Objective To investigate the expressions of stern cell markers CD133,nestin and CD44 in malignant melanoma and their significance.Methods Tissue samples were obtained from 30 patients with malignant melanoma and 30 patients with intradermal nevus.The expressions of three markers were immunohistochemically detected in the samples.Results In malignant melanoma specimens,the expression rate of CD133,nestin and CD44 was 53.33%(16/30),80.00%(24/30)and 20.00%(6/30),respectively,significantly difierent from that in intradermal nevus specimens [23.33%(7/30),53.33%(1 6/30)and 0,respectively,all P＜0.05].The percentage of cells positive for CD133,nestin and CD44 was 2.98%±5.62%,34.92%±34.89%and 1.28%±3.26%,respectively,in malignant melanoma specimens.0.10%±0.21%,7.26%±13.13%and 0,respectively,in intradermal nevus specimens;there was a significant difierence between the two groups of specimens(all P＜0.05).In malignant melanoma and intradermal nevus,the expression intensity of CD133.nestin and CD44 showed no significant correlation with patients'sex.age or disease course(all P＞0.05).ConclusionsCD133,nestin and CD44 are highly expressed in malignant melanoma,but weakly expressed or absent in intradermal nevus,suggesting that tumor stem cells might exist in malignant melanoma tissue.

Objective:To investigate the function of CD 59 in LAT induced T lymphocytes'proliferation and activation.Methods:Transfected LAT-GFP recombinant lentiviral vectors into Jurkat cells and established a fusion-protein stable express cell line ( Jurkat-GFP ).Junket-GFP cells were transfected with pSUPER-siCD59 plasmids by electroporetion or stimulated by anti-CD59 antibody.The cellular locations of CD 59 and LAT were observed under fluorescence microscope with the immunofluorescence cytochem -istry.The cells proliferation were measured by MTT assay.Furthermore,Western blot was used to detect the total and phosphorylation levels of several down-stream proteins after T cell activated .Results: Jurkat-GFP cells successfully transfected with pSUPER-siCD59 plasmids showed lower fluorescence staining.CD59 and LAT distributed uniformly on the cell surface before stimulated with anti-CD59 antibody and formed clusters once upon stimulation.Jurkat-GFP cells stimulated with anti-CD59 antibody showed a higher level of pro-liferation and protein phosphorylation ,compared with the others.Conclusion:CD59 contributed to LAT induced signaling transduction of T lymphocytes ,and stimulated CD59 molecule partly promoted T cell activation.

ObjectiveTo establish a method for content determination of total iridoid compounds and baldrinal and 11-ethoxyviburtinal from Valerianae Jatamansi Rhizoma et Radix; To determine the contents of total iridoid compounds and baldrinal and 11-ethoxyviburtinal in Valerianae Jatamansi Rhizoma et Radix from three medicinal origins.Methods UV spectrophotometry was applied, 11-ethoxyviburtinal (cyclopentane-pyran-7-formaldehyde, 4-ethoxy methyl) was set as the reference substance, and the content of total iridoid compounds was determined at 288 nm. HPLC method was used to simultaneously determine the contents of baldrinal and 11-ethoxyviburtinal. The HPLC analysis was performed on a Phenomenex Luna C18 column (250 mm×4.6 mm, 5μm). The mobile phase was composed of acetonitrile-water in gradient elution at a flow rate of 0.95 mL/min. The detection wavelength was 288 nm and the column temperature was 30℃.Results The total iridoid compounds, baldrinal and 11-ethoxyviburtinal were in good linearity within the ranges of 2.088–14.616μg/μL, 74.88–224.64μg, and 41.6–249.6μg, respectively. This method was precise, and with good repeatability, stability and recovery rate.Conclusion The method is accurate, simple, rapid, which can be used for the quality control of Valerianae Jatamansi Rhizoma et Radix.

