Four kinds of ionic liquids were adopted to analyze the content of rubimaillin and alizarin in Rubia cordifolia roots with ultrasonic-assisted extraction coupled with HPLC. The chromatographic column, Purospher star RP-C18 (4.6 mm x 250 mm, 5 microm), was used. Methanol and 0.4% acetic acid-water as mobile phase with flow rate at 0.85 mL min(-1), gradient elution, detection wavelength at 250 nm, chromatographic column temperature was controlled at room temperature. The result showed that rubimaillin and alizarin had the highest extraction yield when the [ HMIM] PF6methanol solution concentration of 0.6 mol x L(-1) as extraction solvent and the conditions were solid-liquid ratio of 1:80 (g x mL(-1)). Under the optimal extraction conditions, the content of alizarin from 0.01 to 0.04 microg showed a good linearity (r = 0.9999), the average recovery was 97.12%, the content of rubimaillin from 0.41 to 1.35 microg showed a good linearity (r = 0.9999), the average recovery was 98.10%. This experiment adopted environmentally friendly reagent as extraction solvent, the extraction efficiency was improved, and the environmental pollution caused by organic solvent was avoided, the harm of human body aslo was reduced. This method was simple and reliable, its repeatability was also very good, which had an important significance in the study of traditional Chinese medicine active ingredient extraction methods.
Anthraquinones ; analysis ; Chromatography, Reverse-Phase ; methods ; Ionic Liquids ; chemistry ; Pyrans ; analysis ; Rubia ; chemistry ; Ultrasonics

BACKGROUND:Gemcitabine hydrochloride is a water-soluble anticancer drug that induces apoptosis in tumor cells, but it has an excessive release in vivo. OBJECTIVE:To evaluate the effect of poly(lactic acid) (PLA) electrospun fiber membranes carrying gemcitabine hydrochloride on the growth of human osteosarcoma cell lines MG-63. METHODS:PLA electrospun fiber members with (experimental) or without (control) gemcitabine hydrochloride were fabricated and characterized. Two kinds of fiber membranes were immersed in low-glucose DMEM medium, and the supernatants were collected in the two groups at 3, 5, 7 days, respectively. Passage 5 human osteosarcoma cell lines MG-63 were inoculated into 96-well plates containing low-glucose DEME with 15％ fetal bovine serum, and divided into seven groups. Groups 1-3 were cultured in the experimental supernatants of 3, 5, 7 culture days, and groups 4-6 were cultured in the control supernatants of 3, 5, 7 culture days, respectively. The remaining group acted as the negative control with no supernatant. Thereafter, cell counting kit-8 was used to detect cell proliferation, and RT-PCR was used to measure expression of Bcl-2 and Bax at 3 days of culture. RESULTS AND CONCLUSION:(1) No obvious particle was found on the smooth and even surface of the fiber members in the experimental and control groups. There was no significant difference in fiber diameter, contact angle and tensile strength between the two kinds of fiber membranes. (2) The results of cell counting kit-8 showed that compared with the negative control group, the supernatant released from the control group had no effect on the MG-63 proliferation at different time points, while the supernatant released from the experimental group could inhibit the MG-63 proliferation at different time points (P<0.05), and the inhibitory effect became more and more obvious with the prolongation of release time. (3) RT-PCR findings showed that compared with the control group, the supernatant released from the experimental group could increase Bax mRNA expression and decrease Bcl-2 mRNA expression at the same time point. To conclude, the PLA electrospun fiber membranes carrying gemcitabine hydrochloride can sustainably inhibit MG-63 proliferation and promote cell apoptosis.

In order to confirm the role that the 49th amino acid residue plays in enzymatic inactivity of Glutamine 49 phospholipase A2(Gln49-PLA2),site-directed mutagenesis of its 49th amino acid gene codon was conducted using PCR.Aspartic acid 49 phospholipase A2(Asp49-PLA2-Q49D-PLA2),the mutant of Gln49-PLA2 was expressed in E.coli with pET32a+ vector.The fusion protein,expressed as inclusion body,after being denatured,was on-column refolded and purified by immobilized metal affinity chromatography(IMAC),and then cleaved by Factor Xa.The mature Q49D-PLA2 mutant was obtained by Hitrap SP cation exchange and Superdex 75 gel filtration chromatography,with the recovery rate of 1.3%,and the specific activity of the mature Q49D-PLA2 mutant was 72 U/mg.It has been demonstrated that the 49th glutamine amino acid residue is the main reason in enzymatic inactivity of Gln49-PLA2 and the results are helpful for denatured protein refolding,especially in rich disulfide bonds conditions.

