A new attempt with lipoprotein lipase agonists in the treatment of nonalcoholic steatohepatitis / 中华肝脏病杂志

Objective@#To analyze non-alcoholic steatohepatitis (NASH)-related differentially expressed genes (DEGs) by bioinformatics methods to find key pathways and potential therapeutic targets for NASH.@*Methods@#GSE61260 chip was downloaded from the public microarray database and liver biopsy samples from 24 NASH cases and 38 healthy controls were included. The Limma software package in R language was used to screen DEGs under the condition of difference multiple > 1.5 and adj. P < 0.05. The clusterProfiler software package was used for GO analysis and KEGG analysis. The STRING online database was used for protein-protein interaction analysis, and the L1000 and DrugBank databases were used for drug prediction.@*Results@#Compared with healthy control group, 857 DEGs were screened out in NASH group including 167 up-regulated genes and 690 down-regulated genes. GO analysis showed that DEGs were mainly involved in inflammation and cholesterol metabolism. KEGG analysis showed that DEGs were mainly enriched in PPAR, non-alcoholic fatty liver disease, oxidative phosphorylation and other signaling pathways. Among them, eight genes of ACSL4, CYP7A1, FABP4, FABP5, lipoprotein lipase, ME1, OLR1 and PLIN1 were enriched in PPAR signaling pathway, and 165 interaction nodes were formed with 47 DEGs-encoded proteins. Lipoprotein lipase interacted with 21 DEGs, and its up-regulated expression had improved lipid metabolism, insulin resistance and anti-inflammatory effects. Four drugs (gemfibrozil, bezafibrate, omega-3 carboxylic acid and glycyrrhizic acid) were screened by L1000 and DrugBank to activate lipoprotein lipase. Presently, these four drugs are clinically used to treat hypertriglyceridemia or to improve inflammation. In this regard, we speculated that the pharmacological effects of these four drugs had improved NASH by activating lipoprotein lipase to promote liver lipid metabolism and alleviate inflammation.@*Conclusion@#PPAR signaling pathway is closely associated to the occurrence and development of NASH, and thereby lipoprotein lipase agonist is a new attempt to treat NASH.

Effect of 60Co-γ Irradiation on the Stability of Isoniazid / 中国药师

Objective:To establish an HPLC method for the determination of isoniazid and explore the impact of 60Co-γ on the sta-bility of isoniazid. Methods:A Diamosil C18(2) column(250 mm×4.6 mm,5 μm) was used. The mobile phase consisted of metha-nol and 0.2% phosphoric acid(2 :98). The flow rate was 1.0 ml·min-1,and the detection wavelength was at 254 nm. The isonia-zid solutions at different accurate initial concentration(10,50,100 μg·ml-1,100 μg·ml-1+saturated N2,100 μg·ml-1+satu-rated O2,100 μg·ml-1+10% tert-butanol and isoniazid powder) were irradiated by 60Co-γ at different doses (0.3,0.6,1,2.5, 5,10 kGy). And then,the concentrations of remained isoniazid were determined by HPLC. Results: Isoniazid was in a good linear relationship within the range of 1-200 μg·ml-1(r=0.999 0) with the recoveriy of 98.13% (RSD=0.65%). The degradation rate of isoniazid was affected by the initial concentration and the radiation dose,which increased with the increase of absorption amount and increased with the concentration increase,and the powder almost had no degradation. The degradation of isoniazid+O2significantly de-creased,and that of isoniazid+N2and isoniazid+tert-butanol was inhibied to a certain extent. Conclusion:Isoniazid is sensitive to 60 Co-γ irradiation,and its degradation is higher with lower concentration and higher irradiation dose.

Role of CD36 in nonalcoholic fatty liver disease / 中华肝脏病杂志

Nonalcoholic fatty liver disease (NAFLD) includes nonalcoholic simple fatty liver, nonalcoholic steatohepatitis, liver cirrhosis, and liver cancer and is the most common liver disease in the world. The complex pathogenesis of NAFLD is a major concern among researchers. CD36 is a transmembrane glycoprotein that takes up fatty acid and binds to oxidized low-density lipoprotein in the liver, and therefore, it is involved in the development and progression of NAFLD. With reference to the latest research findings, this article reviews the association between CD36 and NAFLD and the role of CD36.

