Objective: To explore the protective effect of formula of Gougancai decoction (FGD) on acute liver injury induced by carbon tetrachloride (CCl₄) in rats, in order to provide basis for the development of pharmaceutical preparations or healthcare products. Method: Sixty rats were randomly divided into normal group, Silymarin group (120 mg·kg^-1) and FGD groups (475, 950, 1 900 mg·kg^-1). The normal group and the model group were given equal volume of saline by gavage, while the other groups were administered with the corresponding dose of drugs according to the body weight. After 10 days, the acute liver injury model was established with 12% carbon tetrachloride peanut oil solution (5 mL·kg^-1), except the normal group. All of the rats were put to death to collect serum and liver tissues. The contents of aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), total bilirubin (TBIL), malondialdehyde (MDA), superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) were detected by biochemical methods, the levels of tumor necrosis factor-alpha (TNF-α), interleukin-1β (IL-1β) and interleukin-6 (IL-6) in liver tissues were determined by enzyme-linked immunosorbnent assay(ELISA). Nuclear factor-κB (NF-κB) and peroxisome proliferator-activated receptor-γ (PPAR-γ) protein expression in liver tissues were detected by Western blot, and htoxylin eosin (HE) staining was used to observe the variation of liver histopathological. Result: Compared with the normal group, the serum activities of AST, ALT, ALP and the content of TBIL, MDA in the model group were significantly increased (P<0.01), the levels of TNF-α, IL-1β, IL-6 in liver tissue were remarkably increased (P<0.01), but the serum activities of SOD, GSH-Px were significantly decreased (P<0.01), the expression of NF-κB was enhanced in liver tissue (P<0.01), and PPAR-γ was down-regulated (P<0.01), indicating the successful modeling of acute liver injury. Compared with the model group, FGD could reduce the activities of AST, ALT, ALP and the contents of TBIL, MDA (P<0.05, P<0.01), decease the level of TNF-α, IL-1β, IL-6 (P<0.05, P<0.01), and down-regulate the expression of NF-κB (P<0.05, P<0.01), but up-regulate the activities of SOD, GSH-Px and the expression of PPAR-γ (P<0.05, P<0.01). The liver tissue lesions were alleviated to varying degrees. Conclusion: FGD has a protective effect on CCl₄-induced acute liver injury in rats, and its mechanism may be related to the activation of PPAR-γ and the inhibition of NF-κB signaling pathway, with anti-inflammatory and anti-oxidative effects.

OBJECTIVE: To explore the transcriptional regulation mechanism and biological function of low expression of vasoactive intestinal peptide receptor 1 (VIPR1) in hepatocellular carcinoma (HCC). METHODS: We constructed plasmids carrying wild-type VIPR1 promoter or two mutant VIPR1 promoter sequences for transfection of the HCC cell lines Hep3B and Huh7, and examined the effect of AP-2α expression on VIPR1 promoter activity using dual-luciferase reporter assay. Pyrosequencing was performed to detect the changes in VIPR1 promoter methylation level in HCC cells treated with a DNA methyltransferase inhibitor (DAC). Chromatin immunoprecipitation was used to evaluate the binding ability of AP-2α to VIPR1 promoter. Western blotting was used to assess the effect of AP-2α knockdown on VIPR1 expression and examine the differential expression of VIPR1 in the two cell lines. The effects of VIPR1 overexpression and knockdown on the proliferation, cell cycle and apoptosis of HCC cells were analyzed using CCK8 assay and flow cytometry. We also observed the growth of HCC xenograft with lentivirus-mediated over-expression of VIPR1 in nude mice. RESULTS: Compared with the wild-type VIPR1 promoter group, co-transfection with the vector carrying two promoter mutations and the AP-2α-over-expressing plasmid obviously restored the luciferase activity in HCC cells (P < 0.05). DAC treatment of the cells significantly decreased the methylation level of VIPR1 promoter and inhibited the binding of AP-2α to VIPR1 promoter (P < 0.01). The HCC cells with AP-2α knockdown showed increased VIPR1 expression, which was lower in Huh7 cells than in Hep3B cells. VIPR1 overexpression in HCC cells caused significant cell cycle arrest in G2/M phase (P < 0.01), promoted cell apoptosis (P < 0.001), and inhibited cell proliferation (P < 0.001), while VIPR1 knockdown produced the opposite effects. In the tumor-bearing nude mice, VIPR1 overexpression in the HCC cells significantly suppressed the increase of tumor volume (P < 0.001) and weight (P < 0.05). CONCLUSION VIPR1 promoter methylation in HCC promotes the binding of AP-2α and inhibits VIPR1 expression, while VIPR1 overexpression causes cell cycle arrest, promotes cell apoptosis, and inhibits cell proliferation and tumor growth.
Animals ; Carcinoma, Hepatocellular/pathology* ; Cell Line, Tumor ; Cell Proliferation/genetics* ; Gene Expression Regulation, Neoplastic ; Humans ; Liver Neoplasms/pathology* ; Luciferases/genetics* ; Methylation ; Mice ; Mice, Nude ; Receptors, Vasoactive Intestinal Polypeptide, Type I/metabolism* ; Transcription Factor AP-2/metabolism*

OBJECTIVETo investigate anticancer effects of 5-fluorouracil (5-FU) combined with CL, extract of Rosa roxburghii Tratt on human endometrial adenocarcinoma cell line (JEC).

