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.

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

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

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

OBJECTIVETo study the characteristics of the SiO2/R2O3 in non-zonal purple soil in the light of the difference between the high yield and low yield of Salvia miltiorrhiza.

METHODBoth the high yield and low yield drugs developed from the same parental material (k1cy) were chosen in the ecology area in Zhongjiang county, Sichuan Province. By using the air-dried drugs as sample, quadrat sampling method was used to calculat the production. Gravimetric method, volumetric method, and colorimetric method and neutralization titration were used to measure the content of adhesive particle colloids element in the soil. And T-test was used for the variable-difference analysis.

RESULTThere was a sharp contrast between the production of high yield and low yield drugs (P < 0.001). The characteristic value of the high yield soil embryology was much higher than that of the low yield soil.

CONCLUSIONThe characteristic value of the purple soil embryology is one of the important factors of the soil adaptability of the drug and its quality. The weathering degree of the soil is closely related to the yield of the drug.

Aluminum ; analysis ; Aluminum Oxide ; analysis ; Ecosystem ; Ferric Compounds ; analysis ; Iron ; analysis ; Plants, Medicinal ; growth & development ; Salvia miltiorrhiza ; growth & development ; Silicon ; analysis ; Silicon Dioxide ; analysis ; Soil ; analysis ; Titanium ; analysis

OBJECTIVETo assess the nasal airway changes after maxillary advancement following Le Fort I osteotomy.

METHODS13 cases with class III malocclusion, aged 18-35 years old, were studied prospectively. All the patients underwent Le Fort I osteotomy and maxillary advancement. Rhinological inspectrum, acoustic rhinometry (AR) were performed before operation, 3 and 6 months after operation. The Nasal Obstruction Symptom Evaluation (NOSE) scale was also completed by 13 patients before and after operation. SPSS was used for statistical assay.

RESULTSAR assessment showed that NAR was (1.189 +/- 0.38) cm H2O/L/mi, (1.081 +/- 0.43) cm H2O/L/mi and (1.111 +/- 0.40) cm H2O/L/mi before operation, 3 and 6 months after operation; NV was (14.920 +/- 1.95) ml, (16.380 +/- 4.32) ml and (15.660 +/- 4.25) ml; and MCA was (0.500 +/- 0.09) cm2, (0.570 +/- 0.15) cm2 and (0.560 +/- 0.14) cm2, respectively. However, no significant improvement was showed. For the whole cohort, significant improvement in nasal breathing was documented (by NOSE scores) at 6 months after surgery.

CONCLUSIONSLe Fort I osteotomy with maxillary advancement doesn't cause bad effect on nasal airways in patients with maxillary dysplasia. And the combination of objective (AR) and subjective (NOSE scale) assessment can better evaluate of the structure and function of the nose.

Adolescent ; Adult ; Female ; Humans ; Male ; Maxilla ; surgery ; Nose ; physiopathology ; Osteotomy, Le Fort ; methods ; Postoperative Period ; Respiration ; Young Adult

OBJECTIVETo investigate serum procalcitonin (PCT) concentrations in premature infants with different gestational ages at different times after birth.

METHODSA total of 217 neonates without infection, including 102 premature infants and 115 full-term infants, were enrolled in this study. The premature infants were further divided by gestational age into three subgroups: 30-32 weeks (n=30), 33-34 weeks (n=35) and 35-36 weeks (n=37). All the infants were studied to evaluate serum PCT concentrations at 0-12, 13-24, 25-36, 37-48, 49-72, 73-96, 97-120 and 121-144 hours after birth.

RESULTSIn the newborns, serum PCT concentrations increased gradually after birth, reached peak values at about 24 hours after birth, and then gradually declined and dropped to normal values for children at about 96 hours after birth. In the premature infants, serum PCT concentrations reached peak values at about 36 hours after birth, later than in the full-term infants, then declined slowly and dropped to levels similar to the full-term infants at 96 hours after birth. Serum PCT concentrations in the 30-32 week subgroup remained at low levels after birth, and increased gradually, later than in other premature infants, at 37-48 hours after birth.

CONCLUSIONSEarly after birth, neonates have a changing serum PCT concentration, increasing first and then decreasing. Peak serum PCT levels appear later in premature infants than in full-term infants. Serum PCT concentrations of premature infants with a gestational age of under 32 weeks remain at relatively low levels within 36 hours after birth.

Calcitonin ; blood ; Calcitonin Gene-Related Peptide ; Gestational Age ; Humans ; Infant, Newborn ; Infant, Premature ; blood ; Protein Precursors ; blood ; Time Factors