The effects of Guizhi Fuling capsule and its active ingredient combination within different concentration on SPL proliferate were observed by MTT method. The ratio of CD80/86, CD3CD25 and CD3CD69 was used to evaluate cell activation effects of Guizhi Fuling capsule and its active ingredient combination by FCM. Guizhi Fuling capsule with concentration of 400 mg · L(-1)can promote spleen lymphocyte proliferation, as well as the active ingredient combination, which showed the obvious dose-effect relationship. Compared with control group, the difference has statistical significance (P≤0.01). The result of FCM showed that Guizhi Fuling capsule and its active ingredient combination can promote CD80 and CD86 expression on spleen lymphocyte, and also can increase CD25 and CD69 ratio between spleen CD3+ cells. Compared with control group, the difference has statistical significance (P≤0.01). Thus, Guizhi Fuling capsule and its active ingredient combination may have immune-modulate effects, and the mechanism may have a close relationship with the lymphocyte activation.
Animals ; Capsules ; Drugs, Chinese Herbal ; analysis ; pharmacology ; Immunologic Factors ; pharmacology ; Lymphocyte Activation ; drug effects ; Male ; Mice ; Mice, Inbred BALB C ; T-Lymphocytes ; drug effects ; immunology

Objective To observe the damage effect of borneol on rabbit corneal epithelial cells. Methods After the treatment with borneol at 100, 200, 400 μg·mL-1 respectively, the viability of rabbit corneal epithelial cells was determined by methyl thiazolyl tetrazolium ( MTT) assay, cell apoptosis was determined by flow cytometry with Annexin V- fluorescein isothiocyanate/propidium iodide ( FITC/PI) staining, and Caspase-3 mRNA expression was detected with real-time reverse transcription polymerase chain reaction ( RT-PCR) . Results Borneol at the concentrations of 100, 200, 400 μg·mL-1 inhibited the activity of rabbit corneal epithelial cells. Compared with the normal control group, borneol increased the rate of apoptosis, and enhanced the Caspase-3 mRNA expression in rabbit corneal epithelial cells ( P<0.05 or P<0.01) . Conclusion Borneol can inhibit the proliferation of rabbit corneal epithelial cells, induce cell apoptosis through enhancing the expression of apoptosis-related gene Caspase-3 mRNA.

Objective To observe the effect of low dosage of uhrashortwave(USW) on infarction volume, B cell lymphocytoma-xl (Bcl-xl) and tumor necrosis factor alpha (TNF-?) after cerebral ischemia-reperfusion in rats and discuss its acting mechanisms. Methods Focal ischemia-reperfusion model was established in 25 rats by re- versible right middle cerebral artery occlusion with filament. The right side cerebral ischemia was lasted for 2 hours and then followed with 24 hours of reperfusion. The content of neurological deficits were evaluated by the Zea-Longa 5-degree scoring system to select rats. After surgery, the rats were divided into 3 groups: blank control group, control group and USW treatment group. The brain of all rats was taken at 24 hours after reperfusion. The cerebral infarction volume, the expression of Bcl-xl and TNF-?were measured and analyzed. Results Twenty-five rats were used in the analysis of results. When compared with the control group, the infarction volume and rate in total cerebral volume of USW group significantly decreased (t = 2.54, 2.33, P

Objective To investigate the effects of tetramine to cardiac/skeletal muscle in the rats and elucidate the relationship of the effects and the elevated sero-enzyme.Method The 30 homogeneous SD rats were invided into three groups:the control group,the half-lethal dose group and the lethal dose group.The number of female and male rats was equal.The tetramine powder is dissolved into 0.9% NS and puured into the stomach of the objects in medication groups;0.9%NS was poured into the rats of control group.Once the rat died or one hours later,the related sero-enzyme was determined and the rat was executed,Immediately cardiac muscle and skeletal musclewas respectively drawn the materials from the rat and was pathologically examined.Results After the rats are intragastric administrated they spasm in 10-60 min;the 6 rats of the medial lethal dose group die in 20-60 min.Serum TnI/CK/AST/LDH/a-HBDH in medication groups is higher than in control group (P