Objective:To investigate the effects of chitin on atopic dermatitis in an OVA induced AD murine model.Methods:Twenty-eight BALB/c mice were randomly divided into three groups:the normal control group (N)(8),the chitin group(E) (10) and the AD model group(M)(10).The murine model of atopic dermatitis was established through intraperitoneal injection of OVA followed by repeated epicutaneous application of OVA on mice back skin( AD model group).During the set up of AD murine model,mice of the chitin group were given intragastric gavage of 3 mg/d for 4 weeks.At the end of the experiment, the mice were sacrificed and skin lesions were biopsied for histological study.HE and O-toluidine stained paraffin sections were observed under microscope.The spleen cells were cultured and challenged with OVA and chitin,respectively,the supernatant was obtained for cytokine determination.Serum levels of total and OVA-specific IgE and total IgG2a were determined with ELISA.Results:Chitin significantly inhibited skin inflammation induced by OVA.Compared with the AD model group,the thickness of the epidermis and dermis in the chitin group were obviously decreased.The numbers of dermal infiltrated inflammatory cells,eosinophils and mast cells were significantly decreased in the chitin group compared with the AD model group ( P<0.05-0.001 ).The serum level of total IgE and OVA-specific IgE were significantly lower in the chitin group than in the AD model group(P<0.05-0.001),while the serum level of IgG2a in the chitin group was significantly higher than that of the AD model group( P<0.001).The cultured spleen cells of the chitin group produced significantly higher levels of IL-12 and IFN-γ,but lower level of IL-4 compared with those of the AD model group after OVA challenge (P<0.05).Conclusion:Chitin can inhibit the inflammation and decease the seum level of IgE in the murine AD model.The antiallergic effect of chitin might be associated with the induced production of Th1 type cytokines by mice spleen cells.

BACKGROUND:The linkage and synergistic effect of adaptor proteins can effectively regulate signal transduction of T cel s, which can form a limit or amplification cascade to realize the complex immune function of T cel s. C-terminal Src kinase (Csk)-binding protein (Cbp) is an adaptor protein, which mainly exert the negative feedback regulation of Src kinase activity. This negative feedback effect depends on Y317 of Cbp, which may be involved in the SH2 domain of Csk. OBJECTIVE:To explore the effects of high expression of Cbp on ultrastructure and related biological function of Jurkat cel s. METHODS:The virus particles were constructed with expressing enhanced green fluorescent protein (EGFP) only and Cbp-EGFP fusion protein to transfect Jurkat cel s. There were untransfected group (Jurkat group), negative control group (transfected with expression of EGFP virus only), and Cbp group (transfected with Cbp-EGFP virus). RESULTS AND CONCLUSION:Confocal microscope showed that cel transfection efficiency was more than 95%and Cbp was located on the cel membrane. Optical microscope showed after transfection with Cbp-EGFP virus, more Jurkat cel s shrunk, with poor size uniformity. Apoptosis detection showed that after transfection with Cbp-EGFP virus, the number of apoptotic and necrotic cel s was greatly increased. Cbp mRNA expression was increased, Csk expression was decreased obviously and lymphocyte-specific protein tyrosine kinase expression was increased. So, in Jurkat cel s, the high expression of Cbp can decrease the uniformity of cel s and increase the necrosis cel s, thus inhibiting the signal transduction.

In order to solve the problem of selection and in vivo delivery problem in siRNA treatment, hepatitis B virus (HBV) HBx gene which could be targeted by siRNA was studied. The siRNA expression plasmid which specific inhibits HBx expression was obtained by in vitro selection via a dual-luciferase plasmid including HBx-Fluc fusion protein expression domain. The selected siRNA expression plasmid was then encapsulated in PEG-modified cationic liposome, which was devoted into pharmacodynamic studies at both cellular and animal level. The results illustrated that the cationic liposome which encapsulated siRNA expression plasmid could effectively inhibit HBx gene expression both in vitro and in vivo.

