Aim To study preparation of lutein from the extraction Marigold flowers by biotransformation.Methods Screen bacterium for lutein esters biotransforming by studying on the biotransformation of lutein esters by Penicillium on tabulate revulsive culture medium.Porule of and Aspergillus niger Penicillium could transform lutein esters into lutein,but biotransformation of Penicillium was better than Aspergillus niger.Porule of Pleurotus ostreatus could not grow on revulsive culture medium,but Pleurotus ostreatus could grow on the culture medium and transformed lutein esters into lutein.Research of shaking flash fermentation of Penicillium for trans-forming lutein esters.When pH is 6.0,quantity of incubation is 2 bacteria plugs and fermentation time is 12～24 hours,the transformation is completed and the content of lutein is 91.6%(HPLC)。Result Prolonging the transformation time will improve the percent of lutein but the content of lutein decreased.Conclusion Biological transrormation can be used from calendula officinacis extract preparation.

Objective To develop a rapid and sensitive method for the determination of bisphenol A in drinking water by liquid chromatography-electrospray tandem mass spectrometry. Methods Bisphenol A was extracted from the drinking water sample by a SEP-PAK C18 column,eluted with methanol and concentrated with a rotary evaporator. The extract was analyzed by liquid chromatography-tandem mass spectrometry with electrospray ionization. Results The calibration curve of bisphenol A was linear in the range of 5 -100 ng/ml,the linear equation was y =6 796.61x -8 655.64 and the correlation coefficients of linear calibration curve was 0.999 2. The limit of detection (S/N=3) and the limit of quantification (S/N=10) of bisphenol A were 0.007 5 ng/ml and 0.025 ng/ml,respectively. The rates of recovery of bisphenol A were from 83.46% to 94.00% and the relative standard deviation was 3.06% -4.60%. Conclusion The method is simple,rapid,accurate and is applicable to the qualitative and quantitative determination of bisphenol A in drinking water.

Objective To investigate the effects of erythropoietin(EPO)on the function of glomerular endothelial cells in rats with chronic renal failure(CRF). Methods The CRF model was established by a two stage 5/6 nephrectomy procedure in rats.Experimental rats were randomly divided into four groups:sham operation group (control group),CRF group,CRF rats treated with 30 U/kg EPO(low-dosage group)and with 50 U/kg EPO (high-dosage group).CRF rats received EPO by hypodermic injection for 6 weeks and then were sacrificed.Serum creatinine(Scr),blood urea nitrogen fBUN),urine protein,haematoglobin (Hb) and blood pressure were measured.The renal morphologie changes were evaluated on periodic acid-schiff (PAS) stained sections.The CD34 and CD31 expressions in glomerulus were detected by immunohistochemistry method.The mRNA of endothelin 1(ET-1),endothelial nitric oxide synthase (eNOS) and vascular endothelial growth factor (VEGF) were detected by RT-PCR. Results The expressions of CD34 and CD31 protein in glomerulus,and the expressions of eNOS and VEGF roRNA in renal tissue were higher in EPO treatment group than those in CRF model group(all P<0,05).The expression of ET-1 mRNA in renal tissue was lower in EPO treatment group than that in CRF model group.In addition,the Scr,BUN,urine protein and blood pressure in EPO treatment group were significantly lower than those in CRF model group (all P<0.05).Haematoglobin in EPO treatment group was higher than that in CRF model group (P<0.05).Reanl pathological injury wss improved by EPO treatment in dose-dependent manner. Conclusion EPO can ameliorate renal pathological injury and renal function in rats with chronic renal failure,maybe through promoting the renovation of glomerular capillary endothelium and improving the function of glomerular endothelial cells.

Objective To investigate the effects of erythropoietin (EPO) on the number and function of peripheral blood endothelial progenitor cells (EPCs) from rats with chronic renal failure (CRF). Methods The model of chronic renal failure was established by a two-stage 5/6nephrectomy procedure in rats. Experimental rats were randomly divided into four groups (n =7,respectively): sham operation group, CRF group, CRF rats treated with 30 U/kg EPO (low-dosage group) and with 50 U/kg EPO (high-dosage group). CRF rats were given EPO by hypodermic injection for 6 weeks, then EPCs were isolated by density gradient centrifugation from peripheral blood mononuclear cells. The ability of cell proliferation, adhesion and vasculogenesis in vitro was further observed. Results Compared to sham operation group, the ability of cell proliferation,adhesion and vasculogenesis in vitro in CRF rats was remarkably decreased (P＜0.05, respectively).Such ability was promoted significantly in dose-dependent manner by EPO treatment (P＜0.05,respectively). Conclusion EPO can improve the number and ability of endothelial progenitor cells from rats with chronic renal failure.

