To investigate the effects of gambognic acid (GA) on TRAIL-induced apoptosis of cancer cells, human colon HT-29 cancer cells were treated with GA to promote apoptosis. Inhibition of the cell proliferation was measured with MTT assay and cell apoptosis was detected with formation of DNA ladders in agarose gel electrophoresis, and activation of caspase activity. The content of cytosolic reactive oxygen species (ROS) was measured with flow cytometry. The activities of Caspase-3, -8, -9 were detected using spectrophotometric assay. The levels of c-FLIP, CHOP, DR4 and DR5 in cells were tested by Western blot. Combination of GA (1 µg · mL(-1)) and TRAIL (40 ng · mL(-1)) significantly reduced proliferation and increased apoptosis of HT-29 cells over those induced by each agent alone. Percentage of apoptotic cells was increased to 45.5%. GA markedly enhanced the intracellular ROS generation. Expression of CHOP, DR4 and DR5 was up-regulated to 7.38, 5.41, and 4.85 times of the control group, respectively. GA promoted activation of Caspase-3, -8, and -9 by TRAIL (P<0.05). Furthermore, the expression of anti-apoptotic protein c-FLIP was down-regulated to 0.22 ± 0.08 times of the control group. In conclusion, GA sensitizes HT-29 cells to TRAIL-induced apoptosis by promoting ROS-activated ERS pathways, up-regulating of DR4 and DR5, and inhibiting c-FLIP expression.
Apoptosis ; Apoptosis Regulatory Proteins ; metabolism ; Caspases ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; Colonic Neoplasms ; metabolism ; Down-Regulation ; HT29 Cells ; Humans ; Reactive Oxygen Species ; metabolism ; TNF-Related Apoptosis-Inducing Ligand ; pharmacology ; Up-Regulation ; Xanthones ; pharmacology

OBJECTIVETo investigate the clinical features of Crohn disease according to the Montreal classification.

METHODSClinical data of 43 surgical patients with Crohn disease (surgical group) and 125 non-surgical patients with Crohn disease (non-surgical group) were retrospectively analyzed and compared between two groups. The Montreal classification was used.

RESULTSIn the surgical group, 28 patients (65.1%) were A2, 14 (32.6%) were A3 and only one was A1, which was not significantly different as compared to the non-surgery group. The proportions of L1, L2, L3, and L4 subtype in the surgical group were 41.9%, 25.6%, 30.2%, and 2.3%, respectively, which was not significantly different as compared to that in the non-surgery group. In the surgical group,B1 disease was found in 1 case (2.3%), B2 in 26 cases (60.5%), and B3 in 16 cases (37.2%), while in the non-surgical group, B1 was found in 79 cases (63.2%), B2 in 44 cases (35.2%) and B3 in 2 cases (1.6%). Differences were significant between two groups in disease behavior (P=0.001, P=0.004, P=0.001).

CONCLUSIONSMost surgical patients of Crohn disease are A2. L1 and L3 are the main lesion location. As disease behavior, B2 and B3 are the main reasons for operation.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Crohn Disease ; classification ; pathology ; surgery ; Female ; Humans ; Male ; Middle Aged ; Reference Standards ; Retrospective Studies ; Young Adult

OBJECTIVETo search the forming cause and the correlation between the clinical phenotype and chromosome band by the cytogenetic analysis on a case of ring chromosome 21 syndrome.

METHODSIdentification and location of 21 ring chromosome were performed with the G-banding, C-banding, N-banding, high-resolution banding and fluorescence in situ hybridization (FISH) techniques.

RESULTSIt was found that the karyotypes of the patient's parents are normal. The patient's karyotype is 46,XY, r(21)[91]/46,XY,r(21;21)(p11q22.3;p11q22.3) [5]/45,XY,-21[4].

CONCLUSIONThe clinical phenotype of ring chromosome 21 syndrome is related to the deletion of distal segment of 21q, and the abnormal sexual development of male is related with the deletion of 21q22.3.

Child, Preschool ; Chromosome Aberrations ; Chromosome Disorders ; genetics ; pathology ; Chromosomes, Human, Pair 21 ; genetics ; Cytogenetic Analysis ; methods ; Humans ; In Situ Hybridization, Fluorescence ; Karyotyping ; Male ; Phenotype ; Ring Chromosomes ; Syndrome

OBJECTIVETo compare different expression profiles of all known chemokine receptors in human hepatocellular carcinoma (HCC) cell lines with different metastasis potentials.

