Cells, Cultured ; Enzyme-Linked Immunosorbent Assay ; Female ; Fetal Blood ; Humans ; Infant, Newborn ; Lipopolysaccharides ; Lymphocytes ; metabolism ; Male ; RNA, Messenger ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Suppressor of Cytokine Signaling 1 Protein ; Suppressor of Cytokine Signaling 3 Protein ; Suppressor of Cytokine Signaling Proteins ; genetics ; metabolism ; Time Factors ; Toll-Like Receptor 2 ; genetics ; metabolism ; Toll-Like Receptor 4 ; genetics ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVETo observe the effect of Tanshinone II A on the expression of epidermal growth facter (EGF) and epidermal growth facter recepter (EGFR) in human hepatocellular carcinoma cell line SMMC-7721.

METHODSThe human hepatocellular carcinoma SMMC-7721 cells cultured in vitro was treated with different concentrations of Tanshinone II A. The proliferation of the cells was measured by MTT assay, and the apoptosis of the cells was investigated by flow cytometry and cytochemical staining with Hoechst 33342. The expression of EGF and EGFR was detected by immunocytochemistry method. The levels of EGF in medium were measured by radioimmunoassay.

RESULTTanshinone II A inhibited the growth of SMMC-7721 cells remarkably in a dose-dependent manner. The inhibitory rate reached the peak (72.5%) after 0.5 microg/ml Tanshinone II A was used for 48 h, which was significantly higher than that in the controls (P<0.05). FCM analysis showed that when SMMC-7721 cells were treated with 0.5 microg/ml Tanshinone II A, the apoptosis rates for 24 h, 48 h and 72 h were (4.06+/-0.27)%, (7.58+/-0.56)% and (5.23+/-0.13)%, respectively which were markedly higher than those in the controls (all P<0.01). SMMC-7721 cell apoptosis with cell shrinkage, nuclear chromatin concentration and fragmentation as well as the formation of apoptotic bodies were observed by cytochemical staining when treated with Tanshinone II A. The immunocytochemistry showed that the expressions of EGF and EGFR were down regulated while the concentration of Tanshinone II A was increasing. The high expression rates for EGF and EGFR were 10%, 20%, respectively, and the gray scale was 181.52+/-1.63, 179.37+/-1.59, which were markedly higher than those in the controls (all P<0.05). The levels of EGF in medium measured by radioimmunoassay were decreased significantly after Tanshinone II A treatment.

CONCLUSIONTanshinone II A can inhibit cell proliferation and induce apoptosis in hepatocellular carcinoma cell line SMMC-7721, which may be related to the down-regulation of EGF and EGFR protein expression.

Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; drug effects ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; Diterpenes, Abietane ; Down-Regulation ; drug effects ; Epidermal Growth Factor ; genetics ; metabolism ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Phenanthrenes ; pharmacology ; Receptor, Epidermal Growth Factor ; genetics ; metabolism

OBJECTIVETo elucidate whether the polymorphism of asthma immune regulator gene TIM-4 is associated with the risk of childhood allergic asthma in the southwest region of China.

METHODSTIM-4 gene promoter region RS6882076 and intron RS4704727 were studied. PCR-RFLP was used to test the genotypes of two polymorphism loci among 579 cases (average 7.2 years old) of asthma and 524 controls (average 7.6 years old) in a case-control study.

RESULTSThere were significant differences in the frequency of gene types at RS4704727 site between the asthma and the control groups (P<0.01). The results of PCR-RFLP showed that the polyporphisms of RS6882076 and RS4704727 in TIM-4 gene were present in this study population. The frequency of T allele at the RS4704727 site in the asthma group was significantly lower than that in the control group (OR=1.603; 95%CI 1.304-1.971; P<0.01). There were no significant differences in the frequencies of gene types and allele at RS6882076 site between the two groups (P>0.05).

CONCLUSIONSRS4704727 polymorphism of TIM-4 gene may be associated with childhood asthma, providing a better understanding of the pathogenesis of childhood asthma in the Southwest region of China.

