced more (above 90%) in B or C group than in A group (P<0.01).The NK killing rate in A,B and C groups was dominantly improved(P<0.01),which in B or C group was higher than in A group (P<0.01).Conclusion The targeted siRNA can specifically and high-efficiently silence HLA-E expression in hepatocarcinomal cells,and may keep them from immunoescape through non-classic HLAⅠ pathway to imply new strategy for hepatocarcinomal gene-immunotherapy.

To provide accurate information on geographic distribution of crude drug Sailonggu in the plateau, we identified zokor species (Eospalax spp.) in Qinghai-Tibet Plateau using molecular methods. Based on the mitochondrial cytochrome B (cytb) gene sequences, we then extracted haplotypes from these sequences and reconstructed phylogenetic trees for the haplotypes using both maximum likelihood (ML) and Bayesian inference (BI) methods. Based on the trees, the species of each sample were determined. Five hundred and three samples from 35 populations were sequenced and their whole cytb sequences (1140 bp) were obtained. From these sequences 150 haplotypes were detected, in which, 126 were Eospalax baileyi, 20 were E. cansus, and 4 were E. smithi of the 35 populations, 28 were E. baileyi type, 5 were E. cansus type, and the remaining 2 were mixed of E. baileyi + E. cansus (DT2) and E. baileyi + E. smithi (ZN3). The results showed that, the regions around the Qinghai lake and near the upper stream of Yellow River started at Guide could be viewed as the producing area of authentic Sailonggu, and also, the cytb gene is a powerful molecular marker to determine the species of zokors as well as for the authentication of geographic distribution of Sailonggu.
Animals ; Bone and Bones ; metabolism ; Haplotypes ; Medicine, Tibetan Traditional ; Phylogeny ; Rodentia ; classification ; genetics

The study aims to investigate the embryo-fetus development toxicity of the novel PPAR-δ agonist HS060098 on SD rats. The pregnant rats that were randomly divided into the solvent control group (1% hydroxypropyl methyl cellulose water solution) and HS060098 suspension groups (10, 30 and 100 mg x kg(-1) xd(-1)) were orally administered with HS060098 suspension or vehicle during the gestation of 6 -15 days (GD6-15). At termination (GD20), female rats were sacrificed. The pregnant females were evaluated by corpora lutea count, implantation sites, existence and death of embryos. Fetal sex, weight, externals, variations and malformations of viscus and skeleton were observed. The results show that there were no significant abnormality in maternal general conditions and fetal appearance as well as viscera, but in the 100 mg x kg(-1) x d(-1) group, the maternal weight gain decreased greatly (P < 0.01) and the skeletal ossification delayed remarkably (P < 0.01); in the 30 mg x kg(-1) xd(-1) group, the fatal and litter number of incompletely ossified sternebrae II was higher than those of the control group (P < 0.05); the skeletal malformations occurred in all dose groups, which indicate that the novel PPAR-δ agonist HS060098 had maternal toxicity and adversely effected fetal skeletal development under the experimental conditions.
Animals ; Bone and Bones ; drug effects ; Embryonic Development ; drug effects ; Female ; Fetal Weight ; PPAR delta ; agonists ; Pregnancy ; Rats ; Toxicity Tests

Alanine Transaminase ; metabolism ; Animals ; Liver ; drug effects ; Male ; Mice ; Plant Extracts ; pharmacology

Carotid Artery Injuries ; therapy ; Carotid-Cavernous Sinus Fistula ; therapy ; Embolization, Therapeutic ; methods ; Humans ; Male ; Middle Aged ; Stents

PURPOSE: Interleukin (IL)-13, a Th2-type cytokine, plays a pivotal role in the pathogenesis of asthma through its direct effects on airway smooth muscles. A naturally occurring IL-13 polymorphism, R110Q, is strongly associated with increased total serum IgE levels and asthma. In the present study, we aimed to determine whether the IL-13 R110Q variant would display different biochemical properties or altered functions in comparison with wild-type (WT) IL-13 in cultured human bronchial smooth muscle cells (hBSMCs). METHODS: Culture supernatants and cell proteins were collected from cultured hBSMCs that were treated with 50 ng/mL IL-13 or IL-13 R110Q for 24 hours. Eotaxin released into hBSMC culture medium was determined by ELISA. The expression levels of the high-affinity IgE receptor (FcepsilonRI) alpha-chain, smooth muscle-specific actin alpha chain (alpha-SMA), smooth muscle myosin heavy chain (SmMHC), and calreticulin in the cells were measured on Western blots. RESULTS: Compared with WT IL-13, treatment with the IL-13 R110Q variant resulted in a significant increase in eotaxin release as well as significant, although modest, increases in the expression levels of alpha-SMA, SmMHC, calreticulin, and FcepsilonRI alpha-chain. CONCLUSIONS: The results of the present study suggenst that the IL-13 R110Q variant may enhance enhanced functional activities in hBSMCs.
Actins ; Asthma ; Calreticulin ; Enzyme-Linked Immunosorbent Assay ; Humans ; Immunoglobulin E ; Interleukin-13 ; Interleukins ; Muscle, Smooth ; Myocytes, Smooth Muscle ; Myosin Heavy Chains ; Proteins

