Adolescent ; Adult ; Combined Modality Therapy ; Female ; Fracture Fixation ; methods ; Humans ; Male ; Manipulation, Orthopedic ; methods ; Middle Aged ; Splints ; Ulna Fractures ; therapy

OBJECTIVETo construct a chimeric SEA-hPLAP-1 cDNA with gene splicing by overlap extension.

METHODSThe SEA gene and a DNA fragment encoding the signal for GPI-anchor attachment of hPLAP -1 were amplified by PCR. The two amplified gene sequence was annealed to form a chimeric GPI- anchored SEA molecule with gene splicing by overlap extension. The resulting chimera was cloned in pGEM-T vector and verified by sequencing analysis.

RESULTA chimeric SEA-hPLAP-1 cDNA was successfully constructed with gene splicing by overlap extension.

CONCLUSIONGene splicing by overlap extension is a successful specific PCR technique for gene recombination.

Alkaline Phosphatase ; Base Sequence ; Enterotoxins ; genetics ; GPI-Linked Proteins ; Isoenzymes ; genetics ; Molecular Sequence Data ; Polymerase Chain Reaction ; RNA Splicing ; Recombinant Fusion Proteins ; genetics

OBJECTIVETo investigate the potential role of caveolin-1 (CAV-1) on membrane estrogen receptor (mER) mediated proliferation of endothelial progenitor cells (EPCs).

METHODSBone marrow (BM)-derived EPCs were cultured. The proliferation of EPCs induced by estradiol (E₂)-BSA in the absence or presence of ICI 182, 780 (a pure ER inhibitor), MβCD and CAV-1 siRNA was determined by [³H]-thymidine incorporation. The expression of CAV-1 was detected by Western blot.

RESULTSProliferation of EPC peaked after 10(-8) mol/L E₂-BSA culture for 24 h (87.5% increase vs. control), and this effect could be inhibited by estrogen receptor blocker ICI 182, 780, indicating that mER-initiated membrane signaling pathways was involved in the proliferation effect of estrogen on EPC. Both cholesterol depletion and CAV-1 siRNA significantly attenuated E₂-BSA induced [³H]-thymidine incorporation. Western blot result confirmed that cholesterol depletion or CAV-1 siRNA significantly decreased CAV-1 protein expression (-18.6% or -41.2% vs. 10(-8) mol/L E₂-BSA alone).

CONCLUSIONOur results suggested that estradiol promoted EPC proliferation through activating CAV-1 pathway.

Animals ; Caveolin 1 ; immunology ; pharmacology ; Cell Proliferation ; drug effects ; Cells, Cultured ; Endothelial Cells ; cytology ; metabolism ; Endothelium, Vascular ; cytology ; metabolism ; Estradiol ; metabolism ; Rats ; Rats, Sprague-Dawley ; Receptors, Estrogen ; metabolism ; Stem Cells ; cytology ; metabolism

OBJECTIVETo explore the inhibition and molecular mechanism of icaritin (ICT) combined doxorubicin (DOX) on human osteosarcoma MG-63 cells in vitro.

METHODSThe control group, ICT groups (10, 20, 40, 80, and 160 µmol/L), DOX groups (1, 2, 4, 8, and 16 µg/mL), and combination groups (20 µmol/ L ICT +1 µg/mL DOX, 20 µmol/L ICT +2 µg/mL DOX, 20 µmol/L ICT +4 µg/mL DOX, 40 µmol/L ICT +1 µg/mL DOX, 40 µmol/L ICT +2 µg/mL DOX, 40 µmol/L ICT +4 µg/mL DOX, 80 µmol/L ICT +1 µg/mL DOX, 80 µmol/L ICT +2 µg/mL DOX, 80 µmol/L ICT +4 µg/mL DOX) were set up. Human osteosarcoma MG-63 cells were respectively cultured and their effects on morphological changes were observed using inverted phase contrast microscope after 24-and 48-h intervention. The cell proliferation inhibition rate of each group was de- termined using CCK-8, and IC50 calculated. The MG-63 apoptosis rate was detected using Annexin V-FITC/ PI double dye flow cytometry. Expression levels of bcl-2, caspase-3, and p21 were detected using RT-PCR.

RESULTSICT and DOX could obviously inhibit the proliferation of MG-63 cell. Along with ICT concentration increasing from 10 µmol/L to 160 µmol/L, the cell proliferation inhibition rate also increased gradually from 9.67% ± 3.62% to 89.18% ± 9.66%. The IC50 was 46.93 µmol/L and 3.87 µg/mL respectively. ICT and DOX could cause either early or late stage apoptosis, down-regulate Bcl-2 gene expression, and up-regulate gene expressions of Caspase-3 and p21 respectively (P < 0.05). Aforesaid changes were more obviously seen in combination groups than in lCT groups and DOX groups (P < 0.05).

