Aim To investigate the effect of phenyle-thanol glycosides from Cistanche tubulosa(CPhGs) on the proliferation and activation of rrPDGF-BB induced HSC and their target points for resisting hepatic fibro-sis,to elucidate the molecular mechanism in molecular level, and provide basic data for the further develop-ment of new drugs. Methods HSCs were cultivated by CPhGs with different concentrations ( 0 , 3. 91 , 7. 81 , 15. 63 , 31. 25 , 62. 50 , 125. 00 , 250. 00 , and 500 mg ·L-1 ) and IC50 of CPhGs was determined. CPhGs with different concentrations ( 25 , 50 , 75 , 100 mg · L-1 ) were selected, and after the cells were stimulated with rrPDGF-BB, cell proliferation was determined by MTT. ERK1/2 ,α-SMA, c-fos, c-jun and Collagen I mRNA and Erk1/2 ,P-Erk1/2 and CollagenⅠprotein ex-pressions were assayed by RT-PCR and Western blot. Results CPhGs of ( 50 ~100 ) mg · L-1 concentra-tions groups could effectively inhibit rrPDGF-BB-medi-ated proliferation(P<0. 05) and CPhGs of(25~100) mg·L-1 concentrations groups had no significant cyto-toxicity( P >0. 05 ) . CPhGs of ( 25 ~100 ) mg · L-1 concentrations groups could inhibit ERK1/2 ,α-SMA,c-fos, c-jun and CollagenⅠmRNA levels, and also ob-viously inhibited Erk1/2 ,P-Erk1/2 and Collagen Ⅰ pro-tein expression on HSC. Conclusions CPhGs has the protective effect against hepatic fibrosis. The mecha-nism of this process may involve the interference with PDGF/ERK1/2 signaling pathway and inhibiting the activation and proliferation of HSC.

Objective To investigate the efficacy and safety of low-dose prednisone combined with methotrexate (MTX) and hydroxychloroquine (HCQ) in the treatment of rheumatoid arthritis (RA).Methods In this 12-week study,150 patients with active rheumatoid arthritis were randomly divided into two groups:prednisone group (70 cases who were received prednisone 5 ～ 10 mg/d + MTX 10 mg/w +HCQ 0.2 g/d) and control group (80 cases who were treated by Meloxicam 7.5 mg/d + MTX 10 mg/w +Leflunomide (LEF) 20 mg/d).The primary end-points were tender and swollen joint counts,visual analogue scales (VAS),and global physician and patients assessments of disease.The secondary end-points were morning stiffness time,C-reactive protein,erythrocyte sedimentation rate,the Health Assessment Questionnaire (HAQ),DAS28 and ACR20,ACR50.Results After 12 weeks,in terms of primary endpoints,tender and swollen joint counts,VAS and global physician assessments in the prednisone group were improved significantly [(4.5 ± 2.5),(3.2 ± 3.36),(21 ± 15),(24.2 ± 16.4),(20.2 ± 10.4) vs (6.4 ±5.84),(6.6±5.5),(46±14),(37.9±19.7),(34.1±12.4),P ＜0.05orP ＜0.01].In terms of secondary end-points,the prednisone group produced higher response rates [HAQ score (0.93 ± 0.52),CRP(10.2 ± 5.8) mg/L,ESR(30 ± 14) mm/h,morning stiffness time (32.0 ± 32.3) min,DAS 28 score (3.1±0.9) vs (1.22 ±0.81),(16.3±10.1)mg/L,(33±29)mm/h,(54.7±45.4)min,(4.9±1.9),P ＜0.05 orP ＜0.01].The incidence of adverse events was similar between two groups (43％ vs 49％,P ＞ 0.05).Conclusions Low-dose prednisone combined with MTX and HCQ produced rapid and relevant improvements in RA signs and symptoms.

Objective To investigate the effect of chymase on the proliferation of rat cardiac fibroblasts (CFs) and the role of transforming growth factor-?1 (TGF-?_1).Methods Cultured CFs of neonatal SD rats were isolated by trypsinization.Cell number and DNA synthesis were evaluated by MTT assay (A_(490) value) and [~3H]-deoxythy- midine [~3H]-TdR incorporation.The mRNA expression of TGF-?_1 in CFs was determined by RT-PCR.Results Chymase increased CFs numbers and [~3H]-TdR incorporation in a dose-dependent manner.The A_(490) value of CFs stimulated by 15,30 and 60 ng/mL chymase was 0.263?0.033,0.348?0.031 and 0.387?0.026,respectively, which were all significantly higher than that of control (0.201?0.019,P

