BACKGROUND:Recently,some overseas in vitro researches and animal models confirmed that donor leukomonocyte cultured by fludarabine can significantly reduce acute graft-versus-host disease,but the mechanism of action is not clear.OBJECTIVE:To explore immunophenotype changes of donor leukomonocyte cultured with fludarabine in vitro.DESIGN,TIME AND SETTING:The observational cytology experiment was performed at the Department of Hematology,First People's Hospital from July 2007 to January 2008.MATERIALS:Leukomonocytes were collected from four hour healthy persons as donors of allogenic peripheral blood hematopoietic stem cell transplantation.Fludarabine was offered by Schering AG,Germany.METHODS:Peripheral blood hematopoietic stem cells collected by COBO SPECTRA were washed with RPMI1640 twice.These cells were cultured in RPMI1640 containing 0.10 volume fraction of fetal bovine serum till cell density of 2?109 L-1.1 mL of hematopoietic stem cells were washed with phosphate-buffered saline,and added CD3,CD4,CD8,CD44mAb 10 ?L,and added Mouse Ig-G1-FITC/PE as control group.These cells were attenuated with phosphate-buffered saline after incubated for 30 minutes at a low temperature,then detected by flow cytometry.Other cells were put in 24-well cultivate plate,added 8 mmol /L fludarabine as a final concentration for 24 hours(5% CO2,37 ℃).MAIN OUTCOME MEASURES:Studying immunophenotypic changes of donor leukomonocytes by flow cytometry.RESULTS:There was no significant change in the ratio of CD4+CD44+T cells before or after donor leukomonocytes cultured with fludarabine for 24 hours,and proportion of CD8+CD44+T cells declined significantly(P

Objective To study the chemical constituents of Acanthus ilicifolius. Methods Compounds were isolated and purified on silica gel,SephadexLH -20 and HPLC,and their chemical structures were elucidated by spectral evidence,physical and chemical analyses. Results and Conclusion Nine compounds,named as stigmasta-4-en-3-one(1),stigmasta-4,22-diene-3-one(2),campest-4-en-3-one(3),tigmasta-4-en-3,6-dione(4),Stigmasta-4,22-diene-3,6-dione(5),Stigmasta-4,22-diene-6?-ol-3-one(6),6?-hydroxyl-stigmasta-4-en-3-one(7),3?-hydroxyl-stigmasta-5,22-diene-7-one(8),3?-hydroxyl-stigmasta-5-en-7-one(9),respectively,were obtained from the petroleum ether soluble portion in the methanol extracts of Acanthus ilicifolius. Among them,compounds 4~8 were reported from this plant for the first time,compounds 2,4 and 9 showed moderate activity against selected tumor cell lines.

AIM:To establish the HPLC-fingerprint of the water-soluble constituents of Salvia miltiorrhizae Bge.on 18 batches from 9 areas in China. METHODS: Samples of Salvia miltiorrhizae Bge.from different areas were determined by Agilent 1100 DAD-HPLC on Sino Chrom ODS-BP column(250 mm?4.6 mm,5 ?m),gradient elution with methanol-water(0.7%H_3PO_4) as mobile phase,column temperature was at 30℃,flow rate was 1 mL/min,wavelength was at 280 nm,and the inject volume was 20 ?L. RESULTS: The HPLC-fingerprints of the water-soluble constituents from Salvia miltiorrhizae Bge.,including 12 mutual peaks,were developed after determination of 10 batches of drugs according to SPSS analysis.,and in which,3 characteristic components were recognized. CONCLUSION: The separation effect of peaks in fingerprints is better and the characteristic is also obvious.The RSD values of precision and reproducibility are less than 5%.This method can be used for the quality control of Salvia miltiorrhizae Bge.

