A method was established for the simultaneous determination of the total fatty acid esters of chloropropanols in edible oils by gas chromatography-mass spectrometry combined with isotope dilution technology. The samples were hydrolyzed with sodium methylate-methanol, and then purified by diatomite cartridge. After being derivatized with heptafluorobutyrylimidazole ( HFBI ), the target analytes were determined by GC-MS with the deuteriumchloropropanols esters as the internal standards. An excellent linear correlation in the range of 0. 050-2. 000 mg / L was acquired for 3-monochloropropane-1,2-diol (3-MCPD) esters, 2-MCPD esters, dichloropropan-2-ol (1,3-DCP) esters and 2,3-dichloropropan-1-ol (2,3-DCP) esters, with all the correlation coefficients (r) higher than 0. 9995. The limits of detection (LODs) for 3-MCPD esters, 2-MCPD esters, 1,3-DCP esters and 2,3-DCP esters were 0. 015, 0. 015, 0. 030, and 0. 030 mg / kg, respectively, and the limits of quantitation (LOQ) were 0. 050, 0. 050, 0. 100, and 0. 100 mg / kg, respectively. The average spike recoveries of the four kinds of chloropropanols esters in blank extra virgin olive oil matrix were typically in a range of 87. 0% -110. 5% with the relative standard deviations (RSDs) less than 10. 1% . The detection rates of 3-MCPD esters, 2-MCPD esters, 1,3-DCP esters and 2,3-DCP esters in 74 edible oil samples were 94. 6% , 63. 5% , 5. 4% , and 0% , respectively. The contamination levels of 3-MCPD esters, 2-MCPD esters and 1,3-DCP esters were in the range of not detected (ND) to 10. 646 mg / kg, ND to 3. 617 mg / kg and ND to 0. 089 mg / kg, respectively. This method is accurate and rugged for the simultaneous determination of total fatty acid esters of chloropropanols in edible vegetable oils.

Objective To establish a content determination method for paeoniflorin in qisheng capsule. Methods The quantitative analysis of paeoniflorin in qisheng Capsule was carried out by high-performance liquid chromatography ( HPLC) . The chromatographic separation was achieved by using a Kromasil C18 chromatographic column (4. 6 mm×250 mm,5 μm) with a mobile phase consisting of methanol,water (1585) at a flow rate of 1. 0 mL·min-1 and 230 nm detection wavelength. Results The linear range was 2. 5-12. 5 μg·mL-1( r =0. 999 9). The average recovery and RSD of the method were 99. 97%and 0. 94%. Conclusion The method is accurate,specific,reproducible,which can effectively be used in quality control of paeoniflorin in qisheng capsule.

Aim To study the effect of Tryptanthrin(Try) on proliferation and apoptosis of erythroleukemia K562 cells.Methods The cell proliferation effect of Try(1.56～50 mg?L-1) on K562 cells was assessed by MTT assay.The morphologic change was observed by Hoechst 33258 fluore-scent stain.The flow cytometer was used to detect cell apoptosis and cell cycle.Results MTT showed that in the range of 3.12～50 mg?L-1 Try obviously inhibited the proliferation of K562 cells in a dose and time-dependent manner.Typical apoptosis changes were observed in K562 cells treated with Try for 48 h by flourescence inverted microscope.With Annexin V-FITC and PI double staining,folw cytometer result showed that the apoptosis state was obvious in K562 cells treated with 25,50 mg?L-1 Try for 48 h.The cell cycle distribution of K562 was changed.The G0/G1 phase was blocked and the DNA synthesis was inhibited,accompanied with subdiploid apoptotic peak.Conclusion Try has an effect on inhibiting the cell proliferation and inducing the apoptosis of K562.

OBJECTIVETo compare the differences in the therapeutic effect on irritable bowel syndrome (IBS) between acupuncture at Tianshu (ST 25) and Dachangshu (BL 25) and western medication with Trimebutine Maleate.

METHODSForty cases were divided randomly into an acupuncture group and a western medication group, 20 cases in each one. In acupuncture group, acupuncture was applied to Tianshu (ST 25) and Dachangshu (BL 25). Ziwu Daojiu needling technique was adopted, once daily. In western medication group, Trimebutine Maleate capsule was administered, 2 capsules in each time, 3 times per day. The assessment on the therapeutic effect was performed in 4 weeks of treatment in two groups.