Asthma ; etiology ; genetics ; Child ; Female ; Humans ; Male ; Membrane Proteins ; genetics ; Polymorphism, Single Nucleotide ; Promoter Regions, Genetic

Adult ; Female ; Gastrointestinal Hemorrhage ; metabolism ; Glutathione ; therapeutic use ; Glutathione Peroxidase ; metabolism ; Humans ; Lipid Peroxidation ; Liver ; blood supply ; Liver Cirrhosis ; complications ; metabolism ; Male ; Middle Aged ; Reperfusion Injury ; prevention & control

Objective To investigate the induction of microRNA (microRNA-106a,miR-106a)on the peritoneal metastasis of human gastric cancer cell BGC-823 by regulating matrix metalloproteinase inhibitor 2 (tissue inhibitor of metalloproteinases 2,TIMP2).Methods Human gastric cancer cell line BGC-823 was cultured to the logarithmic growth phase. The cells were divided into three groups: BGC-823, BGC-823/anti-miR-106a (antagomir)and BGC-823/negative control.Real-time PCR was used to identify the effect of antagomir.Transwell assay was used to detect the cell migratory and invasive abilities of these three groups in vitro .With small incision, the cells were injected into the abdominal cavity of nude mice to prepare a xenograft model.The animals were divided into two groups:miR-antagomir and miR-NC.The tumor growth in the nude mice was generally observed and estimated.Immunohistochemistry and Western blot methods were used to detect the expression of metastasis-associated protein TIMP2 on the several abdominal organs.Results The expression level of miR-106a was down- regulated in BGC-823/anti-miR-106a group,with the fold change of 0.05±0.01,which was significantly different from that in NC group (t=-18.001,P＜0.001).In vitro exogenously silencing of miR-106a gene,the numbers of invasive and migratory cells in BGC-823/anti-miR-106a group were both significantly lower than those in BGC-823 and BGC-823/negative control groups (P＜0.001).In vivo xenograft model showed that the down-regulation of miR-106a weakened the peritoneal metastasis ability of BGC-823 cells in nude mice abdominal cavity,which was reflected by the decrease of tumor number and tumor size.With the inhibition of miR-106a,the expression of TIMP2 in miR-antagomir group was significantly higher than that in miR-NC group (P＜0.05).Conclusion BGC-823 cell has the tumorigenicity in nude mice.Silencing of miR-106a inhibits gastric cancer cell metastasis, which suggests that it has the oncogenic function.MiR-106a may induce the strengthened peritoneal metastasis ability of BGC-823 cell through acting on TIMP2.

OBJECTIVETo investigate the clinical value of different magnetic resonance (MR) pulse sequences in diagnosis of spinal metastatic tumor.

METHODSFifteen patients with clinically suspected spinal metastatic tumor were included in this study. These patients were with documented primary tumors. Four MR pulse sequences, T1-weighted spin echo (T1WI SE), T2-weighted fast spin echo (T2WI FSE), short time inversion recovery (STIR), and gradient echo 2-D multi echo data imaging combination (GE Me-2D) were used to detect spinal metastasis.

RESULTSFifteen vertebral bodies were entire involvement, 38 vertebral bodies were section involvement, and totally 53 vertebral bodies were involved. There were 19 focal infections in pedicle of vertebral arch, 15 metastases in spinous process and transverse process. Fifty-three vertebral bodies were abnormal in T1 WI SE and GE Me-2D, 35 vertebral bodies were found abnormal in T2WI FSE, and 50 vertebral bodies were found abnormal in STIR. The verges of focal signal of involved vertebral bodies were comparatively clear in T1WI SE, comparatively clear or vague in T2WI FSE, vague in STIR, and clear in GE Me-2D.

CONCLUSIONSGE Me-2D may be the most sensitive technique to detect metastases. So three sequences (T1WI SE, T2WI FSE, GE Me-2D) can demonstrate the early changes of spinal metastasis roundly.

Cervical Vertebrae ; diagnostic imaging ; Coccyx ; diagnostic imaging ; Humans ; Lumbar Vertebrae ; diagnostic imaging ; Magnetic Resonance Imaging ; methods ; Neoplasm Metastasis ; pathology ; Radiography ; Sacrum ; diagnostic imaging ; Sensitivity and Specificity ; Spinal Neoplasms ; pathology ; secondary ; Spine ; diagnostic imaging ; Thoracic Vertebrae ; diagnostic imaging

OBJECTIVETo examine the in vitro dissolution of forsythin in Forsythia suspensa powder of different particle diameter, in order to give guidance to the grinding process.

METHODHPLC was used to determine the in vitro dissolution quantity and dissolution velocity of forsythin coarse powder, fine powder and ultramicroscopic powder.

RESULTThe dissolution curves of Forsythia suspensa coarse powder, fine powder and ultramicroscopic powder were basically inconformity to Weibull distribution. Specifically, T50 was 11.8, 10.5 and 6.8 min, respectively, and Q45 was 78.22%, 81.91% and 90.76%, respectively.

CONCLUSIONThe superfine milling process can significantly increase the dissolution quantity and dissolution velocity of forsythin.