Objective To explore the impact of recombinant human hepatocyte growth factor (rhHGF) in Celsior (CS) solution on the expression of INF-γ, IL-4 and IL-10 in a rat liver transplan-tation model. Methods After flushed with CS solution with addition of rhHGF (experimental group) or saline (control group), NHBD livers were stored at 4℃; for 16 h.then they were transplanted using the two-cuff technique with arterial reconstruction. The serum levels of INF-γ, IL-4 and IL-10 at lh after reperfusion were detected using ELISA. The INF-γ, IL-4 and IL-10 mRNA in the corresponding liver tissue were determined by RT-PCR. The 7-day survival rate was calculated and the histopatho-logical examination results were analyzed by hematoxylin and eosin staining. Results Compared with the control group, the experimental group showed lower INF-γ level and higher IL-4 and IL-10 levels in serum at 1 h after reperfusion (P<0. 05). The level of INF-γ mRNA in liver tissue was significant decreased at 1 h after reperfusion (P<0. 05) , and the level of IL-4 and IL-10 mRNA was significantly increased in the experimental group (P<0. 05). In experimental group, recipients got a better survival rate and histopathological examination showed a well-preserved hepatic architecture without hepatocyte necrosis, milder sinusoidal and portal congestion. Conclusion Adding exogenous rhHGF in CS solu-tion can protect NHBD livers from ischemia-reperfusion injury and prolong the survival in rats, which might be due to down-regulation of TNF-γ and up-regulation of IL-4 and IL-10.

Objective To investigate the chemical constituents of gorgonian Muriceides collaris. Methods The compounds were isolated and purified by silica gel column,Sephadex LH-20 column chromatography and HPLC. Their chemical structures were identified by physicochemical analysis,spectroscopic analysis,and comparison with the data of literatures. Results From the CH3OH soluble extract of Muriceides collaris,nine compounds were isolated,and their structures were determined as cholesterol (1),batyl alcohol (2),benzoic acid (3),uracil (4),thymine (5),2'-deoxyuridine (6),2'-deoxythymidine (7),thymidine (8) and 2'-deoxyadenosine (9),respectively. Conclusion All compounds were obtained from the species Muriceides collaris for the first time.

Objective To assess the effectiveness of repeated trans-cranial magnetic stimulation ( rTMS) in relieving post-stroke depression ( PSD). Methods PubMed, Embase, the Cochrane library, Web of Science, CNKI, WANFANG, and VIP were searched for reports of randomized, controlled trials of rTMS treatment of PSD published before June 2015. Crude standardized mean differences ( SMDs) and odds ratios with 95% confidence in-tervals ( CIs) were calculated for depression intensity and effectiveness rate after treatment using random or fixed effects models. Results Twenty-four studies involving 856 rTMS-treated patients and 802 control patients were in-cluded in the meta-analysis. The results showed that compared with the control group, PSD patients showed significant reductions in depression after rTMS treatment ( SMD=-1.36;95% CI-1.6 to-1.12;P≤0.05) . The total effective-ness rate in the treated group was 85% with a reduction in NIHSS score ( SMD=-0.82;95% CI-1.2 to-0.44;P≤0.05) . Subgroup analysis showed that neither the frequency of rTMS stimulation, the site stimulated, nor time after stroke had a significant influence on the effectiveness of rTMS. Additionally, a few studies reported adverse reactions after rTMS. Conclusion rTMS appears to be a safe and effective therapy for PSD. Further well-controlled trials may elucidate the mechanism underlying the placebo effects of the sham rTMS observed among PSD patients.

This study was aimed to verify the chemical component cantharidin from Chinese blister beetle by LC-MS/MS. Components were first processed by HPLC to choose the appropriate separation conditions before LC/MS/MS analysis. Through the positive and negative ions pre-scan, positive ions scanning way was selected in the scanning of each separate component. The scanned maps were obtained and the relative molecular mass was deduced. The results showed that 8 types of cantharidin compounds were identified, including three isomers. It was concluded that LC-MS/MS was able to determine the chemical component cantharidin from Chinese blister beetle rapidly and accurately, which was existed in the body by the form of combined amino acid.