Objective To study the effects of angiotensin converting enzyme inhibitor benazepri1 on apoptosis and the expression of Fas and FasL in the kidney of rats with adriamycin-indued nephritic glomeruosclerosis.Methods After uninephrectomy and the injection of adriamycin induced rats model with glomerulosclerosis, benazapril(6 mg/kg) was delivered daily by gavage to the rats in therapeutic groups for 12 weeks.Apoptosis was examined by means of terminal-deoxynucleotidyl trans ferase mediated d-UTP nick end label ling(TUNEL) and immunohistochemistry was utlized to detect the expression of Fas and FasL.Software of pathological analysis quantitated the level of Fas and FasL.Results Compared with those of the control group, the kidney of model group had moresevere glomerulosclerosis, much more apoptotic cells and higher level of exprssion of Fas and FasL. The degree of glomeruloscleroais, the nuxner of apoptotic cells and the level of expression of Fas and FasL were ameliofated by benazepril treatment.Conclusion Benazepril may suppress the excessive apoptosis of kidney cell by lowering the expression of the protin correlatng apoptosis Fas and FasL,so as to postpone the process of glomeruosclerosis.

NP, P and L gene of Newcastle disease virus of goose origin were amplified and cloned into pGEM-T easy vector and then subcloned into pCI-neo expression vector respectively, the positive clones were identified by enzyme cutting, PCR and sequencing. GFP reporter gene was inserted into the downstream of recombinant expression plasmid of P gene, which of stop codon was deleted. The experiment of transfection of P and GFP recombinant plasmid on COS-1 cells and CEF showed that GFP gene expressed, and this demonstrated that P gene was also expressed. This research may be helpful for further study of reverse genetics and functional genome of NDV of goose origin.

OBJECTIVETo investigate the apoptosis effect of diffuse large B-cell lymphoma cell line (DLBCL) SUDHL-4 induced by IL-21 and its related mechanism.

METHODSSUDHL-4 cells were treated with IL-21 at different concentration (1000 ng/ml, 100 ng/ml, 10 ng/ml, 1 ng/ml) for 24 h, 48 h, 72 h, respectively. The inhibitory rate of cell proliferation was detected by CCK-8 assay. The cell growth curves were drawn and half inhibitory concentration (IC(50)) values were calculated. The cell apoptosis were detected by flow cytometry (FCM), the expression of the caspase-9, caspase-3, cleaved caspase-3, Bcl-2, Bcl-XL, Bid, Bax and c-myc protein in SUDHL-4 cells treated with IL-21 by western blot, the mRNA expression of Bcl-2, Bcl-XL, Bid, Bax, c-myc by Survivin gene with RT-PCR.

RESULTSIL-21 markedly inhibited SUDHL-4 cell growth in a time- and dose-dependent manner. The 48 hIC(50) was 140.9ng/ml; The FCM showed that the apoptosis proportion of SUDHL-4 cells treated with 100 ng/ml of IL-21 apoptosis (AnnexinV-FITC(+) positive cells) gradually increased (48 h: 19.7 ± 2.3%). The protein expression of caspase-9, caspase-3, Bcl-2 and Bcl-XL decreased in a time-dependent manner. The Bax and c-myc protein markedly increased, but the Bid protein level did not change. IL-21 up regulated c-myc and Bax gene expression, however down regulated Bcl-2 and BCL-XL gene expression, but the gene expression of Bid and Survivin hadn't been changed significantly.

CONCLUSIONSIL-21 can inhibit proliferation and induce apoptosis of SUDHL-4 cell. The mechanism may involve in endogenous mitochondrial pathway mediated by the c-myc and the Bcl-2 genes.

Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Caspase 9 ; metabolism ; Cell Line, Tumor ; Humans ; Interleukins ; administration & dosage ; pharmacology ; Lymphoma, Large B-Cell, Diffuse ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; bcl-X Protein ; metabolism

This study was aimed to investigate the impact of specific siRNA on survivin gene in transfected lymphoma cell line and provide experimental evidences for future treatment of mantle cell lymphoma. The small interfering RNA (siRNA) targeted survivin mRNA was synthesized in vitro and was transfected into Jeko-1 that showed high survivin expression in mRNA level. The levels of survivin mRNA and protein expression were detected by quantitative reverse transcription polymerase chain reaction (RT-PCR) and Western blot respectively. The apoptosis effect was examined by calculating the ratio of Annexin V-FITC/PI positive cells using flow cytometry. The inhibition of cell proliferation was assayed with CCK-8 reagent after transfection. The results showed that expression of survivin mRNA was markedly suppressed by the siRNA. The relative expression levels were 0.49 ± 0.03, 0.38 ± 0.02 and 0.17 ± 0.02 at time points of 24, 48 and 72 h respectively, compared with the control group; the inhibitive rates of cell proliferation were (31.2 ± 2.1)%, (43.3 ± 3.4)% and (52.6 ± 2.5)%; the apoptotic rates of cells were (6.3 ± 0.5)%, (13.5 ± 1.1)% and (23.6 ± 1.6)% respectively; survivin protein expression levels were gradually reduced. It is concluded that the siRNA targeting survivin down-regulates the expressions of survivin mRNA and protein evidently. The siRNA of survivin displays the potent ability to inhibit the proliferation of lymphoma cell line Jeko-1; survivin may become a potential molecular target for the therapy of lymphoma in the future.
Cell Line, Tumor ; Cell Proliferation ; Gene Silencing ; Humans ; Inhibitor of Apoptosis Proteins ; genetics ; RNA, Messenger ; genetics ; RNA, Small Interfering ; genetics ; Transfection

Objective To investigate the effect of peptide Tat-GluR6-9c on phosphorylations and protein expressions of mixed-lineage kinase 3 (MLK3),mitogen-activated protein kinase kinase 7 (MKK7) and c-Jun NH2-terminal kinase (JNK),and its effect on hippocampus CA1 region neuronal cell injury induced by cerebral ischemia followed by reperfusion.Methods Twenty four adult male SD rats were randomly divided into sham-operated group,ischemia-reperfusion group (I/R),Tat-GluR6-AA treatment group and Tat-GluR6-9c treatment group (n=6).Four-vessel occlusion method was employed to establish the cerebral ischemia models in the later 3 groups.The effects of peptide Tat-GluR6-9c on the phosphorylations and protein expressions of MLK3 (6 h after the reperfusion) and JNK (3 d after the reperfusion) were detected by Western blotting; the effects ofpeptide Tat-GluR6-9c on the phosphorylation and protein expression of MLK7 (1 d after the reperfusion) were detected by Western blotting and immunohistochemistry.Cresyl Violet (CV) staining was employed to examine the survival of CA1 pyramidal cells in the hippocampus.Results The phosphorylation of MLK3,MKK7 and JNK in Tat-GluR6-9c treatment group was significantly less than that in I/R group and Tat-GluR6-AA treatment group (P＜0.05).As compared with I/R group and Tat-GluR6-AA group,peptide Tat-GluR6-9c group could obviously increase the number of neuron cells (P＜0.05).Conclusion Peptide Tat-GluR6-9c has a protective effect on neuron in CA1 region of hippocampus following cerebral ischemia-reperfusion by significantly decreasing the phosphorylations ofMLK3,MKK7 and JNK.

To explore anti-tumor active components of Chimonanthus salicifolius, the phytochemistry of the chloroform fraction from leaves extract was investigated by repeated silica gel column chromatography. Twelve compounds were isolated and their structures were identified by physicochemical properties and spectroscopic data analysis as 9-epi-blumenol C(1), blumenol C(2), (+)-dehydrovomifoliol (3), (+)-vomifoliol (4), robinlin (5), (-)-loliolide (6), isofraxidin (7), scopoletin (8), 6,7-dimethoxycoumarin (9), 6, 7, 8-trimethoxycoumarin (10), beta-sitostenone (11), and beta-stigmasterol(12). Compounds 1-6 belonging to nor-sesquiterpenoids were isolated from the family Calycanthaceae for the first time. Compound 1 was a new natural product. Compounds 7, 11 and 12 were obtained from this plant for the first time.
Antineoplastic Agents ; analysis ; isolation & purification ; Calycanthaceae ; chemistry ; Chloroform ; chemistry ; Drugs, Chinese Herbal ; analysis ; isolation & purification ; Plant Leaves ; chemistry