Objective To investigate the differentiation of rat bone marrow mesenchymal stem cells (MSCs) to renal tubular epithelial-like cells under different conditions. Methods MSCs were obtained from rat marrow. MSCs were isolated by gradient density centrifugation and plastic adherence and then purified. Surface markers were identified with flow cytometry after amplification in vitro. The purified MSCs of the third passage were cultured respectively as follows: (1) control group: DMEM medium with fetal bovine serum(FBS). (2) all-trans retinoic acid (ATRA) group: DMEM medium with FBS, ATRA and ischemic reperfusion-injured kidney tissue homogenate. (3)combination group: DMEM medium with FBS, ATRA, ischemic reperfusion-injured kidney tissue homogenate, epidermal growth factor (EGF) and bone morphogenetic protein 7 (BMP-7). After 7 days, the MSCs were collected for alkaline phosphatase (AKP) staining, cytokeratin-18 and E-cadherin immunocytochemical analysis. Results The positive rates of the third passage MSCs in CD44, CD90 and CD29 were 97.8%±0.9%, 96.8%±1.4% and 97.6%±2.4%,respectively, but in CD11b/c and CD34 were only 13.2%±0.6% and 1.2%±0.5%. The MSCs in control group were spindle. The MSCs in ATRA group were round and elliptic. The MSCs in combination group became cobblestone-like cells after 7 days. AKP staining showed that tubular epithelial-like cells from MSCs in control group were negative, some above cells in ATRA group were positive and number of above cells increased in combination group. Compared with negative control group, the ratios of cytokeratin-18 positive cells in ATRA group and combination group were respectively increassed by 29.47%±1.08% and 47.52%±2.13% (all P<0.05), the ratios of E-cadherin positive cells in ATRA group and combination group were respectively increased by 14.88%±2.46% and 36.15%±1.13% (all P<0.05). Conclusion MSCs may differentiate by renal tubular epithelial-like cells under the induction of ischemic reperfusion-injured kidney tissue homogenate and ATRA in vitro, which are further differentiated under the combined induction of EGF and BMP-7.

Objective To observe the effects of bone marrow?derived mesenchymal stem cells (BMSC) on glomerular podocyte injured by lipopolysaccharide (LPS) and the expression of related protein. Methods Podocytes are divided into control group, BMSC group, LPS group and LPS plus BMSC group. After 24 hours of intervention, observing each experimental group podocyte form under inverted phase contrast microscope;detecting the expressions of mRNA and protein of nephrin, CD2AP, synaptopodin, and TRPC6 by RT?PCR and Western?blot. Results Compared with control group, expressions of nephrin, CD2AP, and synaptopodin in LPS group decreased (P<0.05) while that of TRPC6 increased (P<0.05); compared with LPS group, expressions of nephrin, CD2AP, and synaptopodin in LPS+MSC group increased (P<0.05) while that of TRPC6 decreased (P<0.05). Conclusion BMSC may relieve LPS?induced podocyte injury.

Objective To evaluate the clinical efficacy and safety of capecitabine continuous maintenance in treatment of re-currence of triple-negative breast cancer,which had responded to combined chemoradiotherapy.Methods The triple-negative breast cancer was defined as negative of ER,PR and HER2.A total of 46 patients of triple-negative breast cancer were divided into the treatment group (23 patients)and the control group (23 patients).The treatment group was given capecitabine continuous mainte-nance after 6 cycles of chemotherapy.The control group was given 6 cycles of combined chemotherapy only.The clinical efficacy and safety was evaluated between the two groups.Results The PR,PD,RR,and DCR in the treatment group were significantly higher than those in the control group(P <0.05).The acute toxic effects (except for hand-foot syndrome)were similar in two group(P >0.05).Conclusion Capecitabine continuous maintenance after combined chemoradiotherapy in treatment of recurrence of triple-neg-ative breast cancer is more effective,lower toxicity and tolerable.

OBJECTIVETo explore the association of KIR-HLA gene polymorphism with chronic myeloid leukemia (CML) among ethnic Hans from southern China.

METHODSA total of 172 adult CML patients and 480 unrelated healthy controls were screened for the presence of KIR with sequence-specific primers-PCR (PCR-SSP) and sequence-based typing (SBT) of HLA-A, -B and -C loci. Polymorphisms of the KIR-HLA system were analyzed at 4 levels, and the frequencies of KIR framework genes and KIR profiles, classⅠHLA ligands, matched KIR+HLA pairs and KIR-HLA compound profile were compared between the two groups. P values were calculated using SPSS 13.0 software.