Objective To screen for and validate unknown tumor suppressor genes (TSGs) in sporadic colorectal cancer (CRC) patients.Methods Through loss of heterozygosity (LOH) analysis on chromosome 10 in sporadic CRC,we have found D10S185 (10q23.31-24.33 ) exhibit a higher LOH frequency in our previous study.In present study,we screen for unknown TSGs in this region through the microarray.The expression of the new gene was analyzed by quantitative real-time polymerase chain reaction (qRT-PCR).RT-PCR,immunohistochemistry and Western blot were done in colorectal cancer tissues with their pair-matched normal tissues in 50 cases to validate the results of microarray.Results Through the microarray-based high throughput screening,we found 4 significant down-regulated genes:PLCE1,CPEB3,NKX2-3 and SEMA4G,among them the down-regulation of PLCE1 was most significant.The results of qRTPCR were in relative agreement with the DNA microarray data.RT-PCR,immunohistochemistry and Western blot also showed that the expression of PLCE1 was at low levels in 46％ cancer tissues compared with normal tissues,more frequent in the poor differentiation tumor in patients under age 60 years (P ＜ 0.05 ).Conclusions This study demonstrated that down-regulation of PLCE1 was related to the tumorigenesis of sporadic colorectal cancer.PLCE1 might play a suppressive role in the development of colorectal cancer.

BACKGROUND: The degradable poly (3-hydroxybutyrate-co-3-hydroxyhexanoate) (PHBHHx) has superior mechanical property and biocompatibility.OBJECTIVE: To elucidate the intravascular biocompatibility of PHBHHx in vivo.METHODS: We developed hybrid materials based on decellularized xenogenic vascular scaffolds that were coated with PHBHHx and implanted it into the abdominal aorta of New Zealand rabbits. The decellularized xenogenic pulmonary artery patch without PHBHHx coating served as the control. The implanted patches were determined for the histology, immunofluorescence staining, scanning electron microscopy and calcium contents at 1, 4 and 12 weeks after the surgery.RESULTS AND CONCLUSION: Hybrid patches exhibited smooth lumen surface without thrombus, the intimal hyperplasia was mild and recellularization was complete; immunofluorescence staining showed that the endothelial cells in the neointima were positive for CD31, with continuous single-layer arrangement, interstitial cells were positive for smooth muscle actin; the calcium content in hybrid patches was obviously lower than that in uncoated patches. PHBHHx shows a remarkable intravascular biocompatibility in vivo and is believed as an ideal candidate for lumen coating of cardiovascular tissue engineering.

Aim To study the effect of 6-(3-Benzyl-4-oxo-2-thioxo-thiazolidin-5-ylidenemethyl)-9-fluoro-3-methyl-10-(4-methyl-piperazin-1-yl)-2,3-dihydro-7-oxo-7-hydro-pyrido[1,2,3-de][1,4]benzoxazine (R3) on apoptosis of the human hepatocarcinoma SMMC-7721 cells (in vitro).Methods With different concentrations of R3 used to treat SMMC-7721 cells, esophageal squamous cell carcinoma EC-9706 cells, human colon adenocarcinoma cell line Caco-2 cells and in human L-02 hepatocytes (in vitro), and the inhibition effects of R3 on cell proliferation were examined by MTT assay.Cell apoptosis was determined using DAPI fluorescence staining and TUNEL method.The cell cycle was detected using flow cytometry with PI staining.Protein expression of p53 and caspase-3 was detected with Western blot analysis.Results Treatment with R3 (2~20 μmol·L-1) potently inhibited the proliferation of the cancer cells (the IC50 value at 24 h in SMMC-7721 cells, EC-9706cells and CaCO-2 cells was 3.893, 4.181 and 3.408 μmol·L-1, respectively).In contrast, R3 had weak cytotoxicity against L-02 cells with IC50 value of 38.96 μmol·L-1.Ofloxacin had weak cytotoxicity against SMMC-7721 cells with IC50 value of 240.137 μmol·L-1.Sunitinib had cytotoxicity against SMMC-7721 cells with IC50 value of 8.075 μmol·L-1.Treatment of SMMC-7721cells with different concentrations of R3 for 24 h increased the percentage of the apoptosis cells (P<0.05) and caused insufficient preparation for G1/S transition.In addition, R3 increased protein expression of p53, caspase-3 and the cleaved activated forms of caspase-3 in SMMC-7721 cells.Conclusion R3 as a kind of ofloxacin rhodanine derivatives exerts potent and selectively anticancer activity through the induction of apoptosis of SMMC-7721 cells.