The role of cytochrome P450 in nonalcoholic fatty liver induced by high-fat diet: a gene expression profile analysis / 中华肝脏病杂志

Objective@#To clarify the role of cytochrome P450 in nonalcoholic fatty liver disease (NAFLD) by RNA-Seq and bioinformatics analysis.@*Methods@#A total of 20 male C57BL/6 mice were used. Ten mice were fed with high-fat diet (D12492, 60% kcal fat) for 16 weeks to establish a mouse model of NAFLD, and the other 10 mice were fed with low-fat diet (D12450B, 10% kcal fat) as control group. At the end of the experiment, the body weight, liver weight, and hepatic triglyceride (TG) content were measured. Meanwhile, HE staining and RNA-Seq analysis were performed for the liver tissues. The differentially expressed genes were screened out and subjected to bioinformatics analysis, including KEGG and GO BP enrichment analyses and interaction network analysis. Comparison of means between the two groups was made using t-test.@*Results@#Compared with the control group, the mice in the model group were obviously obese, with significantly increased body weight (41.41 ± 6.01 g vs 28.78 ± 1.79 g, t = 6.04, P < 0.01) and liver weight (1.38 ± 0.30 g vs 1.08 ± 0.10 g, t = 2.89, P < 0.01). The mice in the model group showed obvious steatosis, accompanied by a small amount of inflammatory cell infiltration, but with no obvious fibrosis, according to the results of HE staining. In addition, the hepatic TG content in the model group was significantly increased compared with that in the control group (0.64 ± 0.01 mg/mg vs 0.29 ± 0.06 mg/mg, t = 10.11, P = 0.04). Compared with the control group, a total of 367 differentially expressed genes, including 211 down-regulated and 156 up-regulated ones, were identified in the model group according to the RNA-seq results. Meanwhile, 19 CYP450 subtypes, accounting for 5% of the differentially expressed genes, were identified, and CYP2E1, CYP2C70, CYP3A11, CYP3A25, CYP2D26, CYP4A10, CYP17A1, CYP2B10, and CYP2C38 were involved in oxidative stress, steroid hormone metabolism, fatty acid metabolism, arachidonic acid metabolism, and the PPAR signaling pathway. An interaction network was constructed with 30 nodes, and CYP2E1 and CYP2C70 were identified as key nodes. RT-PCR validation results showed that the expression changes of CYP450 subtypes and lipid metabolism-related genes were consistent with the findings of sequencing.@*Conclusion@#The CYP450 family plays a vital role in the pathogenesis of fatty liver by regulating lipid metabolism-related pathways, including oxidative stress, arachidonic acid metabolism, steroid hormone metabolism , and fatty acid metabolism.

Fabrication of the anti-tuberculosis controlled drug delivery system with Ti-PDA-PEG-PLGA-INH and investigation of the biological characteristics / 中华骨科杂志

