ObjectiveTo explore whether the biocompatibility of phosphorylcholine (PC) modified alginate-chitosan microcapsules could be improved. MethodsPC modified alginate-chitosan microcapsules were obtained by high-voltage electrostatic system.Bradford method was adopted to determine the adsorption amounts of bovine serum albumin by chitosan alone and PC modified chitosan.Alginate-chitosan-PC microcapsules (experimental group) and alginate-chitosan microcapsules ( control group) were respectively implanted into the peritoneal cavity of mice and retrieved 4 weeks after transplantation.Fibrosis of the capsules was evaluated by HE staining.Glucose stimulated insulin secretion (GSIS) assay was used to assess the insulin secretion response of encapsulated and nonencapsulated rat islets. Results The adsorption amount of protein was 189.4 μg/mg and 90.5 μg/mg respectively by chitosan alone and PC modified chitosan.The difference had statistical significance ( t =5.549, P ＜ 0.05 ).In contrast to the control group, the cellular reaction on the surface of the modified microcapsules was weaker, with no obvious fibrosis found.The insulin secreted by encapsulated islets and nonencapsulated islets was( 3.298 ± 1.680 ) μIU/ml VS (4.299 ± 1.159 ) μIU/ml ( t =1.096, P ＞ 0.05 ) in response to low-glucose stimulus and( 11.783 ± 4.175 ) μIU/ml VS ( 12.875 ± 2.268 ) μIU/ml ( t =0.514, P ＞ 0.05 ) in response to high-glucose stimulus.Conclusions PC can improve the biocompatibility of alginate-chitosan microcapsules, with no effect on the biological function of encapsulated islets.It may be more appropriate to use modified microcapsules encapsulating islets for transplantation.

Carcinoma ; genetics ; Humans ; In Situ Hybridization, Fluorescence ; methods ; Male ; Pancreatic Neoplasms ; genetics ; Spectral Karyotyping ; methods

OBJECTIVETo investigate whether the serum levels of inflammation-related cytokines might be different between the healthy individuals and the psoriatic patients diagnosed of three varied Chinese medicine (CM) syndromes [blood-stasis syndrome (BSS), blood-dryness syndrome (BDS) and wind-heat syndrome (WHS)].

METHODSA total of 62 psoriatic patients were recruited and assigned to 3 groups according to their CM syndromes, including 27 patients of BSS, 21 of BDS and 14 of WHS. Another 20 sex- and age-matched healthy subjects were enrolled into the control group. Serum concentrations of multiple cytokines, including monocyte chemotactic protein-1 (MCP-1), macrophage inflammatory protein-1α (MIP-1α), soluble CD4O ligand (SCD40L), tumor necrosis factor-α (TNF-α), epidermal growth factor (EGF), interleukin-8 (IL-8), interleukin-17 (IL-17), interferon γ inducible protein-10 (IP-10) and vascular endothelial growth factor (VEGF), were measured by a multiplexed flow cytometric assay.

RESULTSThe circulating levels of MIP-1α, TNF-α, IL-8, and IP-10 were significantly increased in the psoriatic patients compared with the healthy controls (P<0.01). Male and female patients tended to have higher serum levels of MCP-1 and IP-10, respectively (P<0.05). Interestingly, compared with the control group, 6 out of the 9 cytokines (MCP-1, MIP-1α, TNF-α, EGF, IL-8 and IP-10) were substantially increased in the BSS group (P<0.05 or P<0.01), whereas only MIP-1α and IL-8 levels were elevated in the BDS group (P<0.05 or P<0.01) concurrent with lowered concentrations of SCD40L and IL-17 (P<0.05). In the WHS group, MIP-1α was the only cytokine whose level was evidently increased (P<0.01), in contrast to IL-17 which was decreased as compared with the control (P<0.05). The psoriatic patients overall owned higher levels of MIP-1α and IL-8 in the circulation which were comparable among the 3 groups of CM syndromes (P<0.01). In contrast, TNF-α level of the BSS group was the highest among the three (P<0.01), followed by the BDS and the WHS groups.

CONCLUSIONSThe expression profiles of cytokines in the circulation might not be necessarily identical for psoriatic patients with different CM syndromes. Accordingly, the serum concentrations of certain cytokines could potentially be used as the ancillary indices for the clinical classification of psoriatic CM syndromes.