RESULTSAs compared with those before treatment, the time of abdominal pain, the frequency of abdominal pain, the morbidity of abnormal stool appearance, the morbidity of defecation abnormality, the morbidity of mucus stool and the score of bloating or abdominal pain on bowel movement were all reduced after treatment in two groups (all P < 0.01). The results in acupuncture group were much more significant than those in western medication group (the total score: 16.70 +/- 2.40 vs 15.70 +/- 3.01, P < 0.01). The total effective rate in acupuncture group was 95.0% (19/20), which was superior to that of 70.0% (14/20) in western medication group (P < 0.05).

CONCLUSIONAcupuncture at Tianshu (ST 25) and Dachangshu (BL 25) may remarkably relieve the clinical symptoms of IBS and its efficacy is superior to that of oral medication with Trimebutine Maleate.

Acupuncture Therapy ; Adolescent ; Adult ; Aged ; Child ; Female ; Humans ; Irritable Bowel Syndrome ; drug therapy ; therapy ; Male ; Middle Aged ; Trimebutine ; therapeutic use ; Young Adult

BACKGROUNDGlycoprotein non-metastatic melanoma protein B (GPNMB) plays an important role in the pathogenesis of inflammatory and malignant diseases. We investigated the expression of GPNMB in benign and malignant skin diseases.

METHODSTissue microarray was performed in the skin tissues of 102 cases including malignant melanoma (MM), squamous cell carcinoma (SCC), basal cell carcinoma (BCC), and benign dermatosis. The expression of GPNMB in the tissues was detected by immunohistochemistry. Twenty cases of normal skin and adjacent neoplastic normal skin tissues were selected as controls.

RESULTSGPNMB was positively stained in skin malignancies (38/50, 76%), which was significantly higher than that in the control and the benign skin tissues (P = 0.001 and < 0.001 respectively). GPNMB was positively stained in MM (13/15, 87%) and SCC (16/20, 80%) (P < 0.001). Significant higher expression of GPNMB was observed in patients aged ≥ 65 years than those less than 65 years (n = 11 and n = 9 respectively, P = 0.027). No significant difference of the expression rates was observed between normal control and BCC; however, stronger intensity was detected in the latter. Negative or weak expression was observed in the controls.

CONCLUSIONOver-expression of GPNMB correlated strongly and might play an important role in the pathogenesis of MM and SCC.

Adolescent ; Adult ; Aged ; Carcinoma, Basal Cell ; metabolism ; Carcinoma, Squamous Cell ; metabolism ; Female ; Humans ; Immunohistochemistry ; Male ; Melanoma ; metabolism ; Membrane Glycoproteins ; metabolism ; Middle Aged ; Skin ; metabolism ; pathology ; Skin Diseases ; metabolism ; Skin Neoplasms ; metabolism ; Tissue Array Analysis ; methods ; Young Adult

Objective To develop a method of quantitative analysis of multi-components by single marker (QAMS) for simultaneously determining five compounds in Curcumae Aromaticae Radix.Methods An HPLC method was developed as QAMS to determine curcuma diol,ocathydro-1,4-dihydroxy-1,4-dimethyl-7-(propan-2-ylidene)azulen-5(1H)-one,original curcumol and curcumin in Curcumae Aromaticae Radix,using curdione as intermal reference substance,and the relative correction factor (RCF) of the four components was determined by HPLC with good reproducibility.Their contents in 10 batches of samples,collected from different areas,were determined by both external standard method and QAMS.Result No significant differences were found in the quantitative results of four compounds in 10 batches of Curcumae Aromaticae Radix determined by external standard method and QAMS.Conclusion It is feasible and suitable to evaluate the quality of Curcumae Aromaticae Radix by QAMS.

OBJECTIVETo investigate the association of single nucleotide polymorphisms (SNPs) in VEGF gene with the risk of endometriosis and adenomyosis.

METHODSGenotypes were analyzed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method in 344 endometriosis patients, 174 adenomyosis patients, 360 frequency-matched control women of endometriosis and 199 frequency-matched control women of adenomyosis.

RESULTSNo significant difference was found in allele frequencies and genotype distributions of the -460C/T polymorphism between patients (endometriosis and adenomyosis) and control women (all P value > 0.05). However, there were significant differences in genotype and allele distributions of the VEGF -1154G/A polymorphism between patients (endometriosis and adenomyosis) and control women (all P value < 0.05). The genotype frequencies of the VEGF -1154 AA, GA, and GG in endometriosis patients and control women were 1.7%, 28.8%, 69.5% and 5.8%, 32.8%, 61.4%, respectively; and the A and G allele frequencies in the two groups were 16.1%, 83.9% and 22.2%, 77.8%, respectively. The genotype frequencies of the VEGF -1154 AA, GA, and GG in adenomyosis patients and control women were 2.9%, 23.6%, 73.6% and 7.0%, 34.2%, 58.8%, respectively; and the A and G allele frequencies in the two groups were 14.7%, 85.3% and 24.1%, 75.9% respectively. Compared with GA+ AA genotype, GG genotypes could significantly increase the risk of endometriosis (OR:1.43,95%CI:1.05-1.96) and adenomyosis (OR:1.95,95%CI:1.26-3.03).