CONCLUSIONCT combined DOX had additive or synergistic inhibition effect for the proliferation of osteosarcoma MG-63 cells, which might be related with regulating gene expressions of bcl-2, caspase-3, and p21.

Apoptosis ; Bone Neoplasms ; metabolism ; pathology ; Caspase 3 ; metabolism ; Cell Line, Tumor ; drug effects ; Cell Proliferation ; Cyclin-Dependent Kinase Inhibitor p21 ; metabolism ; Down-Regulation ; Doxorubicin ; pharmacology ; Drug Synergism ; Flavonoids ; pharmacology ; Humans ; Osteosarcoma ; metabolism ; pathology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism

The study found that the presence of intestinal microbiota is not only important for the metabolism of essential nutrients in the body, but also plays a key role in the development of the body′s immune system in recent years. Partial microbiota, through natural selection and co-evolution with the host, forms symbiotic relationships with host microbes that are inseparable from host physiology in mice. Symbiotic flora affects the formation of the body′s immune system by affecting innate and adaptive immunity and the development of various regulatory mechanisms. The destruction of the microbial ecosystem in the intestine can lead to the occurrence of many diseases,especially those related to the immune system. Peripheral immune organs always receive a number of immune cells colonized by antigen stimulation. So,the intestinal flora plays an important role in maintaining the function of immune cells. This article will investigates the effects of mouse-related intestinal flora on peripheral immune organ function.

Long-term excessive iodine intake resulted in an increased TT_4 level and a decreased TT_3 level in maternal serum,meanwhile,hepatic and renal type 1 deiodinase activity decreased dose-dependently.A significant reduction in type 2 deiodinase ( D2 ) activity of 12.5 d placenta was found in 3.0 mg/L or above groups.For 19.5 d uterus,D2 activity decreased and type 3 deiodinase activity increased.The results suggest that excessive iodine has an effect on the embryonic development by regulating maternal-fetal thyroid hormone metabolism.

OBJECTIVETo establish and optimize the two-dimensional electrophoresis (2-DE) of uterine leiomyoma for the proteome analysis.

METHODSRun immobilized pH gradient (IPG)-isoelectric focusing electrophoresis as the first dimension, then vertical SDS-PAGE electrophoresis as the second dimension. A series of important steps,such as sample solubility, volume of loading, electrophoresis parameters and protocol for staining were optimized.

RESULTSThe 2-DE patterns of uterine leiomyoma and myometrium with good quality were obtained.

CONCLUSIONWith optimal condition the two-dimensional electrophoresis of uterine leiomyoma can be obtained.

Electrophoresis, Gel, Two-Dimensional ; Female ; Humans ; Leiomyoma ; chemistry ; Myometrium ; chemistry ; Neoplasm Proteins ; analysis ; Proteome ; analysis ; Uterine Neoplasms ; chemistry

OBJECTIVE: To explore the feasibility of detecting of Y-STR of fetal DNA in maternal plasma using Ion Torrent PGM™ System. METHODS: A total of 16 fetal DNA samples from maternal plasmas (8 cases from 38 weeks gestational age and 8 ones from 12 weeks) were prepared and a multiplex assay with 7 STR loci (DYS390, DYS391, DYS393, DYS438, DYS437, DYS456, DYS635) was designed for multiplex-PCR amplification. Using Ion Torrent PGM™ System, the results of Y-STR sequences and capillary electrophoresis were obtained and compared. RESULTS: Y-STR specific alleles were detected in the maternal plasma of all the pregnant women having male babies of second and third trimester, which were higher than that detected by capillary electrophoresis. Consistent Y-STR genotypes were observed between fetal DNA from maternal plasma and genomic DNA from the newborn babies. CONCLUSION Based on Ion Torrent PGM™ System, the prenatal Y-STR detection method may provide a high-sensitive and high-throughput choice for prenatal STR detection in forensic testing.
Alleles ; Chromosomes, Human, Y/genetics* ; DNA/blood* ; Family ; Female ; Fetal Blood/chemistry* ; Genotype ; Haplotypes ; Humans ; Male ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Pregnancy ; Sensitivity and Specificity ; Sex Determination Analysis ; Tandem Repeat Sequences/genetics*

OBJECTIVETo investigate the appropriate extubation time and treatment of late complications after early tracheotomy in patients with moderate or severe inhalation injury.