Adult ; Aged ; Case-Control Studies ; Cytokines ; blood ; Female ; Humans ; Inflammation Mediators ; blood ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Psoriasis ; blood ; Severity of Illness Index ; Statistics, Nonparametric ; Syndrome ; Young Adult

In recent years,the CTLA-4 immunoglobulin biologics,a negative regulator in the immune system,have been obtained due attention in autoimmune diseases,transplantation rejection,and antineoplastic agents.CTLA-4 can inhibit T cell activation,reduce the expression of RANKL and other cytokines through regulating immune response,and effectively alleviate the process of bone resorption.According to previous study,CTLA-4 was involved in osteoclast-induced bone destruction and bone remodeling.In this review,the effect of CTLA-4 on the autoimmune diseases,on the osteoclast formation,and on the alveolar bone remodeling in the periodontal tissue was involved,and the related research were also evaluated to look forward to possible future basic research and clinical application direction.

Objective:To analyze therapeutic effect of amiodarone combined RAS inhibitors on hypertensive patients with atrial fibrillation(AF),and its influence on serum levels of uric acid(SUA)and silent information regulator 2-related enzymes 1(SIRT1).Methods:A total of 186 hypertensive patients with AF were selected from our hospital. They were randomly and equally divided into amiodarone group(group A),amiodarone + telmisartan group(A+T group)and amiodarone + ramipril group(A+ R group).Left atrial diameter(LAD),P wave dispersion(Pd),lev-els of high sensitive C reactive protein(hsCRP),brain natriuretic peptide(BNP)and SUA,SIRT1 mRNA and pro-tein expression and blood pressure variability(BPV)were observed and compared among three groups before and 45d after treatment.Results:Compared with before treatment,there were significant improvement in all index ex-cept SUA after treatment in three groups,P=0.001 all;compared with group A after treatment,there were signif-icant reductions in 24hSBPV[(0.112 ± 0.022)vs.(0.092 ± 0.020)vs.(0.091 ± 0.021)],24hDBPV[(0.143 ±0.031)vs.(0.130 ± 0.040)vs.(0.129 ± 0.039)],levels of hsCRP[(8.3 ± 0.3)mg/L vs.(5.3 ± 0.6)mg/L vs. (3.6 ± 0.4)mg/L],BNP[(64.9 ± 9.4)ng/dl vs.(61.1 ± 7.6)ng/dl vs.(58.2 ± 8.3)ng/dl]and SUA[(498.85 ± 89.54)μmol/L vs.(298.54 ± 56.12)μmol/L vs.(278.32 ± 54.09)μmol/L],SIRT1 mRNA[(2.20 ± 0.34)%vs.(1.87 ± 0.30)% vs.(1.76 ± 0.31)%]and protein[(29.54 ± 8.12)% vs.(26.31 ± 7.43)% vs.(23.21 ± 6.90)%]expression in A+T group and A+ R group,P<0.05 or <0.01. Compared with group A,there were sig-nificant rise in Pd[(39.3 ± 4.2)ms vs.(40.9 ± 4.1)ms,(41.2 ± 5.1)ms],and significant reduction in LAD [(37.8 ± 3.4)mm vs.(35.1 ± 4.6)mm,(35.7 ± 4.5)mm]in A+T group and A+R group,P<0.05 or <0.01. Conclusion:Amiodarone combined RAS inhibitors can significantly improve blood pressure variability,and reduce inflammatory factor,SUA and serum SIRT1 level in hypertensive patients with AF.

OBJECTIVEto investigate the chromosomal characteristics of pancreatic ductal adenocarcinomas by spectral karyotyping.

METHODScytogenetic aberrations of pancreatic cancer cell line P2 established from a Chinese patient was investigated by spectral karyotyping (SKY). Chromosomal alterations were further evaluated in 10 cases of pancreatic cancer and 10 cases of chronic pancreatitis by two color fluorescence in situ hybridization (FISH) by using EGFR/CEP7 probe and paraffin embedded tissue samples.

