Objective: To investigate a new strategy to reverse multidrug resistance of chronic myeloid leukemia cell line K562 by RNA interference technique through constructing an eukaryotic vector of short hairpin RNA (shRNA) targeting homeobox A10 (HOXA10) gene. Methods: The eukaryotic vector pGPHI/GFP/Neo-HOXA10 with shRNA targeting HOXA10 gene was constructed and then transfected into K562 cells by positive ion liposome. The stable transfectants were vertificated by reverse transcriptase PCR (RT-PCR) 4 weeks after G418 pressure selection. The changes of sensitivity of K562 cells to leurocristine (VCR) and etoposide (VP-16) after transfection with shRNA-HOXA10 were detected by MTT method, and the apoptosis rate was detected by flow cytometry. Results: After selection with G418, the cell clones stably transfected with pGPHI/GFP/Neo-HOXA10 were successfully constructed and verificated by RTPCR. The half inhibitory concentration (IC50) values of VCR and VP-16 for K562 cells transfected with shRNA-HOXA10 were significantly reduced and the apoptosis rate of K562 cells after HOXA10 gene interference combining with chemotherapy was increased significantly as compared with those of shRNA-negative control group and the normal control group (P<0.05). No differences in IC50 and apoptosis rate were observed between the shRNA-negative control group and the normal control group (P>0.05). Conclusion: The eukaryotic vector of short hairpin RNA (shRNA) targeting HOXA10 gene can enhance the abilities of VCR and VP-16 to inhibit the cell proliferation and induce the apoptosis of K562 cells, namely an increase of sensitivity to these chemotherapeutics in K562 cells. It is hinted that RNA interference targeting HOXA10 gene may reverse the multidrug resistance of leukemia cells in some degree. Copyright© 2011 by TUMOR.

Objective To observe the effect of pulmonary surfactant(PS) on lung function and ventilator parameters of neonatal respiratory distress syndrome(NRDS) in the advanced stage.Methods Twenty-eight infants with NRDS were given PS in one dose by endotracheal intubation on the left side,right side,feet high with head low,and level decub respectively.The dose of PS was 100-150 mg/kg each time,each posture slow note of the drug were required 1/4,out of the straw,hand-controlled ventilation,to reduce fluid loss,with the exception of a clear airway obstruction,within 6 hours after the administration not to shoot back suction,to give mechanical ventilation after the injection.Lung function parameters were also measured:pressure of oxygen in artery[p_a(O_2)],carbon dioxide partial pressure[p_a(CO_2)],the ratio of pressure of oxygen in artery and alveolar oxygen partial pressure[a/Ap(O_2)] and oxygenation index(OI) were determined.Ventilator parameters were determined:oxygen concentration(FiO_2),oxygen peak(PIP),end-expiratory positive pressure(PEEP) and mean airway pressure(MAP) were determined.These numerical data were analyzed and compared before and after treatment with PS.Clinical manifestations,thoracic X-ray changes,survival rate and incidence rate of complications were also analyzed and compared before and after PS therapy.Results p_a(O_2),a/Ap(O_2) showed significant upgrade and OI had a decrease after PS administration in comparison with those before PS therapy.The ventilator parameters(except for PEEP) acquired were also lower after drug administration than those in before drug therapy.There were significant differences in both stages(P_a90%,respiratory sound in 24 cases enhanced,the observation of chest film after 24 h indicated that,lesions in 21 cases improved significantly,5 cases took a favorable turn.The survival rate was 85.7%.The incidence rate of complication was as follows:pneumonia was 25%,patent ductus arteriosus was 10.7%,pneumorrhagia was 7.1% and intraventricular hemorrhage was 3.6%,respectively.Conclusion Respiratory function of NRDS is significantly improved by using PS in the advanced stage,and therapeutic effect is apparent.