Adult ; Biomarkers ; blood ; Case-Control Studies ; Female ; Granzymes ; blood ; Hepatitis B ; blood ; immunology ; Humans ; Male ; Middle Aged ; Perforin ; blood ; T-Lymphocytes, Cytotoxic ; metabolism

Objective To look into the current distribution of iodine deficiency area in Shandong province and to guide the re-defined iodine deficiency area and to supplement iodine scientifically. Methods In 2008, 100 iodine deficiency counties(cities, districts), designated in Shandong province's "to supplement iodized salt to eliminate the hazard of iodine deficiency management regulations", were selected in the study. One to three samples were collected from water source which was used by the majority of local residents in the 100 iodine deficiency places and iodine concentration was tested by As3+-Ce4+ catalyzing spectrophotometry. Results A total of 65 716 water samples were collected. Sample recovery efficiency reached 99.8%(65 572/65 716). The median water iodine was 5.57 μg/L, with 82.05%( 1097/1337 ) of the township(town) met criteria for the classification of iodine deficiency areas(water iodine ＜ 10 μg/L), 17.43%(233/1337) of the township (town) water iodine moderate(water iodine 10 - 150 μg/L), and 0.52%(7/1337)of the township(town) should be defined high iodine areas(water iodine ＞ 150 - 300 μg/L). Conclusions The iodine deficiency areas should be redefined because water iodine concentrations of iodine deficiency areas have changed. We suggest that the smallest place to supply salt with different range of iodine content is set to the township(town).

Objective To explore the effect of family support service intervention on improving the rehabilitation of patients with severe mental disorders in community and the mental health status and family burden of family members. Methods Using multi-stage random sampling method, 100 patients who met the diagnostic criteria of severe mental disorders were randomly selected from two communities, and then 100 patients who met the diagnostic criteria of severe mental disorders were randomly matched according to gender, age and diagnosis in other communities into the control group. The control group and intervention group were set up strictly according to the inclusion criteria of patients and their families. Results The average age of the 200 groups was (48.27±12.67) years, and the average age of the family members was (63.61±13.19) years. After intervention, the activity dailyliving scale (ADL) scores of the control group were higher than those of the intervention group at all time points (all P<0.05). After intervention, the social disability screening schedule (SDSS) scores of the control group were higher than those of the intervention group at all time points (all P<0.05). After intervention, there was no significant difference in the morningside rehabilitation status scale (MRSS) score between the intervention group and the control group at all time points (all P>0.05). After intervention, the SCL-90(self-reporting inventory) scores of the mental health of the family members in the control group were higher than those in the intervention group at all times (all P<0.05). After intervention, the family burden scale of diseases (FBS) scores of the control group were higher than those of the intervention group at all time points (all P<0.05). Conclusions The intervention measures did improve the rehabilitation effect of severe mental disorder patients in community and the psychological and family burden of family members. A professional family support service team should be established.

OBJECTIVETo observe the role of interleukin (IL) 21 alone, IL12 alone, and IL21 plus IL12 for inducing the antitumor activity of peripheral blood mononuclear cells (PBMCs) in patients with endometrial cancer.

METHODSPBMCs were isolated from peripheral blood in patients with endometrial cancer in vitro, and kept the culture with low-level IL2. IL2-stimulated PBMCs were cocultured under different conditions (with anti-IL21 antibody, IL21 alone, IL12 alone, or IL21 plus IL12) for 72 h. The cytotoxicity of PBMCs was then examined by lactate dehydrogenase(LDH) released assay. CD4(+) CD25(+) FOXP3(+) regulatory (Treg) cell and CD4(+) IL17A(+) T-helper (Th17) cell proportion were determined with flow cytometry. Cell proliferation and apoptosis were measured by cell counting kit-8(CCK-8)assay and flow cytometry, respectively.

RESULTSIn comparison to control group, both IL21 and IL12 significantly enhanced the cytotoxicity of PBMCs. The IL21 plus IL12 group had superior effect to IL21 alone and IL12 alone. IL21 and IL12 significantly decreased the percentages of Treg cells and the rate of PBMCs apoptosis. IL21 or IL12 had no significant effect on the differentiation of Th17 cells and the proliferation of PBMCs.

CONCLUSIONSIL21 and IL12 can enhance the cytotoxicity of PBMCs in patients with endometrial cancer, which can be further strengthened with treatment of IL21 plus IL12. Such effects may be achieved by inhibiting the differentiation of Treg cells and the apoptosis of PBMCs, but not by the differentiation of Th17 cell.

