Humans ; Liver Cirrhosis, Biliary ; diagnosis ; etiology ; therapy

Objective To investigate the relationship between in vivo anti-tumor efficacy of Shenqi injection and its effect on immunologic function in H22 tumor-bearing model mice,and explore the cellular immunologic mechanism of Shenqi injection in inhibiting tumor growth.Methods Three different doses of Shenqi injection were given to H22 tumor-bearing model mice in treatment groups.The tumor growth inhibitory rate(IR) and the index of phagocytosis of peritoneal macrophage were calculated.MTT assay was used to observe the effect of Shenqi injection on spleen lymphocytes stimulated by ConA in vitro separated from H22 tumor-bearing mice.Murine serum IL-2 and IFN-? were detected by means of ELISA assay.Results IR was significantly reduced in two treatment groups compared with that of the model mice(60.72%,48.65%,P

Objective: To investigate the inhibitory effects of PTEN gene transfection combined with L-OHP on human cholangiocarcinoma cell line, QBC939, providing a new method for gene therapy of human biliary duct carcinoma. Methods: A eukaryotic expression vector containing PTEN gene was transfected into human QBC939 cells under mediation of lipofectamine and positive cell clones were selected and amplified. Expression of PTEN gene was detected by immunohistochemistry. MTT test was used to determine the in vitro activity of cells, electron microscope was applied to observe cell ultrastructure, and flow cytometry was used for determining the cell cycle and apoptosis. In vitro test was used to study the invasive ability of cells before and after treatment. Results: After transfected with PTEN gene, QBC939 cells had a higher expression of PTEN gene (P

Objective To evaluate the influence of narrow-band ultraviolet B on lesional microvessel density (MVD),vascular endothelial growth factor (VEGF),matrix metalloproteinases 2 (MMP-2)as well as on serum VEGF in patients with psoriasis vulgaris(PV).Methods Fifteen patients with PV were recruited into this study with 10 normal human controls.All patients received NB-UVB phototherapy thrice a week for 4-5 weeks.Prior and after the treatment,psoriasis area and severity index (PASI)was calculated,tissue specimens were taken from non-photoexposed lesions,and sera samples were obtained from these patients.Then,MvD and the expression level of VEGF and MMP-2 were measured by immunohistochemical labeled dextran polymer(LDP)method in the tissue specimens.Also,the serum level of VEGF was tested by enzyme-linked immunosorbent assay(ELISA).Results PASI score remarkably decreased in patients after the photothempy(t=13.35,P＜0.01).The MVDs were 20.52±5.02,7.33±1.24 and 4.26±0.79 capillaries per high power field(400 × amplification),in psoriatic lesions before treatment,after treatment,and normal control tissues,respectively,with a significant difference among the three groups (F=97.57,P＜0.05),and a significant increase was observed in the lesions before treatment compared with those after treatment and normal controls.The serum level of VEGF was 307.55±121.65 ng/L in psoriatic lesions before treatment,significantly higher than that after treatment(163.92±95.57 ng/L),and in normal control skin (139.78±79.06 ng/L),whereas there was no significant difference between the latter two groups(P＞0.05).The positivity rate of MMP-2 was similar among the three groups without statistical difference(P＞0.05).In psoriatic patients,a positive correlation was observed among PASI score,MVD,lesional and serum VEGF levels(P＜0.05),also among the MVD,VEGF and MMP-2 levels in lesions(P＜0.05).but lesional MMP-2 was unrelated to PASI score or sgrum VEGF(both P＞0.05).Conclusions NB-UVB may regulate superficial dermal microvascular proliferation by acting on the expression of VEGF in sera and lesions of psoriatic patients.VEGF and MMP-2 may bOth participate in the proliferation process of microvessels,while MMP-2 is unlikely to be involved in the therapeutic mechanism of NB-UVB.

Objective To investigate the mechanism of soluble β-1, 3-D-glucan in G-test positive serum in inhibiting ROS-dependent killing of Candida albicans ( C.albicans ) mediated by neutrophil Dectin-1.Methods The expression and distribution of internalized Dectin-1 and triggered ROS in human neutrophils were detected by using confocal/two-photon laser scanning microscopy upon stimulation with C.albicans (MOI=10) which was pretreated with β-1, 3-D-glucanase (10 U/ml) or not.Abrogation test was used to analyze whether intracellular Dectin-1 was involved in C.albicans-triggered ROS production in human neutrophils.Furthermore, flow cytometry analysis was performed to detect the expression of intracel-lular Dectin-1 and ROS in neutrophils which were pretreated respectively with G-test positive serum at differ-ent dilutions for 60 min and then stimulated with C.albicans for another 60 min at 37℃.Results After stimulated with C.albicans (MOI=10) for 60 min, the expression of Dectin-1 in neutrophils was recruited to the spores of opsonophagocytized C.albicans, and partly co-localized with the triggered ROS production . However, the expression of intracellular Dectin-1 was not observed in neutrophils when stimulated with β-1, 3-D-glucanase pretreated C.albicans for 60 min at 37℃.Abrogation test further showed that C.albicans-trig-gered ROS production in neutrophils was partly and irreversibly inhibited by adding Dectin -1 blocking mAb of 5 μg/ml.In addition , both the triggered expression of intracellular Dectin-1 and ROS production in neu-trophils stimulated with C.albicans ( MOI=10 ) in the presence of G-test positive serum were significantly lower than those of neutrophils stimulated only with C.albicans (LSD-t test, P<0.01).Linear regression a-nalysis suggested that the triggered intracellular Dectin-1 and ROS production in neutrophils upon stimulation with C.albicans were both inhibited by soluble β-1, 3-D-glucan in a dose-dependent manner (Dectin-1,R2=0.702,P<0.01;ROS,R2=0.588,P<0.01 ).Conclusion Taken together, these results demonstrated that the soluble β-1, 3-D-glucan in G-test positive serum may play a role in inhibiting the ROS-dependent killing of C.albicans by interfering with internalized expression of neutrophil Dectin-1.