Objective To fabricate an anti?tuberculosis controlled drug release coating with Ti?PDA?PEG?PLGA?INH and to investigate its surface characteristics, in vivo and in vitro drug release behavior, and tissue biocompatibility. Methods 4?arm?polyethylene glycol (PEG) was synthesized first. Then cover the surface of titanium (Ti) with a layer of poly dopamine (PDA) by Michael addition reaction. Use porous starch and 4?arm?PEG as a carrier, load with isoniazid (INH), then attach to the surface of titanium by casting or sol?gel dip coating methods, and then cover with a layer of poly lactic?co?glycolic acid (PLGA) by the same method, to fabricate the Ti?PDA?PEG?PLGA?INH composite coating finally. The functional group of 4?arm?PEG was charac?terized by proton nuclear resonance spectroscopy (HNMR). The surface characteristics of Ti?PDA?PEG?PLGA?INH were evaluated by scanning electron microscope (SEM), while drug release behaviors were detected by high performance liquid chromatography (HPLC) and the cumulative release rate was calculated, and carry out the antibacterial performance in vitro. The animal model of femoral condyle bone defect was established in 25 New Zealand white rabbits. Titanium rods covered with PDA?PEG?PLGA?INH coating were implanted into defect area. INH concentrations were detected by HPLC in venous blood, muscle and bone tissue at each time point postoperatively. Another 12 rabbits were randomly divided into experimental group and control group, the experi?mental group was implanted with titanium tablets and titanium rods coated with PDA?PEG?PLGA?INH in the paraspinous muscle and left femoral condyles respectively, while the control group was implanted with a blank sheet of titanium tablets and titanium rods in the same place. Hematoxylin and Eosin Staining were used to observe the biocompatibility of the composite system in vivo at 28 and 56 days postoperatively. Results Ti?PDA?PEG?PLGA?INH controlled drug release coating uniformly distributed on the surface of plates and rods, with translucent form and smooth surface. In vitro INH release kinetics exhibited a short?burst release during the first 8h, and the cumulative release of the INH was about 65%. On the 9th day, the cumulative release of the INH was about 90%, and then the release tended to be flat, and the drug release behavior in vitro continued more than 20d. In vivo release test showed that the concentration of INH in vein blood, muscle and bone tissue around the composite system was increased steadi?ly postoperatively. On about the 28th day, the concentration reached the max. However, the INH concentrations in muscle and bone tissue around the composite system were still higher than the minimum inhibitory concentration (MIC) on the 56th day. The antibacterial test in vitro showed that the titanium tablets coated with PDA?PEG?PLGA?INH formed obvious bacterial inhibition zones. The pathological results indicated that mild inflammatory reaction was seen in the 4th week postoperatively, and the reac?tive capsule formed with loose connective tissue. In the 8th week postoperatively, there's no obvious inflammation occurred, and the reactive capsule became more dense and thicker. Conclusion The study successfully fabricated the Ti?PDA?PEG?PLGA?INH anti?tuberculosis controlled drug release coating, with reasonable release behavior both in vivo and in vitro, effective antibac?terial effect of Mycobacterium tuberculosis in vitro and good tissue biocompatibility, which is a potentially effective drug delivery system for spinal tuberculosis.

The amphiphilic curcumin derivative attenuates liver fibrosis by anti-inflammatory and antioxidant effect / 中国药理学通报

Aim To investigate the effects of anti-in-flammation and antioxidation of an amphiphilic curcu-min derivative (Curc-OEG)on CCl4-induced hepatic fibrosis in rats.Methods Rats were randomly divided into four groups:control group,model group,curcumin and Curc-OEG treatment group.All rats except those in control group were given subcuta-neous injection of CCl4 and olive oil mixture,twice a week for 8 weeks.After 4 weeks,rats of control and model group were trea-ted with normal saline intravenously,curcumin group were ad-ministered with curcumin 400 mg.kg -1 .d -1 by gavage and Curc-OEG group were treated with Curc-OEG 1 00 mg.kg -1 .d -1 intra-venously respectively.After 4 weeks treatment,the serum levels of ALT and AST were tested.HE and Sirus staining were used to evaluate the extent of liver inflammation and fibrosis.The mRNA expression levels of proinflammatory cytokines of NF-kB,IL-1 β, IL-6,TNF-α,COX-2 were observed with Real Time PCR.The level of MOD,SOD and GSH in liver of rats were quantified. Results The levels of ALT in control,model,curcumin and Curc-OEG group was (31 .7 ±8.7)U·L -1 ,(383.0 ±75.6) U·L -1 ,(406.3 ±204.7)U·L -1 ,(1 07.0 ±73.7)U·L -1 respectively;that of AST was (1 37.7 ±32.7)U·L -1 ,(585.3 ±36.7)U·L -1 ,(485.0 ±246.5)U·L -1 ,(202.7 ±56.0) U·L -1 respectively,Curc-OEG possessed more hepatoprotective effects than that of curcumin.Liver pathology showed Curc-OEG treatment could significantly alleviate steatosis,reduce inflamma-tion and apparently suppress hepatic fibrogenesis by reducing the thickness of bridging fibrotic septa.Compared with curcumin, Curc-OEG down-regulated mRNA and protein expression levels of NF-kB,IL-1 β,IL-6,TNF-α,COX-2 (P <0.05 ).Moreo-ver,Curc-OEG reduced the level of MOD and increased the lev-els of SOD and GSH.Conclusion Curc-OEG could more sig-nificantly protect the rat liver from CCl4-caused fibrogenesis by anti-inflammatory and antioxidant effect than curcumin.