METHODSEighteen pairs of chemokine receptor primers were designed using Premier software. Expression profiles of the 18 chemokine receptors on four HCC cell lines of lower to higher potentials of metastasis (SMMC-7721, MHCC97-L, MHCC97-H and HCCLM6) were analyzed by RT-PCR. Expression of CXCR4 was detected by RT-PCR.

RESULTSExpression profiles of chemokine receptors on four HCC cell lines with different metastatic potentials had significant differences (P < 0.01), in which CCR10, CXCR4 and CXCR6 expressions decreased gradually as the metastatic potential of the cell lines increased. The expressions of CCR3, CCR4, CCR10, CCR12 and XCR1 on HCCLM6 were significantly reduced compared with SMMC-7721 (P < 0.01), whereas the expressions of CXCR1 (P = 0.006) and CXCR5 (P = 0.003) exceeded that of SMMC-7721. Except for CXCR2, CXCR6 and XCR1, most of chemokine receptors on MHCC97-H were expressed differently compared with MHCC97-L (P < 0.05), in which expressions of CCR1 (P = 0.002), CCR2 (P = 0.004) and CCR5 (P = 0.046) exceeded MHCC97-L. CXCR4 was detected only on the positive controls and SMMC-7721 when the template of total RNA was reduced one-half in RT-PCR.

CONCLUSIONChemokine receptors are expressed very differently at mRNA level on HCC cell lines with different metastatic potentials. The different profiles of chemokine receptors in tumor microenvironment and the function of CXCR4 in HCC should be further studied. Our findings have important implications in understanding the relationship between chemokine receptors and the metastatic potential of HCC.

Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Line, Tumor ; Humans ; Liver Neoplasms ; metabolism ; pathology ; RNA, Messenger ; genetics ; Receptors, Chemokine ; metabolism

Objective To observe the clinical efficacy and safety of deprogesterone tablets in the treatment of climacteric syndrome.Methods A total of 86 women with climacteric syndrome were randomly divided into control group and treatment group with 43 cases per group.Control group received placebo 10 mg per time,qd,orally.Treatment group was treated with deprogesterone 10 per time,qd,orally.Two groups were treated for one month.The clinical efficacy,the levels of sex hormone and blood lipid,adverse drug reactions were compared between two groups.Results After treatment,the total effective rates of treatment and control groups were 93.02％ (40 cases/43 cases) and 72.09％ (31 cases/43 cases) with statistically significant difference (P ＜ 0.05).After treatment,the main indexes in treatment and control groups were compared:luteinizing hormone were (11.70 ± 1.13) and (28.36 ± 4.48) U · L-1,follicle stimulating hormone were (36.31 ±4.24) and (60.77 ± 7.75) U · L-1,estradiol were (71.76±9.37) and (45.43 ±5.21)pmol · L-1,triglyceride were (1.23 ± 0.24) and (1.72 ± 0.20) mmol · L-1,total cholesterol were (4.94 ± 0.66) and (5.82 ± 0.58) mmol · L-1,low density lipoprotein cholesterol were (2.64 ± 0.36) and (3.25 ±0.31)mmol · L-1,high density lipoprotein cholesterol were (1.66 ±0.21) and (1.38 ±0.13) mmol · L-1,the differences were statistically significant (all P ＜ 0.05).The adverse drug reactions in treatment group were based on insomnia (2 cases) and headache (2 cases).And there was no adverse drug reaction in the control group.The incidences of adverse drug reactions in treatment and control groups were 9.30％ and 0 without significant difference (P ＞ 0.05).Conclusion Deprogesterone tablets have a definitive clinical efficacy in the treatment of climacteric syndrome,which can effectively adjust the levels of sex hormone and blood lipid,without increasing the incidence of adverse drug reactions.

OBJECTIVETo study the clinical characteristics of hospitalized infants with allergic proctocolitis, and to provide a scientific basis for early diagnosis and effective treatment of allergic proctocolitis.

METHODSThe clinical data of 96 infants with allergic proctocolitis between September 2011 and March 2013 were reviewed retrospectively. Descriptive analysis was performed to assess the clinical characteristics of allergic proctocolitis.

RESULTSThe most common clinical manifestation was diarrhea in the 96 infants. The electronic colonoscopy results indicated that 40% of the infants had multiple small nodules, 26% showed focal erythema and brittle mucous membranes, 25% showed multiple superficial erosion, and 9% showed ulcers with surface exudates. The affected areas included the sigmoid colon (87%), rectum (24%), descending colon (13%), and transverse colon ascending colon and ileocecal junction (8%). Histopathologic examination showed eosinophilic infiltration of mucosal layers, the condition of which was mild to moderate in 89% and severe and extremely severe in 12% of the infants. To treat the allergic proctocolitis, mothers and infants were suggested to avoid allergenic foods; 43% of them continued breastfeeding, 45% switched to highly hydrolyzed protein formula, and 13% were prescribed amino acid-based elemental formula. All infants were in complete remission at discharge.