RESULTShypotriploid and 26 chromosomal aberrations were revealed in cell line P2. Recurrent chromosomal numerical alterations included loss of chromosome 4, 9, 18, 19, 22, Y, 10p, 15p, 8p, 6q and 12p, with gain of chromosome 7 and 12q. Frequent chromosomal structural abnormalities included der(9;15)(q10;q10), der(10)(3;10)(?;q26) and der(12)(8;12)(?;p13). Four of 10 cases showed EGFR copy number changes by FISH.

CONCLUSIONShighly complex chromosomal rearrangements occur in pancreatic cancers. Additional studies of more cases are needed, including FISH analysis of EGFR copy number changes, to reach a conclusion.

Adenocarcinoma ; genetics ; Aged ; Cell Line, Tumor ; Chromosome Aberrations ; Chromosome Deletion ; Chromosome Duplication ; Female ; Gene Dosage ; Genes, erbB-1 ; genetics ; Humans ; In Situ Hybridization, Fluorescence ; Karyotyping ; methods ; Male ; Middle Aged ; Pancreatic Neoplasms ; genetics

The apoptotic effects of shikonin (5,8-dihydroxy-2-[(1R)-1-hydroxy-4-methylpent-3-enyl]naphthalene-1,4-dione) on the human colon cancer cell line SNU-407 were investigated in this study. Shikonin showed dose-dependent cytotoxic activity against SNU-407 cells, with an estimated IC50 value of 3 µM after 48 h of treatment. Shikonin induced apoptosis, as evidenced by apoptotic body formation, sub-G1 phase cells, and DNA fragmentation. Shikonin induced apoptotic cell death by activating mitogen-activated protein kinase family members, and the apoptotic process was mediated by the activation of endoplasmic reticulum (ER) stress, leading to activation of the PERK/elF2α/CHOP apoptotic pathway, and mitochondrial Ca2+ accumulation. Shikonin increased mitochondrial membrane depolarization and altered the levels of apoptosis-related proteins, with a decrease in B cell lymphoma (Bcl)-2 and an increase in Bcl-2-associated X protein, and subsequently, increased expression of cleaved forms of caspase-9 and -3. Taken together, we suggest that these mechanisms, including MAPK signaling and the ER-and mitochondria-mediated pathways, may underlie shikonin-induced apoptosis related to its anticancer effect.
Apoptosis ; bcl-2-Associated X Protein ; Caspase 9 ; Cell Death* ; Cell Line ; Colon* ; Colonic Neoplasms* ; DNA Fragmentation ; Endoplasmic Reticulum* ; Extracellular Vesicles ; Humans* ; Inhibitory Concentration 50 ; Lymphoma, B-Cell ; Mitochondria ; Mitochondrial Membranes ; Protein Kinases

Objective To explore whether the expression level of heparanase (HPA) and its coagulation proteins on leukemic blast membrane could determine the hemostatic balance on the surface of leukemia cells.Methods Forty patients of leukemia were studied,and 20 patients with iron dificient anemia as the control group.Expression of tissue factor (TF),heparanase (HPA),tissue factor pathway inhibitor (TFPI),and urokinase plasminogen activator receptor (UPAR) on leukemic blast surfaces were analyzed by flowcytometry.Results The expression of TF,UPAR,and HPA in AML,ALL,CML,CLL and CRAL groups were significantly higher compared with the control group (t =.3.289,3.507,2.701,P ＜0.05; t =2.498,0.802,3.090,P ＜0.05; t =2.642,3.308,2.696,P ＜0.05; t =3.417,3.434,2.382,P ＜0.05; t =2.193,2.272,2.263,P ＜0.05).There were no significantly differences between the leukemic cell expression of TFPI and the control group (P ＞0.05).Expression of TF,UPAR,HPA in AML patients were significantly higher than ALL,CML and CLL groups (t =2.463,2.179,2.276,P ＜0.05; t =2.637,2.402,2.095,P ＜0.05; t =2.548,2.425,2.412,P ＜0.05).The levels of TF,UPAR and HPA in M3,M4 and M5 patients were higher than that of M1,M2 groups (P ＜0.05).There were no significantly differences among M3,M4 and M5 (P ＞0.05).Conclusions These results suggest that TF,UPAR and HPA are predominately expressed on leukemic blast surface,particularly in M3and M4,5 subtypes.The expression of coagulation proteins on blast membrane might determine the hemostatic balance on the surface of leukemia cells.