This paper aims to establish a method for the determination of sulfur dioxide in sulfur fumigation Chinese herbs. Sample powder and hydrochloric acid solution were isolated by paraffin layer in order to avoid early reactions, with the generation of sulfur dioxide, headspace with airtight needle was used to transfer sulfur dioxide into gas chromatograph, and detected with thermal conductivity detector. The analytical performance was demonstrated by the analysis of 12 herbs, spiked at four concentration levels. In general, the recoveries ranging from 70% to 110%, with relative standard deviations (RSDs) within 15%, were obtained. The limit of detection (LOD) was below 10 mg x kg(-1). Standard addition can be used for low recovery samples. The method is simple, less time-consuming, specific and sensitive. Methods comparison revealed that gas chromatography is better than traditional titration in terms of method operability, accuracy and specificity, showing good application value.
Chromatography, Gas ; methods ; Fumigation ; Limit of Detection ; Plants, Medicinal ; chemistry ; Sulfur ; chemistry ; Sulfur Dioxide ; analysis

The content of the asarone submicro emulsion injection was determind by HPLC method, and thereby a quality evaluation method was established based on indexes of pH value, particle size, peroxide value, methoxy aniline values, free fatty acid, lysophosphatidylcholine, visible foreign substances, insoluble particle, sterility, bacterial endotoxin and impurities, etc. The results showed that the injection exhibited uniform physical appearance and all the products were in milkwhite liquid. The content of the three batches products were respectively 102.9%, 100.8%, 97.70% of the labeled amount, with mean particle size of 210-250 nm, and other indexes all met with the standards. The reserved samples showed no obvious change in terms of detection indexes and indicated good stability after the accelerated stability test and long-term stability for 12 months. The quality evaluation method established in this study could be applied to quality control and stability investigation of asarone submicron emulsion injection, which laid a basis for further clinical research and application.
Anisoles ; chemistry ; Chromatography, High Pressure Liquid ; Drug Stability ; Drugs, Chinese Herbal ; chemistry ; Emulsions ; chemistry ; Particle Size ; Quality Control

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Carcinoma, Hepatocellular ; diagnosis ; epidemiology ; virology ; Child ; China ; epidemiology ; Female ; Genes, Viral ; Genotype ; Hepatitis B virus ; genetics ; Humans ; Liver Neoplasms ; diagnosis ; epidemiology ; virology ; Male ; Middle Aged ; Viral Load ; Young Adult

OBJECTIVETo study the dynamic changes in the protein marker expression in the spermatogonial stem cells (SSCs) of mice at different ages by iTRAQ protein mass spectrometry and to screen new markers using the bioinformatic proteome database.

METHODSBased on the postnatal weeks, we divided 80 healthy male C57BL/6 mice into eight age groups of equal number, harvested their testicular tissues, extracted proteins following purification of the SSCs by compound enzyme digestion and magnetic-activated cell sorting. Then we analyzed and identified proteins using two-dimensional electrophoresis, protein mass spectrometry, and protein database information.

RESULTSTotally, 248,510 mass spectra were obtained from the MS experiment and 1132 proteins were identified. By the criteria of >1.2-fold for protein abundance difference and P value <0.05, we identified 298 differentially expressed proteins and 9 currently known makers of SSCs (PCNA, GFRalpha1, CDH1, Annexin A7, UCHL1, VASA, CD49f, CD29, and PLZf). Compara- tive analysis showed different expressions of the proteins in the SSCs of the mice of different ages, and the differences in the expressions of GFRalpha1, CD49f, and CD29 were consistent with the findings in other published literature. Ten proteins (P63, CD71, CD98, K19, ACE, K18, K15, K17, SH2, and SH3) were selected as SSC markers to be further studied.

CONCLUSIONThe proteins in SSCs are differentially expressed in mice of different ages. The technology of iTRAQ protein mass spectrometry can be used to analyze and compare the proteome information of mouse SSCs, obtain differentially expressed proteins in mice of different ages, and thus offers a new ap- proach to further analysis and study of the function and roles of these differential proteins.

Adult Stem Cells ; cytology ; metabolism ; Age Factors ; Animals ; Biomarkers ; analysis ; metabolism ; Cell Separation ; methods ; Electrophoresis, Gel, Two-Dimensional ; Male ; Mass Spectrometry ; Mice ; Mice, Inbred C57BL ; Proteins ; analysis ; metabolism ; Spermatogonia ; cytology

Hydrolytic amino acids were extracted by acid hydrolysis method, then derivatized with phenyl isothiocyanate (PITC). And the samples were analysed by HPLC on an Ultimate Prime C18 (4.6 mm x 250 mm, 5 microm) column with gradient elution of 0.1 mol x L(-1) sodium acetate buffer solution (adjusted to pH 6. 5)-acetonitrile (93:7) (A) and acetonitrile-water (8:2) (B) at a flow rate of 1.0 mL x min(-1). Column temperature was 40 degrees C and the detected wavelength was 254 nm. Amino acids derivative solution remained stable in 36 hours. The response was linear for 16 amino acids with a correlation coefficient r > 0.999 5. The average recoveries were 98.01% -101.8%. The method is reliable with good accuracy and repeatability, which is useful for the determination of amino acids in Galli Gigerii Endothelium Corneum.
Amino Acids ; analysis ; Animals ; Chickens ; Chromatography, High Pressure Liquid ; Chromatography, Reverse-Phase ; Endothelium ; chemistry ; Gizzard, Avian ; chemistry