CONCLUSIONSAs the clinical manifestations of allergic proctocolitis in infants lack specificity, the electronic colonoscopy and mucosal histopathologic examination are helpful for early and differential diagnosis. The best treatment is to avoid allergenic foods. Formula-feeding infants should be prescribed highly hydrolyzed protein formula or amino acid-based elemental formula.

Colonoscopy ; Female ; Humans ; Infant ; Male ; Proctocolitis ; diagnosis ; pathology ; therapy ; Retrospective Studies

OBJECTIVESTo study the relationship between lymphangiogenesis and lymphatic metastasis in mice bearing hepatic carcinoma and analyze the mechanism of the lymphatic metastasis.

METHODSHepatic carcinoma cell lines of high and low potentialities of lymphatic metastasis were injected into the footpads of Balb/c mice. Their metastases to lymph nodes were examined. The tumor tissues of each group were stained with 5'-nucleotidase-ALP to observe the lymphoangiogenesis. The total RNA of high and low metastatic potential cell lines were extracted for metastasis gene DNA array. The vascular endothelial cell growth factor C (VEGF-C) and VEGF-D of each cell line were detected using semi-quantitative RT-PCR and were further quantatively analyzed using real time PCR.

RESULTSThe para-common iliac a. and renal hilar lymph nodes metastases of the high metastatic potential cells were significantly higher than in the controls (P>0.05). The quantity of lymphatic vessels in the high metastasis group was significantly larger than that of the control group (P<0.05). The expressions of CD44, E-cadherin, HER2/neu, H-Ras and VEGF-C in the high metastasis group were higher than those in the low metastasis group shown by the cDNA micro array experiment but the expressions of nm23A, nm23-E4, p16ink4a, CD61 were lower. The VEGF-C expression was higher and the VEGF-D was lower in the high metastasis group compared to those of the low metastasis group shown by semi-quantitative RT-PCR. The secretion of VEGF-D was significantly lower and the ratio of VEGF-C/VEGF-D was significantly higher in the high metastasis group than the low metastasis group (P<0.05).

CONCLUSIONSThe lymphatic metastasis of hepatic carcinoma is related to lymphoangiogenesis. The changes of VEGF-C and VEGF-D expressions might be a cause influencing the lymphoangiogenesis. VEGF-C/VEGF-D might be an effective parameter in affecting lymphatic metastases.

Animals ; Liver Neoplasms, Experimental ; pathology ; Lymph Nodes ; pathology ; Lymphangiogenesis ; Lymphatic Metastasis ; Male ; Mice ; Mice, Inbred BALB C ; Neoplasm Transplantation ; Vascular Endothelial Growth Factor C ; metabolism ; Vascular Endothelial Growth Factor D ; metabolism

OBJECTIVETo study the effects of osteopontin (OPN) on the phenotypes of human hepatocellular carcinoma cell line SMMC-7721.

METHODSHuman hepatocellular carcinoma SMMC-7721 cells were transfected with plasmid pcDNA 3.1(-)/OPN and cells transfected with a mock plasmid served as controls. OPN expression was verified by RT-PCR and Western blot, and concentrations of OPN, MMP-2, MMP-9 and uPA were measured by ELISA. A series of functional assays in vitro were used to monitor the changes of SMMC-7721 malignant phenotypes.

RESULTSOPN expression of SMMC-7721 cells was elevated after transfection. Concentrations of OPN, MMP-2 and uPA in the medium of SMMC-7721 cells after transfection were higher than those of the controls [(3.02+/-0.12) ng/ml vs (1.43+/-0.07) ng/ml, (43.04+/-3.06) ng/ml vs (22.15+/-4.34) ng/ml, and (4.78+/-0.70) ng/ml vs (1.61+/-0.34) ng/ml respectively, P less than 0.01], but MMP-9 concentration did not increase [(7.82+/-2.25) ng/ml vs (7.70+/-1.92) ng/ml]. Functional assays in vitro indicated that SMMC-7721 cells after transfection showed higher adhesive, migrant and invasive capabilities than those of the controls (cell adhesion rates were 75.33%+/-10.59% vs 57.34%+/-2.52%; number of outer surface cells in migrant assay was 14.33+/-2.51 vs 6.34+/-1.53; cell number in the invasive assay was 8.23+/-1.53 vs 4.12+/-1.29 respectively, P less than 0.05).