METHODSJEC cells cultured in vitro in the logarithmic growth phase were seeded in the culture plate and divided into the control group (RPMI 1640), the positive group (10(-4) mol/L 5-FU), the CL groups (at the dose of 0.01, 0.1, 1, 10, and 100 microg/mL), and the CL (0.01, 0.1, 1, 10, and 100 microg/mL) combined with 5-FU groups. Effects of 5-FU combined with CL on JEC cell growth were drawn and measured by MTT and growth curves. Effects of CL combined with 5-FU on the JEC cell differentiation was analyzed by detecting the reduction capability of nitrobenzene thiocyanate (NBT) and lactate dehydrogenase (LDH) contents in the cultured medium. Effects of CL combined with 5-FU on the JEC cell apoptosis and cell proliferation cycle were detected by acridine orange (AO)/ethidium bromide (EB) fluorescent staining and flow cytometry (FCM).

RESULTSThe proliferation inhibitory effect of CL combined with 5-FU on JEC cells was enhanced when compared with that of CL or 5-FU alone (P<0.05). The percentages of NBT positive JEC cells and apoptotic JEC cells increased in the 5-FU combined with CL groups when compared with 5-FU group or the CL group alone (P<0.05). The LDH concentration of the JEC cell culture supernate decreased in 5-FU combined with CL groups (P<0.05). Furthermore, the percentage of G0-G1 phase JEC cells treated by 5-FU combined with CL was higher than that of 5-FU or CL alone (P<0.05).

CONCLUSIONCL could enhance anticancer effects of 5-FU. Its mechanisms might be correlated with reinforcing the cytotoxicity of 5-FU, inducing cell differentiation and apoptosis, and inhibiting cell proliferation and division.

Adenocarcinoma ; pathology ; Cell Differentiation ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Endometrial Neoplasms ; pathology ; Female ; Fluorouracil ; pharmacology ; Herb-Drug Interactions ; Humans ; Plant Extracts ; pharmacology ; Rosa ; chemistry

In the past, liver cirrhosis was thought to be irreversible. However, theoretically, the process of liver cirrhosis could be slowed down by inhibiting extracellular matrix (ECM) production and by promoting its degradation. Wnt signaling pathway has been involved in many physiological and pathological processes and some studies have confirmed that abnormal activation of Wnt signaling pathway could cause activation of hepatic stellate cells, and then further induce ECM formation and prompt cirrhosis progression. We believe that new clinical therapeutic direction would emerge after in-depth exploration of the mechanism of Wnt signaling pathway involved in liver cirrhosis progression.

Liver fibrosis has a complex pathogenesis and involves various intracellular and extracellular signal transduction pathways, but its specific pathogenesis remains unclear. Recent studies have confirmed that the activation of hepatic stellate cells due to abnormal activation of the Wnt signaling pathway is an important cause of liver fibrosis, and targeted inhibition of the Wnt signaling pathway can prevent the activation and proliferation of hepatic stellate cells and thus achieve an antifibrogenic effect. This article reviews the role of the Wnt signaling pathway in the development and progression of liver fibrosis and the latest research advances in the antifibrogenic effect of the interference of the Wnt signaling pathway, in order to provide new ideas for anti-hepatic fibrosis and the delay of early-stage cirrhosis.

OBJECTIVETo observe the therapeutic effect of parthenolide (PTL) on rabbit knee arthritis (KOA) and its effects on serum expression of interleukin-1beta (IL-1beta) and contents of tumor necrosis factor-alpha (TNF-alpha).

METHODSEight rabbits were randomly selected from 40 healthy pure-bred New Zealand rabbits as the normal control group. The KOA model was established in the rest 32 rabbits by plaster cast fixation of the right hind limb extension position. After modeling they were randomly divided into 4 groups, i.e., the model control group, the high dose PTL group, the middle dose PTL group, and the low dose PTL group, 8 in each group. Serum contents of IL-1beta and TNF-alpha were detected using enzyme-linked immunosorbent assay.

RESULTSCompared with the model group, IL-1beta and TNF-alpha concentration decreased in the 3 PTL groups (P < 0.01). The decrement was positively correlated with PTL concentrations (IL-1beta: r = 0.55, P < 0.01; TNF-alpha: r = 0.56, P < 0.01). The inhibition reached the peak when the PTL concentration arrived at 20 micromol/L.