Adult ; Aged ; Apoptosis ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Endometrial Neoplasms ; immunology ; pathology ; Female ; Humans ; Interleukin-12 ; pharmacology ; Interleukins ; pharmacology ; Leukocytes, Mononuclear ; drug effects ; immunology ; pathology ; Middle Aged ; T-Lymphocytes, Regulatory ; immunology

Objective To investigate the effect of overexpression of α 2,3-sialyltransferase (ST3Gal Ⅲ) on abilities of invasion in breast cancer cells MDA-MB-231.Methods ST3Gal Ⅲ gene lentiviral expression vector was transfected into MDA-MB-231 cells.Transfected cells were divided into 3 groups:blank group (Mock cells,M),control group (Parent cells P),experimental group (ST3Gal Ⅲ ST3).The expression of ST3Gal Ⅲ mRNA and protein were detected by Quantitative Real-time PCR(RT-PCR) and Western blot.The content of α2,3-sialic acids was analyzed by flow cytometry.The ability of migration and invasion in MDA-MB-231 cells were detected by cell adhesion assay and Transwell assay,respectively.Results The expression levels of ST3Gal Ⅲ mRNA and protein examined by RT-PCR and Western blot in control group and experimental group were 0.91 ± 0.04 and 1.85 ± 0.08 respectively.The expression levels in experimental group were 1.84-fold and 1.76-fold higher than control group(P ＜ 0.05).The amount of SLeX on the cell surface in experimental group was 92.86 ± 4.41 and was 1.61-fold higher than control group,which were significantly increased compared with that in the blank group and control group (P ＜ 0.05).OD570nm values of the adhesion assay in the in the blank group,the control group and the experimental group were 0.32 ± 0.02,0.29 ± 0.01 and 0.46 ±0.03,respectively,while those of the invasion assay were 0.93 ±0.02,0.81 ±0.01 and 1.46 ±0.13,and there was significant difference between the experimental group and the control group (P ＜ 0.05).Conclusion Overexpression of ST3Gal Ⅲ gene could enhance the invasion ability of MDA-MB-231 cells.

Objective To investigate the frequency of the breast cancer related lymphedema and its effects on the health outcomes of breast cancer survivors.Methods It was a cross-sectional study and 301 female breast cancer survivors were enrolled in the study.Circumference measurement were chosen to diagnose the lymphedema.Quality of life Questionnare-Core30 designed by European Organization for Research and Treatment of Cancer and Disabilities of Arm,Shoulder and Hand Scale were administered to assess the quality of life and upper limb function which were calculated as the main outcome variables.Results The incidence of the breast cancer related lymphedema was 15.0％.The patients with breast cancer related lymphedema had lower mean values in physical functioning and role functioning and the differences were statistically significant (P ＜0.05),respectively.Patients with breast cancer related lymphedema had higher DASH scores and the difference was statistically significant (P ＜ 0.05).Conclusions Breast cancer related lymphedema can not only decrease breast cancer survivors quality of life but also impair their upper limb function.It implied that breast cancer related lymphedema is a question which needed to be paid more attention.

AIMTo study the effect of curcumin on the expression of prostate specific antigen (PSA).

METHODSAXSYM system-chemical luciferase method was used to examine the content of PSA in prostate cancer cell lines, LNCap after treated with different doses of curcumin. pGL3-PSA luciferase expression vector, containing 640 bp DNA of PSA gene 5' promoter region was constructed and transfected into LNCap cell with lipofectin. Through detecting the activity of luciferase, the effect of curcumin on the promoter of PSA was studied. Western blotting was used to detect expression of androgen receptor (AR) in LNCap cell with different concentrations of curcumin.

RESULTSThe expression of PSA was inhibited and activity of luciferase was reduced by curcumin. There was also significant difference in AR expression as shown by Western blotting experiment after treatment of different doses of curcumin.

CONCLUSIONThrough inhibiting AR expression, curcumin reduced the function of PSA promoter and inhibited PSA protein expression.

Antineoplastic Agents ; pharmacology ; Cell Line, Tumor ; Curcumin ; pharmacology ; Humans ; Luciferases ; metabolism ; Male ; Promoter Regions, Genetic ; drug effects ; Prostate-Specific Antigen ; genetics ; metabolism ; Prostatic Neoplasms ; immunology ; metabolism ; pathology ; Receptors, Androgen ; metabolism