AIM: To report a two- year's results of iontophoresis-assisted transepithelial corneal cross-linking (I-CXL) for progressive keratoconus.
METHODS: Thirty - four eyes in 24 patients with progressive keratoconus ( mean age 21. 0 ± 5. 6 years;range: 14-32 years) were treated. After 1g/ L riboflavin-distilled water solution was administered by iontophoresis-assited (current 1mA) transepithelial method for 5min in total, standard surface UVA irradiation ( 370nm, 3mW/cm2 ) was performed at a 1 - cm distance for 30min. The best corrected visual acuity ( BCVA ) measured as LogMAR number, corneal refractive astigmatism, K1, K2, Kmean, Kmax, intraocular pressure, endothelial cell density, the thickness at corneal apex and the thinnest point were measured preoperatively and 2a postoperatively.
RESULTS:At 2a after the procedure, BCVA (LogMAR) improved from 0. 32 ± 0. 25 to 0. 25 ± 0. 19 ( t = 2. 849, P =0. 015). K1 decreased from 47. 12±4. 33 to 46. 06±4. 77 (t =2. 652, P= 0. 015). K2 decreased from 51. 36±5. 59 to 50. 40±6. 16 (t= 2. 121, P= 0. 047). Kmean decreased from 49. 12±4. 76 to 48. 10±5. 25(t = 2. 663, P = 0. 015). Kmax decreased from 57. 57±8. 30 to 55. 91±8. 14 (t = 2. 398, P = 0. 026). The corneal apex thickness decreased from 476. 90±38. 71μ m to 454. 43 ± 40. 86μ m ( t = 2. 853, P = 0. 010 ). The thinnest thickness decreased from 464. 38 ± 39. 92μ m to 433. 86 ±50. 78μ m ( t = 3. 485, P = 0. 002 ). Corneal refractive astigmatism, intraocular pressure and endothelial cell density did not show significant changes.
CONCLUSION: I - CXL for progressive keratoconus is safe and effective which can prevent deterioration of progressive keratoconus within 2a, but further long-term studies are necessary still.

An antibiotic-producing bacterium, which was numbered as 20 #-5, was separated from the soil in Chongqing. It was identified as the member of pseudomonas. Gram positive bacteria are badly suppressed by it. The antibiotic secreted by 20 #-5 can endure 100℃ for half an hour, and it can also go through the ultrafiltration membrane with pores of 0.22?m.

Objective To evaluate the clinical value of color doppler flow image (CDFI) and three-dimension color power angiography(3D-CPA) in the diagnosis of thyroid nodules. Methods A total of 62 pathologically confirmed thyroid nodules of 30 patients were analyzed. All the patients underwent ultrasound examination and operation in the Second Affiliated Hospital of Harbin Medical University between January, 2008 and May, 2009. Both CDFI and 3D-CPA were performed before the operation. All the nodules were divided into three groups including nodular goiters, thyroid adenomas and thyroid cancers according to the pathological results. The hemodynamic features and the vascular morphology characteristics of nodules in different groups were compared. Results 3D-CPA showed that blood vessels of nodular goiters commonly distributed surrounding the mass and the inner vessels were thin and regular, thyroid adenomas were bulb-shaped and netted structure, and malignant thyroid nodules displayed distorted and irregular distributed vessels. Peak systolic velocity (PSV)of the three groups were (39.43±11.17a), (46.39±12.98) and (65.17±9.23)cm/s, respectively. Resistance index(RI) of the three groups were (0.32±0.08), (0.41±0.06) and (0.69±0.07)cm/s, respectively. Both PSV and RI in malignant thyroid nodules were higher than in nodular goiters and in thyroid adenomas and the difference were statistical significant (all P < 0.05). The blood flow grade of malignant nodules was also higher in malignant nodules than in other two groups(χ2 = 17.11, 12.79, 23.05, 15.41, P< 0.01). Conclusions CDFI and 3D-CPA could visually demonstrate the characteristic and distribution of the inner and outer blood vessels, display the vessels structures, and they are benefit the differential diagnosis of thyroid nodules.