PURPOSE: Interleukin (IL)-13, a Th2-type cytokine, plays a pivotal role in the pathogenesis of asthma through its direct effects on airway smooth muscles. A naturally occurring IL-13 polymorphism, R110Q, is strongly associated with increased total serum IgE levels and asthma. In the present study, we aimed to determine whether the IL-13 R110Q variant would display different biochemical properties or altered functions in comparison with wild-type (WT) IL-13 in cultured human bronchial smooth muscle cells (hBSMCs). METHODS: Culture supernatants and cell proteins were collected from cultured hBSMCs that were treated with 50 ng/mL IL-13 or IL-13 R110Q for 24 hours. Eotaxin released into hBSMC culture medium was determined by ELISA. The expression levels of the high-affinity IgE receptor (FcepsilonRI) alpha-chain, smooth muscle-specific actin alpha chain (alpha-SMA), smooth muscle myosin heavy chain (SmMHC), and calreticulin in the cells were measured on Western blots. RESULTS: Compared with WT IL-13, treatment with the IL-13 R110Q variant resulted in a significant increase in eotaxin release as well as significant, although modest, increases in the expression levels of alpha-SMA, SmMHC, calreticulin, and FcepsilonRI alpha-chain. CONCLUSIONS: The results of the present study suggenst that the IL-13 R110Q variant may enhance enhanced functional activities in hBSMCs.
Actins ; Asthma ; Calreticulin ; Enzyme-Linked Immunosorbent Assay ; Humans ; Immunoglobulin E ; Interleukin-13 ; Interleukins ; Muscle, Smooth ; Myocytes, Smooth Muscle ; Myosin Heavy Chains ; Proteins

To explore the method of explants directly induced bud and establish the tissue culture system of mutiple shoot by means of direct organogenesis, core bud and daughter bulbs (the top of bud stem expanded to form daughter bulb) of T. edulis were used as explants and treated with thidiazuron (TDZ) and 1-naphthlcetic acid (NAA). The results showed that the optimal medium for bud inducted form core bud and daughter bulb were MS + TDZ 2.0 mg x L(-1) + NAA 4.0 mg x L(-1) and MS +TDZ 2.0 mg x L(-1) + NAA 2.0 mg x L(-1) respectively, both of them had a bud induction rate of 72.92%, 79.22%. The optimal medium for cluster buds multiplication was MS + TDZ 0.2 mg x L(-1) + NAA 0.2 mg x L(-1), and proliferation coefficient was 2.23. After proliferation, cluster buds rooting occurred on MS medium with IBA 1.0 mg x L(-1) and the rooting rate was 52.6%, three to five seedlings in each plant. Using core bud and daughter bulb of T. edulis, the optimum medium for adventitious bud directly inducted from daughter bulb, core bud and cluster bud multiplication were screened out and the tissue culture system of multiple shoot by means of direct organogenesis was established.
Naphthaleneacetic Acids ; pharmacology ; Phenylurea Compounds ; pharmacology ; Plant Growth Regulators ; pharmacology ; Plant Shoots ; drug effects ; growth & development ; Plant Stems ; drug effects ; growth & development ; Seedlings ; drug effects ; growth & development ; Thiadiazoles ; pharmacology ; Tissue Culture Techniques ; Tulipa ; drug effects ; growth & development

BACKGROUNDMuscles present different responses to muscle relaxants, a mechanism of importance in surgeries requiring facial nerve evoked electromyography under general anaesthesia. The non-depolarizing muscle relaxants have multiple reaction formats in the neuromuscular junction, in which pre-synaptic quantal release of acetylcholine was one of the important mechanisms. This study was to compare the pre-synaptic quantal release of acetylcholine from the neuromuscular junctions innervated by normal/damaged facial nerves and somatic nerve under the effect of rocuronium in rats in vitro.

METHODSAcute right-sided facial nerve injury was induced by nerve crush axotomies. Both sided facial nerve connected orbicularis oris strips and tibial nerve connected gastrocnemius strips were isolated to measure endplate potentials (EPP) and miniature endplate potentials (MEPP) using an intracellular microelectrode gauge under different rocuronium concentrations. Then, the pre-synaptic quantal releases of acetylcholine were calculated by the ratios of the EPPs and the MEPPs, and compared among the damaged or normal facial nerve innervated orbicularis oris and tibial nerve innervated gastrocnemius.

RESULTSThe EPP/MEPP ratios of the three neuromuscular junctions decreased in a dose dependent manner with the increase of the rocuronium concentration. With the concentrations of rocuronium being 5 µg/ml, 7.5 µg/ml and 10 µg/ml, the decrease of the EPP/MEPP ratio in the damaged facial nerve group was greater than that in the normal facial nerve group. The decrease in the somatic nerve group was the biggest, with significant differences.

CONCLUSIONSRocuronium presented different levels of inhibition on the pre-synaptic quantal release of acetylcholine in the three groups of neuromuscular junctions. The levels of the inhibition showed the following sequence: somatic nerve > damaged facial nerve > normal facial nerve. The difference may be one of the reasons causing the different sensitivities to rocuronium among the muscles innervated by the normal/injured facial nerves and the somatic nerve. The results may provide some information for the proper usage of muscle relaxants in surgeries requiring electromyographic monitoring for the pre-surgically impaired facial nerves.

Acetylcholine ; metabolism ; Androstanols ; pharmacology ; Animals ; Facial Nerve ; drug effects ; metabolism ; Male ; Neuromuscular Junction ; drug effects ; metabolism ; Rats ; Rats, Sprague-Dawley

Female ; Glomus Tumor ; diagnostic imaging ; pathology ; Humans ; Middle Aged ; Stomach Neoplasms ; diagnostic imaging ; pathology ; Tomography, Spiral Computed ; methods