Objective To investigate the mechanism hy which Dectin-1 mediates the killing of Candida albicans(C.albicans) by human neutrophils in a manner dependent on the production of reactive oxygen species (ROS).Methods After stimulation with FITC-C.albicans at a multiplicity of infection (MOI) of 10 for 30 or 60 min,PE-anti-human Dectin-1 monoclonal antibody (2.5 μg/106 cells) was used to detect the expression of Dectin-1 in human neutrophils by flow cytometry.For Dectin-1 inhibition test and ROS assay,human neutrophils (2×106/ml) were respectively pre-incubated with different concentrations of blocking antibody (0.5,1,2.5 and 5 μg/ml) for 60 min at 4℃,and then with 25 μmol/L 2′,7′-Dichlorofluorescein diacetate for another 20 min at room temperature.Afterwards,under stimulation with live C.albicans at a MOI of 10,the rate of intracellular ROS production over time in blocking and control groups was measured continuously at 10 min intervals for up to 120 min.In addition,localization of Dectin-1 and ROS in human neutrophils was observed by confocal/two-photon laser scanning microscopy after stimulation with live C.albicans.For the detection of candicidal activity,after pre-treatment with different concentrations of Dectin-1 blocking antibody as mentioned above,neutrophils were stimulated with live C.albicans (MOI=1) for 60 min,serial dilutions of cell lysate were plated onto yeast agar,and CFU were enumerated after incubation at 37℃ for 48 h.The candicidal activity was represented as [1-(CFUblocking group/CFUbuffer)] × 100％.Results Under stimulation with FITC-C.albicans at a MOI of 10 for 30 and 60 min,positive percentage of intracellular Dectin-1-expressing neutrophils increased significantly when compared with initial level (0 min,8.32％ ; 30 min,16.82％ ; 60 min,23.88％) (versus 0 min,P＜0.01).However,positive percentage of cell-surface Dectin-1-expressing neutrophils remained almost unchanged after stimulation for 30 and 60 min (versus 0 min,P＞0.05).Upon blockage of Dectin-1,the stimulated ROS generation (R2=0.306,P＜0.01) and candicidal activity (R2=0.251,P＜0.01) of neutrophils were partly and irreversibly inhibited in a dose-dependent manner when compared with control group.In addition,the intracellular Dectin-1 is recruited and co-distributed with ROS on the surface of phagocytized C.albicans as observed by confocal microscopy.Conclusion Taken together,these results demonstrated that an internalized expression pattern of human Dectin-1 might contribute to the ROS-dependent killing of serum-opsonzied C.albicans which was phagocytized by human neutrophils.

NanoString nCounter Analysis System is a newly developed gene expression detection platform that directly measures multiplexed mRNA levels through digital counting of individual mRNA transcripts.This technology uses as little as 100 ng of RNA and can obtain accurate gene quantitative data from up to 800 genes in one reaction.It requires no reverse transcription,enzymes and amplifications,and its sensitivity and accuracy are comparable to real time quantitative PCR.NanoString technology has been more and more extensively used in frontiers of biomedical research and clinics such as in validation of data from high-throughput platforms,gene expression profiling,gene regulatory network,molecular subtyping,diagnosis and prognosis of diseases.

BACKGROUND:Currently, there is no effective treatment for keloids that often recur. Its pathogenesis is stil entirely unclear, and fibroblast proliferation and apoptosis have become a research hotspot.
OBJECTIVE:To investigate the expression of Livin, Smac and Caspase-3 in keloids and to analyze their relationship so as to preliminarily explore the significance of Livin, Smac and Caspase-3 in the pathogenesis of keloids.
METHODS:RT-PCR and immunohistochemical methods were used to detect the mRNA and protein expressions of Livin, Smac and Caspase-3 in keloids (n=20) and normal skin tissues (n=20).
RESULTS AND CONCLUSION:Compared with the normal skin tissue, the mRNA and protein positive expressions of Livin were significantly higher in keloids (P < 0.05), while the mRNA and protein positive expressions of Smac and Caspase-3 were lower in keloids (P < 0.05). There was a negative association between Livin and Smac, Caspase-3 protein expression in keloids. These findings indicate that the high mRNA expression of Livin may cause the imbalance between proliferation and apoptosis of fibroblasts by inhibiting the mRNA expression of Smac and Caspase-3, and eventualy lead to the formation of keloid.

Adult ; Animals ; Colitis ; chemically induced ; pathology ; Colon ; pathology ; Drugs, Chinese Herbal ; administration & dosage ; adverse effects ; Female ; Humans ; Indigo Carmine ; Indoles ; administration & dosage ; adverse effects ; Male ; Middle Aged ; Phytotherapy ; Psoriasis ; drug therapy ; Rats ; Rats, Sprague-Dawley

50%.The patients were categorized into as:one-,two-, and three-vessels coronary artery disease group.Central aortic SBP and DBP was measured by cathetarization dur- ing angiography of coronary artery and brachial blood pressure was measured using cuff method.Results Periph- eral SBP,PP and ascending aortic SBP,PP,fractional systolic pressure(FSP=SBP/MAP)were increased and as cending aortic fractional diastolic pressure(FDP=DBP/MAP)was reduced when the diseased coronary vessels were increased(P