This study was aimed to investigate the frequency of FMS-like tyrosine kinase 3 (FLT3) mutations including internal tandem duplication (ITD) mutation of juxtamembrane region and point mutation of the second tyrosine kinase domain (TKD) in acute myeloid leukemia (AML) patients and its clinical significance. The ITD mutation in FLT3 exon 14, 15 of bone marrow mononuclear cells was detected by genomic DNA-PCR, the TKD point mutation in FLT3 exon 20 was detected by genomic DNA-PCR combined with restriction endonuclease digest. The results indicated that among 131 newly diagnosed AML patients, 21 patients (16.0%) showed FLT3-ITD positive, 3 patients (2.3%) showed FLT3-TKD positive. None was found harboring both mutations. The WBC and bone marrow blast counts in FLT3-ITD positive patients seemed both higher than those in patients with wild-type FLT3 (FLT3-wt), but there was significant difference only in WBC count (p<0.05). The complete remission (CR) rate in FLT3-ITD positive patients was 47.6%, which was significantly lower than that in FLT3-wt patients (88.1%, p<0.05). There was no statistical difference in CR rate between FLT3-ITD positive and negative patients in 20 cases of M3; the CR rate in FLT3-ITD positive patients with non M(3) was 37.5 (6/16) which was obviously lower than that in FLT3-wt patients with non M3 (90.6%, 48/53) (p<0.05). 3 FLT3-ITD positive patients with CR relapsed after CR for 14 (2-20) months with relapse rate 50% (3/6) which was higher than that in FLT3-wt patients (29.2%, 14/48). It is concluded that FLT3 mutation is common in AML patients, while FLT3-ITD mutation is more frequent than FLT3-TKD mutation. The AML patients with FLT3-ITD mutation have a poor prognosis, while FLT3-TKD point mutation does not significantly influences prognosis of the patients. Therefore early detection of FLT3 mutation may be important for targeting therapy and evaluating clinical prognosis of AML patients.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Child, Preschool ; Female ; Humans ; Leukemia, Myeloid, Acute ; genetics ; Male ; Middle Aged ; Mutation ; Protein Structure, Tertiary ; Young Adult ; fms-Like Tyrosine Kinase 3 ; genetics

OBJECTIVESTo determine the effect of cis-9, trans-11-conjugated linoleic acid on the cell cycle of mammary cancer cells (MCF-7) and its possible mechanism of inhibition cancer growth.

METHODSUsing cell culture and immunocytochemical techniques, we examined the cell growth, DNA synthesis, expression of PCNA, cyclin A, B(1), D(1), p16(ink4a) and p21(cip/wafl) of MCF-7 cells which were treated with various c9, t11-CLA concentrations (25 mM, 50 mM, 100 mM and 200 mM) of c9, t11-CLA for 24 and 48 h, with negative controls (0.1% ethanol).

RESULTSThe cell growth and DNA synthesis of MCF-7 cells were inhibited by c9, t11-CLA. MCF-7 cells, after treatment with various c9, t11-CLA doses mentioned above for 8 days, the inhibition frequency was 27.18%, 35.43%, 91.05%, and 92.86%, respectively and the inhibitory effect of c9, t11-CLA on DNA synthesis (except for 25 mM, 24 h) incorporated significantly less(3)H-TdR than did the negative control (P<0.05 andP<0.01). To further investigate the influence on the cell cycle progression, we found that c9, t11-CLA may arrest the cell cycle of MCF-7 cells. Immunocytochemical staining demonstrated that MCF-7 cells preincubated in media supplemented with different c9, t11-CLA concentrations at various times significantly decreased the expressions of PCNA, and Cyclin, A, B(1), D(1) compared with the negative controls (P<0.01), whereas the expressions of p16(ink4a) and p21(cip/wafl), cyclin-dependent kinases inhibitors (CDKI), were increased.

CONCLUSIONSThe cell growth and proliferation of MCF-7 cells is inhibited by c9, t11-CLA by blocking the cell cycle, which reduces expressions of cyclin A, B(1), D(1) and enhances expressions of CDKI (p16(ink4a) and p21(cip/wafl)).