Objective To investigate the immunophenotype and molecular genetics of mature aggressive B-cell lymphomas in Chinese pediatric patients and provide the criteris for the diagnosis of them.Methods We collected 97 paraffin-embeded tissue samples of pediatric cases of mature aggressive B-cell lymphomas including 81 Burkitt lymphoma (BL) cases, 8 diffuse large B cell lymphoma (DLBCL) cases and 8unclassifiable B cell lymphoma with featares intermediate between BL and DLBCL (BL/DLBCL) cases. The immunophenotype and genetic features of them were detected by immunohistochemistry and interphase FISH.Results The expression of bcl-2 [3 %(2/66) in BL, 50 % (4/8) in DLBCL, 50 % (4/8) in BL/DLBCL], MUM1 [17 % (12/71) in BL, 63 % (5/8) in DLBCL, 63 % (5/8) in BL/DLBCL] and mean Ki-67 proliferation index [(93±4.4)% in BL, (83±14.3)% in DLBCL, (80±11.5)% in BL/DLBCL] were significantly different between BL and DLBCL and between BL and BL/DLBCL. The frequency of c-myc rearrangement [98 % (79/81) in BL,38 % (3/8) in DLBCL, 50 % (4/8) in BL/DLBCL] and an extra copy of bcl-6 [0 % in BL, 38 % (3/8) in DLBCL, 25 % (2/8) in BL/DLBCL] were also significantly different between BL and DLBCL and between BL and BL/DLBCL. Conclusion Diagnosis of the mature aggressive B cell lymphomas in pediatrics should be based on the comprehensive review and integration of morphologic, immunohistochemical and molecular genetic features. BL/DLBCL is more likely a subgroup of the DLBCL in pediatric population. The expression of CD10 and bcl-6 but not bcl-2, a high Ki-67 PI (＞90 %) and a c-myc rearrangement but not bcl-2 or bcl-6rearrangement are the features of BL. Regardless of the expression of CD10 and bcl-6, positive staining for bcl2, Ki-67 PI below 90 % and an extra copy of the bcl-6 favor a diagnosis of DLBCL or BL/DLBCL.

Objective To compare the changes of serum C reactive protein (CRP) in different lesion site and activity so as to evaluate its worthy of an indicator of disease activity. Methods Forty-two patients with Crohn's disease (CD) were divided into small intestinal group and colonic group according to the involved lesions. Twenty-three cases of UC and 26 cases of inflammatory bowel disease (IBS)were served as controls. The serum level of hs-CRP was tested using latex-enhanced immunoturbidimetery. mg/L and (1.1±1.8)mg/L, respectively. Hs-CRP was elevated significantly in CD group compared to UC and IBS groups (P＜0.001). The ratio of patients whose hs-CRP exceeded 3 mg/L was 76.2%, 30.4% and 7.7% in CD, UC and IBS, respectively (P=0.000). The ratio was significantly higher in CD higher than that of small intestinal group [(11.9±7.6 )mg/L vs (6.8±7.2)rag/L, P =0.04]. The ratio of patients whose hs-CRP exceeded normal value was higher in colonic group than that in small CRP(≥10 rag/L). Among them, 4/17 were in remission, 3/11 in mild, 10/13 in moderate and 1/1 in severe according to the CDAI. The hs-CRP was correlated well with CDAI and ESR (r was 0.52 and 0.70 respectively, P＜0.001). Conclusions CRP can he used as a inflammatory marker for evaluating the disease activity of CD. The patients with small intestinal involvment may have lower CRP than those with colonic affection. The elevation of CRP was paralleled to the disease severity of CD.

Apoptosis ; Cell Proliferation ; Humans ; Neoplasms ; Tumor Suppressor Protein p53

Objective To investigate the levels of cancer related fatigue (CRF) and the correlations between CRF and serum inflammatory factors and hypothalamus-pituitary-adrenal (HPA) axis in patients with gastrointestinal cancer.Methods The CRF level was assessed by brief fatigue inventory (BFI).The level of C-reactive protein (CRP) was measured by immunoturbidimetry, and the level of cortisol was measured by electrochemiluminesence.The levels of interleukin (IL)-6, IL-1β, tumor necrosis factor-α (TNF-α), adrenocorticotropic hormone (ACTH) and norepinephrine (NE) were measured by enzyme-linked immunosorbent assay (ELISA).Results The average total score of CRF was 3.15±1.93, and the degree was mild to moderate, which was positively correlated with the CRP (r=0.321, P=0.000), TNF-α (r=0.265, P=0.000), NE (r=0.174, P=0.015) and ACTH (r=0.257, P=0.000), but was not correlated with the cortisol (r=0.033, P=0.652).Eastern Cooperative Oncology Group (ECOG) score (t=8.081, P=0.000), education (t=-4.244, P=0.000), treatment (t=4.563, P=0.000), time from diagnosis to sampling (t=3.453, P=0.001) and CRP (t=2.837, P=0.006) were important factors of CRF.Conclusion The CRF status is common in gastrointestinal cancer patients.The CRF is correlated with the NE and ACTH of HPA axis.Medical staff should pay attention to the inflammatory factors and hormone levels to improve the fatigue status and the quality of patients.