CONCLUSIONOPN might enhance the expression of MMP-2 and uPA to promote malignant phenotypes of SMMC-7721 cells.

Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Line, Tumor ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Matrix Metalloproteinase 2 ; secretion ; Osteopontin ; genetics ; metabolism ; Transfection ; Urokinase-Type Plasminogen Activator ; secretion

OBJECTIVETo observe the effects of Tianma Gouteng Fang (TGF) on the transmitter amino acids in the hippocampus extracellular liquids in freely moving rats subjected to incomplete brain ischemia.

METHODHippocampus extracellular liquids was collected continuously by the microdialysis sampling technology in freely moving rats during pre-ischemia, incomplete ischemia and reperfusion periods induced by the occlusion and loose of both common carotid arteries. Each dialysate sample was assayed for GABA, Tau, Glu, Cys and Arg with HPLC-electrochemical detector.

RESULTTGF increased the concentrations of GABA and Tau in the extracellular liquids of rat hippocampus. Compared with the model group, the concentration of Glu in the middle and large dosage groups of TGF, during the 120 min of ischemia, reduced by 38.64% and 31.35%, Tau increased by 13.99% and 12.86%, GABA advanced 25.89% and 33.99%, Cys decreased by 40.93% and 42.08%, Arg raised to 116.95% and 108.96%, respectively. After 120 min of reperfusion, the concentration of Glu decreased by 14.55% and 11.48%, Tau increased by 16.13% and 14.03%, GABA increased by 24.41% and 26.22%, respectively.

CONCLUSIONTGF can increase the concentration of inhibitory amino acids in hippocampus extracellular liquids of rats and inhibit the excessive release of excitatory amino acids and raise the concentration of the inhibitory amino acids and Arg during the ischemia-reperfusion periods. Therefore, TGF can play the neuroprotective role.

Animals ; Arginine ; metabolism ; Brain Ischemia ; complications ; metabolism ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Excitatory Amino Acids ; metabolism ; Extracellular Space ; metabolism ; Gastrodia ; chemistry ; Hippocampus ; metabolism ; Male ; Neuroprotective Agents ; pharmacology ; Plants, Medicinal ; chemistry ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; etiology ; metabolism ; Taurine ; metabolism ; Uncaria ; chemistry ; gamma-Aminobutyric Acid ; metabolism

OBJECTIVEWe previously showed that introduction of a normal, neomycin-tagged human chromosome 8 reduced the metastatic capacity of C5F rat liver cancer cell line, which had high metastatic potential without affecting tumorigenicity, suggesting the presence of one or more metastasis suppressor genes encoded on human chromosome 8. We proceeded to define further the region harboring the metastasis suppressor gene(s) and to determine the random loss of human chromosome 8 by PCR amplification of sequence tag site (STS) markers.

METHODSThe national Center for Biotechnology Information (NCBI) databases were used as references of the relative genetic distances of the STS markers. C5F genomic DNA and A9/neo8 genomic DNA were used as negative and positive controls for chromosome 8 amplification, respectively. Genomic DNA was isolated and quantified from cultured hybrid clones (A9/C5F-1 and A9/C5F-2 microcell hybrid clones served as metastasis-unsuppressed groups; A9/C5F-4, A9/C5F-8 and A9/C5F-10 microcell hybrid clones served as metastasis suppressed groups). STS-PCR products were separated by electrophoresis through 2% agarose gel.

RESULTSMetastasis-suppressed microcell hybrid clones (A9/C5F-4, A9/C5F-8 and A9/C5F-10) conserved STS markers between D8S542 --> D8S1973 (8p21.1-23.1). In contrast, metastasis-unsuppressed clones (A9/C5F-1 and A9/C5F-2) lacked several markers in this region. In attempts to refine the region retained in the microcell suppressed clones, more densely spaced STS markers in the human chromosome 8p21.1-23.1 were used. We found that the metastasis-suppressed clones retained 18cM region between D8S542 and D8S1973 (8P21.1-23.1), where as the metastasis-unsuppressed clones lacked the region.

CONCLUSIONOur results suggest that a metastasis suppressor gene is located within the interval between D8S542 and D8S1973 on human chromosome 8p21.1-23.1.

Carcinoma, Hepatocellular ; genetics ; Cell Line ; Cell Line, Tumor ; Chromosome Mapping ; Chromosomes, Human, Pair 8 ; genetics ; Fibroblasts ; cytology ; Genes, Tumor Suppressor ; Humans ; In Situ Hybridization, Fluorescence ; Liver Neoplasms ; genetics ; Neoplasm Metastasis ; Sequence Tagged Sites