The study of heat shock protein 70 as a adjuvant carrier on HBV-specific immune response in vitro / 重庆医学

Objective To investigate the effect of mycobacterium tuberculosis heat shock protein 70(TB .HSP70) as an adjuvant carrier on stimulating hepatitis B virus (HBV) core antigen(HBcAg)specific immune response to an accompanying cytotoxic T lym-phocytes epitope peptide from HBV core antigen in vitro .Methods Recombinant proteins HSP70(P1)、HSP70-HBcAg(18-27) (P2)、HSP70-PreS2B (18-24)-PreS2Th(37-53)-HBcAg(18-27)(P3) were expressed in methylotropic yeast Pichia pastoris GS115 . The expression of recombinant proteins was identified by SDS-PAGE and Western blot .The effect of recombinant proteins on den-dritic cell and lymphocytes of chronic HBV infection volunteers was investigated in vitro .The maturation of dendritic cell was meas-ured by flow cytometry ;the secretion of Th1 cytokines such as IL-12p70 ,IL-1β,TNF-αand IFN-γ was measured by ELISA ;the proliferation of lymphocytes was measured by TdR-3H ;the HBV-spesific cytotoxic activity was measured by the classic 51 Cr .Re-sults The recombinant proteins (P1 ,P2 ,P3) were constructed successfully .P1 ,P2 ,P3 could activate dendritic cell from chronic HBV infection volunteers by upregulation CD1a ,CD40 ,CD86 and production Th1 cytokines such as IL-12p70 ,IL-1β and TNF-α. Especially P3 could better induce autologous T cells to generate HBV specific cytotoxic T lymphocytes response ,activate the prolif-eration of lymphocytes and release IFN-γeffectively .However ,the recombinant HSP70 showed no target cell killing and could not induce immune response effectively .Conclusion TB .HSP70 can be used as an adjuvant carrier to stimulate HBV specific immune response to an accompanying cytotoxic T lymphocytes epitope peptide from HBV core antigen ,and enhance immunogenicity of the cytotoxic T lymphocytes epitope peptide .The P3 with B-and T-epitope can activate the HBV specific immune response effectively .

Recent advances in curcumin and its derivatives for treatment of liver diseases / 药学学报

Curcumin is a principal polyphenolic curcuminoid extracted from turmeric rhizome, which has been used for treating inflammation of joints, ulcers, jaundice and other disorders in Asian traditional medicine. In recent years, many studies have indicated that curcumin plays important roles in treatment of liver diseases. Curcumin attenuates liver injury and non-alcoholic fatty liver disease by lowering the release of inflammation cytokines, minimizing oxidative stress, enhancing the sensitivity of insulin and altering lipid metabolism. Curcumin shows potent anti-fibrosis activity, contributing to inhibit the activation of hepatic stellate cells and reduce the deposition of extracellular matrix by its regulation of PPAR-γ, NF-ΚB and TGF-β signaling pathways. Moreover, curcumin exhibits anti-cancer effect by inducing G2/M phase cell cycle arrest and apoptosis in several hepatoma cell lines. However, poor water solubility and low bioavailability of curcumin limit its clinical applications. To overcome its limited systemic bioavailability, many new approaches have been explored to deliver curcumin effectively. This article focuses on advances in the effects of curcumin and its derivatives for treatment of liver injury, non-alcoholic fatty liver disease, liver fibrosis and hepatocarcinoma.