RESULTSFor the CML group, the frequencies of HLA-C2 ligand, 2DL1+HLA-C2 pair and HLA-B Bw4-80I were significantly lower than those of the control group, suggesting a protective effect against CML (HLA-C2: OR=0.386, 95%CI:0.240-0.620, P<0.01; 2DL1+HLA-C2: OR=0.316, 95%CI:0.191-0.525, P<0.01; HLA-B Bw4-80I: OR=0.576, 95%CI:0.384-0.862, P<0.01). The frequencies of KIR2DL1 ligand (HLA-C2) and KIR3DL1 ligand (HLA-B Bw4-80I) in the CML group were significantly lower than that of the control group, suggesting that the HLA-C2 and HLA-B Bw4-80I expression is probably decreased in the CML patient group, which led to reduced inhibitory signal and enhanced activating signal of KIR2DL1and/or KIR3DL1NK cells. Notably, the frequency of KIR-HLA compound profiles ID2 (KIR AA1-HLA-C1/C1-Bw6/Bw6-A3/11) in CML patients significantly increased in the CML patient group compared with the control group, suggesting that the KIR-HLA compound profiles ID2 may be a risk factor for CML (OR=2.163, 95%CI 1.198-3.906, P<0.01).

CONCLUSIONAbove analysis has identified certain protective and risk factors for CML from the KIR-HLA system, which may provide a clue for the pathogenesis of leukemia and development of individualized immune therapy.

Asian Continental Ancestry Group ; genetics ; China ; Gene Frequency ; Genetic Predisposition to Disease ; ethnology ; genetics ; Genotyping Techniques ; HLA Antigens ; genetics ; HLA-A Antigens ; genetics ; HLA-B Antigens ; genetics ; HLA-C Antigens ; genetics ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; ethnology ; genetics ; Odds Ratio ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Protein Isoforms ; genetics ; Receptors, KIR ; genetics ; Risk Factors

Objective: To investigate the effect and mechanism of renal fibrosis after macrophage depletion in C3-deficient unilateral ureteral obstruction mice. Methods: Renal interstitial fibrosis model was established by unilateral ureteral obstruction (UUO) in male C3-deficient mice and age-matched C57BL/6 WT mice (8-12 weeks of age). Mice were randomly divided into 4 groups, including sham operation in wild type group (WT/sham) (n=18), UUO operation in wild type group (WT/UUO) (n=18), sham operation in C3-deficient group (C3KO/sham) (n=18), and UUO operation in C3-deficient group (C3KO/UUO) (n=18). The expression of complement C3 was detected by immunohistochemical staining and renal interstitial macrophages were assessed by immunofluorescence staining. Tubulointerstitial fibrosis was observed by both HE staining and Masson staining after 14 days of UUO. Collagen accumulation and score of tubulointerstitial injury were obtained. Wild type and C3-deficient UUO mice were treated by liposome clodronate in early or late stage respectively and then interstitially infiltrated macrophages and renal fibrosis were analysed. Mice were sacrificed randomly at 3,7,14 days after UUO and obstructed kidneys were collected. Macrophage phenotype was detected by double-labeling immunofluorescence with F4/80 and iNOS for the M1, F4/80 and CD206 for the M2 macrophage subpopulation. iNOS, Arg-1 and CD206 were also detected by western blot. Results: C3 deficient mice exhibited attenuated renal fibrosis, reduced collagen accumulation and tubulointerstitial injury score compared with WT mice (P<0.01). Meanwhile, macrophage depletion in early or late stage of UUO reduced renal fibrosis in WT mice, but had no effect on C3-deficient UUO mice. Decreased accumulation of M1 macrophages and expression of iNOS, increased accumulation of M2 macrophages and expression of Arg-1, CD206 were found in C3 deficient mice compared with WT mice in early stage of UUO (P<0.01). Conclusion Renal fibrosis is not reduced after depletion of macrophages in C3 deficient UUO mice due to the altered macrophage polarization.

OBJECTIVE: To study application of sternocleidomastoid myoperiosteal flap to reconstruction of the defect of cervical trachea. METHOD: Thirteen hospitalized patients with malignant neoplasm invading cervical trachea and primary tracheal carcinoma were analyzed retrospectively, the sternocleidomastoid myoperiosteal flap was applied to repair the defect of tracheal wall after resection of the neoplasm. RESULT: Among 10 tracheotomy patients, 9 cases were decannulated from 1 month to 5 months. One case needs cannulation because of tracheal stenosis. Three patients with no tracheotomy had no dyspnea after operation. CONCLUSION The sternocleidomastoid myoperiosteal flap is an ideal transplant for cervical tracheal reconstruction.
Aged ; Female ; Humans ; Male ; Middle Aged ; Pectoralis Muscles ; transplantation ; Periosteum ; transplantation ; Reconstructive Surgical Procedures ; methods ; Retrospective Studies ; Surgical Flaps ; Trachea ; pathology ; surgery