CONCLUSIONSPTL could down-regulate the blood IL-1beta and TNF-alpha concentrations of KOA rabbits. Besides, the decrement was positively correlated with the PTL concentration.

Animals ; Female ; Interleukin-1beta ; blood ; Male ; Osteoarthritis, Knee ; blood ; drug therapy ; Phytotherapy ; Rabbits ; Sesquiterpenes ; pharmacology ; therapeutic use ; Tumor Necrosis Factor-alpha ; blood

This work was launched to study on the chemical constituents from Euphorbia thymifolia. Thirteen compounds were isolated from the 95% ethanol extract of the aerial parts of E. thymifolia by column chromatographies on silica gel, Sephadex LH-20, MCI, and ODS, and preparative HPLC, including two thymol derivatives(1-2), four alkaloids(3-6), five isocoumarins(7-11), together with two ellagic acids(12-13). All the compounds are listed as follows:(Z)-8,9-dehydro-9,10-diisobutyryloxythymol(1), 8-hydro-xy-9,10-diisobutyryloxythymol(2), N-(N-benzoyl-L-phenylalanyl)-L-phenylalanol(3), aurantiamide acetate(4), 1-carboethoxy-β-carboline(5), isoechinulin A(6), ethyl brevifolincarboxylate(7), euphorhirtin B(8), 4,5-didehydro chebulic acid triethyl ester(9), euphorhirtin G(10), pomegranatate(11), 3,3',4'-tri-O-methylellagic acid(12), 3,3'-di-O-methylellagic acid(13). Compound 1 is a new compound. Except for compound 4, the others were isolated from this plant for the first time. All the compounds were screened for anti-neuroinflammatory activity in vitro, and compounds 1-3 and 7 showed significant activity with IC_(50) values of 0.19,12.93,7.29,25.4 μmol·L~(-1).
Chromatography, High Pressure Liquid ; Euphorbia

From the standpoint of training mode,the paper makes an effort to analyze the present situation on Sino-Foreign cooperation in nnning schools in mode of nursing personnel training in Hubei Province,find out the existing problems and put forward the corresponding proposals in order to ensure sustainable development of the Sino-Foreign cooperation in running schools in mode of nursing personnel training.

Objective:To construct a phage display antibody library of special antigen responding to serum starved U251 cell,from which the serum responding gene and protein would be gotten.Methods:U251 cells were cultured in serum-absent midium for 48 h.Its protein was extracted and used to immunize BALB/c mice.Total RNA of the spleenocytes of immunized mice was extracted.VH and VL were amplified by RT-PCR and were linked into ScFv(Single chain fragment of variation) with a linker.ScFv was recombined to pCANTAB5E vector and was transformed to TG1 strain.Results:The library capacity was up to 3?106 cfu/L.A positive clone was identified from 8 random clones of this library.Conclusion:The special ScFv phage library is constructed successfully.It is the basis for screening special antibodies which can recognize serum responding protein.

OBJECTIVETo study on CTA and MRI in the diagnosis and treatment of hemangioma in the limb skeletal muscles, and to explore therapeutic effects of surgical treatment.

METHODSFrom April 2003 to February 2011, 18 patients with intramuscular hemangioma in extremities were treated with surgical excision or dense circle suture method. Among the patients, 8 patients were male and 10 patients were female,ranging in age from 5 to 28 years, with an average of 12.5 years. The course of disease ranged from 1 to 5 years. The main symptoms included variable mass and pain, partly with repeated burst bleeding. Eighteen patients underwent MRI examination and 11 patients underwent CTA examination. The operative effects were evaluated by the mass changes, pain and recurrence.

RESULTSThere are 6 cases of upper limbs and 12 cases of lower limbs. All pathologic types of all patients with were vascular malformation, in which 13 with blood capillary malformation, 4 with vein malformation, and 1 with arteriovenous fistula malformation. All the patients were followed up, and the duration ranged from 6 months to 2 years,with a mean of 8.8 months. Fifteen patients got an excellent result, 2 good and 1 poor.

CONCLUSIONThe accordance rate of CTA and MRI in the diagnose of intramuscular hemangioma in skeletal muscle are high. CTA 3D stereo-anatomy imaging has a considerable value in choosing the optimal operative methods, protecting important blood vessel and disposing provision blood vessel. As to the preoperative determination of muscle invasive and the decision of operative method, MRI is more valuable than CTA.

Adolescent ; Adult ; Child ; Child, Preschool ; Female ; Hemangioma ; diagnosis ; pathology ; surgery ; Humans ; Magnetic Resonance Imaging ; Male ; Muscle, Skeletal ; pathology ; Tomography, X-Ray Computed