CONCLUSIONThe VEGF -1154G/A polymorphism was associated with susceptibility to endometriosis and adenomyosis, and the GG genotype could significantly increase the risk of developing endometriosis and adenomyosis. However, the VEGF -460C/T polymorphism was not associated with susceptibility to endometriosis and adenomyosis in the population studied.

5' Untranslated Regions ; Adult ; Biophysical Phenomena ; Endometriosis ; genetics ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; genetics ; Genotype ; Humans ; Polymorphism, Genetic ; Polymorphism, Single Nucleotide ; Promoter Regions, Genetic ; Vascular Endothelial Growth Factor A ; genetics

OBJECTIVETo study the cytotoxic effect of allogenetic natural killer (NK) cells in vitro on human CD34+ acute myelogenous leukemia cells.

METHODSCD34 expression on acute myelogenous leukemia KG1a cells was detected by flow cytometry. KG1a cells were co-cultured at different effector-to-target (E:T) ratios with NK cells isolated from 5 healthy individuals using magnetic cell sorting. Lactate dehydrogenase (LDH) release assay was employed to examine the cytolysis of KG1a cells in the co-culture, and the inhibition rate of the KG1a cell colony formation in methylcellulose was determined with K562 cells sensitive to NK cells as the control.

RESULTSA expression rate as much as (98.0-/+1.1)% was detected for CD34 antigen on KG1a cells, and the isolated NK cells (CD3(-)CD16+CD56+ cells) had a purity of (93.2-/+3.7)% after magnetic cell sorting. Allogenetic NK cells exhibited obvious cytotoxicity and colony inhibition in vitro against KG1a cells at different E:T ratios, and the effects were significantly enhanced as the E:T ratios increased (P<0.05). At the same E:T ratio, the cytotoxicity and colony inhibition rate of allogenetic NK cells against KG1a cells was lower than those against K562 cells (P<0.05).

CONCLUSIONAllogenetic NK cells exhibit obvious cytotoxicity and colony formation against CD34+ acute myelogenous leukemia cells.

Antigens, CD34 ; immunology ; Coculture Techniques ; Cytotoxicity, Immunologic ; Flow Cytometry ; Humans ; K562 Cells ; Killer Cells, Natural ; immunology ; Leukemia, Myeloid, Acute ; immunology

OBJECTIVETo construct a recombinant adenovirus of survivin vector and provid valuable reference for gene therapy of laryngeal cancer.

METHODSThe survivin gene was cloned by PCR. After confirmation by enzyme restriction analysis and sequencing, the gene and the adenovirus vector were recombined together to construct the recombinant adenovirus vector. The recombinant adenovirus vector was confirmed via both sequencing and digestion restriction analysis, and then linearized and transfected into the HEK 293 cell line to generate recombinant adenovirus.

RESULTSThe sequence analysis demonstrated that the survivin gene sequence was the same as published in the literature, suggesting that a recombinant adenovirus vector has been successfully constructed.

CONCLUSIONSA survivin recombinant adenovirus has been successfully constructed.

Adenoviridae ; genetics ; Genetic Vectors ; HEK293 Cells ; Humans ; Inhibitor of Apoptosis Proteins ; genetics ; metabolism ; Plasmids ; Polymerase Chain Reaction ; Recombinant Fusion Proteins ; genetics ; metabolism ; Transfection

Two Rotavirus G9P[8] strains (LL52696 and LL52727) were recognized during a sentinel-based survey in Lulong, China. Phylogenetic analysis of the VP7 gene showed that both strains isolated constituted a divergent genetic cluster distinct from the other G9 strains isolated in China. Analysis of VP4, VP6, and NSP4 genes revealed that these strains were closely related to Lulong strains. We hold that two strains were reassortant between G9 and Lulong predominant strains.
Amino Acid Sequence ; Antigens, Viral ; chemistry ; genetics ; Base Sequence ; Capsid Proteins ; chemistry ; genetics ; Glycoproteins ; chemistry ; genetics ; Humans ; Phylogeny ; Rotavirus ; classification ; genetics ; Toxins, Biological ; chemistry ; genetics ; Viral Nonstructural Proteins ; chemistry ; genetics