Simultaneous Determination of Chloroginic Acid, Baicalin and Luteolin in Julan Ganmao Particles / 中国药师

Objective:To establish an HPLC method for the simultaneous determination of chloroginic acid, baicalin and luteolin in Julan Ganmao particles. Methods:A Diamosil C18 (2) column(250 mm × 4. 6 mm,5μm) was used. The mobile phase consisted of acetonitrile and 0. 2% phosphoric acid with gradient elution. The flow rate was 1. 0 ml·min-1 , the column temperature was 35℃, and the detection wavelength was at 327 nm for chloroginic acid and baicalin, and 360 nm for luteolin. Results:Chloroginic acid, ba-icalin and luteolin was in a good linear relationship within the range of 0. 50-9. 96 μg·ml-1(r=0. 999 8), 10. 10-202. 00 μg·ml-1 (r=0.999 5), and 0.10-2.04 μg·ml-1(r=0.999 6) with the recovery of 98.13%(RSD=0.65%), 98.80%(RSD=0.26%) and 99. 62%(RSD=0. 57%), respectively. Conclusion: The developed method is simple, accurate and reliable with good repeat-ability, and can be used in the quality control of chloroginic acid, baicalin and luteolin in Julan Ganmao particles.

Advance in the study of poly(lactide-co-glycolide) nano/microparticles as gene vector / 药学学报

Biodegradable nano/microparticles of poly(D, L-lactide-co-glycolide) (PLGA) is a novel non-viral gene vector, which has many advantages, such as safety, non-immunogenicity, easy of large-scale preparation and well load-capability. Therefore, more and more attentions and researches have been paid on its application. Especially, PLGA has shown enormous potential application value and space in the field of plasmid DNA (pDNA) delivery system. On the basis of the current situation of PLGA nano/microparticles for pDNA delivery, this paper focused on summarizing the current preparation approaches and surface modified methods of PLGA particle, furthermore showing its application in gene therapy and genetic vaccine delivery. These showed that PLGA nano/microparticles have extensive prospect in the development of controlled gene delivery system.

Inducement of specific anti-hepatoma immunity of DC activated by HSP70-H22 peptide complexes and sCD40L / 第三军医大学学报

Objective To study the specific anti-tumor immunity of dendritic cell(DC) modified by HSP70-tumor peptide complexes and sCD40L in vitro and in vivo.Methods The murine marrow cells were cultured for 7 d,and then randomized to 4 groups: control,only HSP70-H22 peptide complexes,sCD40L,and HSP70-H22 peptide complexes combined with sCD40L.After intervention for 24 h,IL-12 and IL-10 in the supernatant were detected by ELISA assay,CD40 and CD80 of DC by FACS,and the proliferation of spleen lymphocytes by MRL.The ability of spleen lymphocytes activated by DC to H22 cells in vitro was investigated by MTT.The models of murine hepatoma were established,and then randomized to 5 groups(Ⅰ,Ⅱ,Ⅲ,Ⅳ and Ⅴ),followed by interference with normal saline and DC respectively.At 21 d,mice were sacrificed and the weight of tumor was measured.The levels of IL-10 and IFN-? in blood serum were detected by ELISA assay.Results Compared with that in the groups of control,only sCD40L,and HSP70-H22 peptide complexes,the level of IL-10 in the group stimulated by HSP70-H22 peptide complexes combined with sCD40L decreased significantly,but other indexes in this group increased significantly(P

Relationship between the different replication status of HBV and mutations in core promoter in mother and children infected by mother-to-infant transmission / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To understand the relationship between the different replication status of HBV and mutations in core promoter in mother and child infected by mother-to-infant transmission.</p><p><b>METHODS</b>Core promoter was amplified by PCR and cloned into pGEM-T vector with T-A cloning technique. The recombinant plasmid pGEM-PreS/S was confirmed by digestion with restriction enzyme Apa I and Sac I. Two clones were selected to sequence each patient.</p><p><b>RESULTS</b>Every pair of mother and child had same serotype and genotype and the homology of nucleotides encoding "a" determinant was 98 to 100%. The number of mutations in core promoter in patients with high replication status was more than that of low replication status. Mutations were distributed in BCP and Kunitz-type serine protease inhibitor region mainly. This difference was not associated with mother or child.</p><p><b>CONCLUSIONS</b>The different replication status of HBV is caused by mutations in core promoter in mother and child infected by mother-to-infant transmission and seems not to be associated with the development status.</p>