METHODSOne hundred and fifty patients (105 males and 45 females) with inhalation injury were admitted to our hospital from January 2000 to January 2009. Among them, 109 out of 129 cases with moderate inhalation injury received early tracheotomy, and all 21 cases with severe inhalation injury received early tracheotomy. Data were collected for analysis as follows: (1) incidence of re-intubation due to suffocation and pneumonia incidence after extubation within 2 weeks or after 2 weeks post inhalation injury (PII), and mortality rate within the first week after injury were recorded. (2) Conservative treatments including expectorant, oral antibiotics, and absolute bedrest were recommended for patients who had severe cough, hoarseness or poor pulmonary function after late extubation and closure of tracheostomy wound. Fiberoptic bronchoscopy findings (tracheostenosis degree, granuloma formation rate, vocal cord paralysis rate) and pulmonary function index (FEV(1)) data were collected and analyzed in 30 cases with moderate inhalation injury and 10 cases with severe inhalation injury within 3 months after injury for follow-up. Data were processed with t test or chi-square test.

RESULTSThere was no obvious difference in the rate of re-intubation after extubation in patients with moderate inhalation injury between those done within 2 weeks PII (15/70, 21.4%) and those done after 2 weeks PII (2/25, 8.0%) (χ(2) = 1.52, P > 0.05). Pneumonia incidence in patients of moderate inhalation injury with extubation within 2 weeks PII (21/70, 30.0%) was lower than those with extubation after 2 weeks PII (15/25, 60.0%) (χ(2) = 7.04, P < 0.05). Levels of above-mentioned indexes in patients with severe inhalation injury extubated in different stages were similar to those of patients with moderate inhalation injury. Within the first week after injury, mortality rate of patients with severe inhalation injury was higher than that of patients with moderate inhalation injury (χ(2) = 11.90, P < 0.05). During follow-up, tracheostenosis rate in patients with moderate or severe inhalation injury was 100.0%; granuloma formation rate and vocal cord paralysis rate in patients with severe inhalation injury were higher than those of patients with moderate inhalation injury (with χ(2) value respectively 4.59, 13.47, P values all below 0.05). The FEV(1) value of patients with moderate inhalation injury in the 1st, 2nd, 3rd month after injury was respectively higher than that of patients with severe inhalation injury (with t value respectively 5.48, 12.10, 6.25, P values all below 0.05). The values recovered to normal level in the 3rd month after injury.

CONCLUSIONSExtubation time of tracheotomy for patients with moderate or severe inhalation injury within 2 weeks or after 2 weeks PII has its own advantage and disadvantage, and it should be performed according to specific conditions of each patient. Conservative treatment is optional for late complications of respiratory system.

Adolescent ; Adult ; Aged ; Burns, Inhalation ; surgery ; Child ; Child, Preschool ; Female ; Follow-Up Studies ; Humans ; Intubation, Intratracheal ; adverse effects ; Male ; Middle Aged ; Postoperative Complications ; Tracheotomy ; adverse effects ; Young Adult

AIMTo develop a sensitive and specific HPLC method for determination of trigonelline in rabbit plasma, and study the pharmacokinetics in rabbit.

METHODSAfter ig of fenugreek extract and i.v. of trigonelline in rabbit, the biological samples could be well purified after precipitation of protein with methanol and acetonitrile. Asahipak NH2P-50 column was used, the mobile phase consisted of acetonitrile-water (90:10) at a flow-rate of 1.2 mL.min-1, and detection wavelength was set at UV 265 nm. The column temperature is 30 degrees C.

RESULTSThe calibration curve was linear in the range from 0.98 mg.L-1 to 31.28 mg.L-1, with r = 0.9986, the detection limit of this method was 50 micrograms.L-1. The concentration-time curves of trigonelline in rabbits after ig and i.v. administration were shown to fit one-compartment and two-compartment open model, respectively. The main parameters after ig of fenugreek extract were as follow: T1/2(Ka) was 0.9 h, T1/2(Ke) was 2.2 h, V was 0.64 L.kg-1, AUC was 1.93 mg.min.L-1. The main parameters after i.v. of trigonelline were as follows: T1/2 alpha was 10.8 min, T1/2 beta was 44.0 min, K21 was 0.044 min-1, K10 was 0.026 min-1, K12 was 0.017 min-1, AUC was 931.0 mg.min.L-1.

CONCLUSIONTrigonelline showed a middle rate of absorption and fast rate of elimination in rabbit. Meanwhile, the method is simple, accurate, with a good reproducibility, and it provide a basic method for the investigation of trigonelline and fenugreek pharmacokinetics.

Alkaloids ; blood ; isolation & purification ; pharmacokinetics ; Animals ; Antineoplastic Agents, Phytogenic ; blood ; isolation & purification ; pharmacokinetics ; Area Under Curve ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; isolation & purification ; pharmacokinetics ; Female ; Male ; Plants, Medicinal ; chemistry ; Rabbits ; Seeds ; chemistry ; Trigonella ; chemistry