OBJECTIVE To study the anti-fibrotic effect of Cistanche phenylethanoid glycosides (CPhG) in bovine serum albumin (BSA)-induced liver fibrosis in rats and its possible mechanism METHODS Seventy-five SD rats were randomly divided into six groups:normal control(distilled water-treated),model(BSA-treated),positive drug〔BSA-treated+compound Biejiarangan tablets(BJRG) 0.6 g·kg-1〕,and BSA-treated+CPhG(0.125,0.25 and 0.5 g·kg-1)groups. There were thirteen rats in each BSA-treated+CPhG(0.125,0.25 and 0.5 g·kg-1)group and twelve rats in other groups. Subcutaneous injection and tail vein injection of BSA immunity were used to induce the rat liver fibrosis model. Meanwhile, different therapeutic drugs were ig adminstered to rats. After the experimental period,rats were fasted for 12 h prior to 10%chloral hydrate administration and immediately euthanized. The liver was weighed to calculate the liver index. Glutamic-pyruvic transaminase (GPT),glutamic-oxalactic transaminase (GOT),alkaline phosphatase(ALP),total protein(TP)and albumin(ALB)were evaluated by the Mind-Ray automatic biochemical analyzer. The density of hydroxyproline (HyP) in liver tissues was determined using a spectrophotometric method according to the kit′s instructions. Histopathological changes and expressions of typeⅠ and typeⅢcollagens in liver tissues were also determined by immunohisto?chemical staining. RESULTS Compared with the normal control group,collagen fibers of liver tissues in the model group extended their links and enveloped the entire lobule,causing lobular structural damage and the formation of pseudolobules. The liver index(P<0.05),GPT,GOT,ALP,TP and ALB serum levels(P<0.05),HyP content(P<0.01)were significantly increased,so was the expression of typeⅠcollagens and typeⅢcollagens(P<0.01)in the model group. Compared with model group,various doses (0.125,0.25 and 0.5 g · kg-1) of CPhG significantly reduced the BSA-induced elevation of the liver index;GPT,GOT,ALP,TP and ALB serum levels(P<0.05),and HyP content decreased(P<0.01);the morphology of the pathological tissue sections was close to that of the normal control group,and CPhG significantly reduced the expression of two types of collagens(P<0.01). CONCLUSION CPhG can significantly reduce the degree of BSA-induced liver fibrosis in rats. The mechanism may be associated with down-regulation of two types of collagens and suppression of the activation of hepatic stellate cells.

The fine structure of enoxaparin sodium samples with different degree of 1,6-anhydro derivatives were analyzed with polyacrylamide gel electrophoresis, high performance liquid chromatography, ultraviolet spectroscopy, infrared spectroscopy and nuclear magnetic resonance spectroscopy. A further study of anticoagulation activity of enoxaparins was performed, including those on their inhibition activities of coagulation factor Xa (FXa) and thrombin (FIIa). The results showed that the anti-FXa and -FIIa activities of enoxaparins with different degree of 1,6-anhydro derivatives (20.0%-39.7%) with similar structure characteristics, had decreasing tendency when the degree of 1,6-anhydro derivatives increased. Especially, the anti-FXa activity was sensitive to the change of the degree of 1,6-anhydro derivatives.
Anticoagulants ; chemistry ; Enoxaparin ; chemistry ; Factor Xa Inhibitors ; chemistry ; Thrombin ; antagonists & inhibitors

0.05)between the two groups.Conclu- sion The long-term outcomes of e-CHB is not markedly different compared with that of e+CHB.

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

OBJECTIVEAs our previous comparative proteomics study on high and low metastasis human hepatocellular carcinoma (HCC) cell strains revealed that cytokeratin 19 (CK19) was related to higher metastasis potential, we further investigated the relationship between serum CK19 level in HCC patients and their clinico-pathologic characteristics.

METHODSSerum CK19 levels of 101 normal controls and 108 pathology-proven HCC patients were determined using radioimmunoassay, and the their correlation with clinico-pathologic parameters were studied.

RESULTSThe upper limit of one-side 98% confidence interval of normal serum CK19 level was 2.3 microg/L. Among 108 HCC patients, 24 (22.2%) had increased serum CK19 level, ranging from 2.4 to 45.5 microg/L. There were 12 patients (11.1%) with increased CK19 level but normal AFP level. The percentage of poor differentiated tumor was higher in CK19 increased cases (37.5%, 9/24) than in CK19 normal cases (20.2%, 17/84). Moreover, the presence of portal vein tumor emboli was significantly higher in CK19 increased cases (25.0%, 6/24) than in CK19 normal cases (6.0%, 5/84). (Chi-square = 7.403, P < 0.01) In addition, the percentage of TNM stage III/IV tumor was significantly higher in CK19 increased patients (54.2%, 13/24) than in CK19 normal cases. (chi-square = 13.300, P < 0.005)

CONCLUSIONSome HCC patients do have increased serum CK19 level, which could be related to portal vein tumor emboli, poor tumor differentiation and advanced tumor stages.

Adult ; Aged ; Biomarkers, Tumor ; blood ; Carcinoma, Hepatocellular ; blood ; pathology ; Female ; Humans ; Keratins ; blood ; Liver Neoplasms ; blood ; pathology ; Male ; Middle Aged ; Neoplasm Proteins ; blood ; Peptide Fragments ; blood ; genetics ; Proteome ; analysis

Bronchial Diseases ; diagnosis ; Child ; Female ; Humans ; Infant ; Male ; Tuberculosis, Pulmonary ; diagnosis

OBJECTIVESTo examine the activities of transcription factors (TFs) in human hepatocellular carcinoma (HCC) cell lines with different metastatic potentials, so as to identify the TFs associated with HCC metastasis.

METHODSTranscription factor activity profile of Hep3B, MHCC97L and MHCC97H, three HCC cell lines with different metastatic potentials, were examined using protein/DNA array. Electrophoretic mobility shift assays (EMSA) and Western blot were used to confirm the results obtained by protein/DNA array.

RESULTSFrom a total of 345 screened TFs, 7 activity differential TFs were found, of which 5 showed increased activity, including p53, hypoxia inducible factor-1 alpha (HIF-1alpha), signal transducer and activator of transcription 3 (Stat3) and Sp1, and 2 showed decreased activity including Rb and Smad3.

CONCLUSIONThe abnormal functioning of transcription factors is closely associated with HCC metastasis. Our present findings could be of help in expanding our understanding of the mechanism of HCC metastasis and identify new predictive biomarkers and therapeutic targets.

Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Line, Tumor ; DNA Fingerprinting ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Neoplasm Metastasis ; Protein Array Analysis ; Transcription Factors ; classification ; genetics ; metabolism