Bridged Bicyclo Compounds, Heterocyclic ; chemistry ; Chromatography, High Pressure Liquid ; Forsythia ; chemistry ; Furans ; chemistry ; Particle Size ; Powders

Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; drug effects ; Carcinoma, Hepatocellular ; metabolism ; Cell Line, Tumor ; Cyclooxygenase 2 ; metabolism ; Humans ; Oleanolic Acid ; analogs & derivatives ; pharmacology ; Saponins ; pharmacology

OBJECTIVETo study changes of TLR2 signaling pathway expression in Kupffer cells during the process of hepatic ischemia/reperfusion in a mice model and the mechanism of TLR2 signaling pathway participating in hepatic ischemia/reperfusion injury.

METHODSBALB/c mice were divided into 3 groups: sham operation (SH), ischemia/reperfusion (I/R) and GdCl3 treatment (Gd) groups. After 4 h of reperfusion, the expression of TLR2 mRNA and membrane TLR2 protein were analyzed in ischemic lobes of the livers, and in Kupffer cells isolated from ischemic lobes. The expression of NF-kappaB in ischemic lobes was also examined. Levels of endotoxin, ALT and TNFalpha were measured at the same time point.

RESULTSThe expressions of TLR2 mRNA and protein in both ischemic hepatic lobes and Kupffer cells isolated from ischemic lobes were increased in the I/R group compared to those in the SH group, as well as the expression of NF-kappaB in ischemic lobes, which was down regulated by intravenous GdCl3 treatment. Levels of ALT and TNFalpha in the portal vein were higher in the I/R group than in the SH group, which also were decreased with treatment of GdCl3. The level of endotoxin in the three groups remained constant.

CONCLUSIONTLR2 signaling pathway in Kupffer cells is activated during the process of hepatic ischemic/reperfusion injury. The activation of TLR2 signaling pathway in Kupffer cells may play a role in this process.

Animals ; Kupffer Cells ; metabolism ; Liver ; blood supply ; Male ; Mice ; Mice, Inbred BALB C ; Reperfusion Injury ; metabolism ; Signal Transduction ; Toll-Like Receptor 2 ; biosynthesis ; genetics

BACKGROUNDRestoration of blood flow to the ischemic liver lobes may paradoxically exacerbate tissue injury, which is called hepatic ischemia/reperfusion injury (IRI). Toll-like receptor 4 (TLR4), expressed on several liver cell types, and the nuclear factor-kappa B (NF-kappaB) signaling pathway are crucial to mediating hepatic inflammatory response. Because IRI is essentially a kind of profound acute inflammatory reaction evoked by many kinds of danger signals, we investigated TLR4/NF-kappaB signaling pathway activation in a murine model of partial hepatic IRI.

METHODSWild-type mice (WT, C3H/HeN) or TLR4 mutant mice (C3H/HeJ) were subjected to 45 minutes of partial hepatic ischemia followed by 1 hour, 3 hours of reperfusion. Sham group accepted the same procedure without the obstruction of blood supply. At the end of reperfusion, the compromise of liver function and the histological change of liver sections were measured as the severity of liver injury. The level of endotoxin in the portal vein was measured by limulus assay. NF-kappaB activation was determined by electrophoretic mobility shift assay (EMSA). The levels of tumor necrosis factor-alpha (TNF-alpha) and interleukin-1beta (IL-1beta) in systemic blood after hepatic IRI were assessed by enzyme-linked immunosorbent assay (ELISA).

RESULTSThe compromise of liver function and the morphological injuries in mutant mice were relieved more markedly than those in WT mice after partial hepatic IRI. NF-kappaB activation in WT mice was stronger than that in TLR4 mutant mice, and both were stronger than those in the sham operated mice (P < 0.01). Endotoxin in each group was undetectable. The levels of TNF-alpha and IL-1beta in systemic blood were elevated in both strains, but lower in the sham operated group. These mediators were significantly decreased in TLR4 mutant mice compared with those in WT mice (P < 0.01).

CONCLUSIONSThe TLR4/NF-kappaB signaling pathway may mediate hepatic IRI triggered by endogenous danger signals. Inhibition of the TLR4/NF-kappaB pathway may be a potential therapeutic target for attenuating ischemia/reperfusion-induced tissue damage in some clinical settings.

Alanine Transaminase ; blood ; Animals ; Interleukin-1beta ; biosynthesis ; Liver ; blood supply ; Mice ; Mice, Inbred C3H ; NF-kappa B ; physiology ; Reperfusion Injury ; etiology ; Signal Transduction ; physiology ; Toll-Like Receptor 4 ; physiology ; Tumor Necrosis Factor-alpha ; biosynthesis