This study investigated the role of glucose in the biogenesis of high-density lipoprotein cholesterol (HDL-C). Mouse primary peritoneal macrophages were harvested and maintained in Dulbecco's modified Eagle's medium (DMEM) containing glucose of various concentrations. The cells were divided into 3 groups in terms of different glucose concentrations in the cultures: Control group (5.6 mmol/L glucose), high glucose concentration groups (16.7 mmol/L and 30 mmol/L glucose). ATP-binding cassette transporter A1 (ABCA1) mRNA expression in the macrophages was detected by semi-quantitative RT-PCR 24, 48 and 72 h after glucose treatment. The results showed that ABCA1 mRNA expression in the 16.7 mmol/L glucose group was not significantly different from that in the control group at all testing time points (P>0.05 for each). In the 30 mmol/L glucose group, macrophage ABCA1 mRNA expression was not changed significantly at 24 h (P=0.14), but was substantially decreased by 40.4% at 48 h (P=0.009) and by 48.1% at 72 h (P=0.015) as compared with that in the control group. It was concluded that ABCA1 is of vital importance for HDL-C biogenesis. High glucose may hamper HDL-C biogenesis by decreasing ABCA1 expression, which contributes to low HDL-C level in diabetes.

Background and purpose:Studies have shown that ribosomal protein L8 (RPL8) is shared by melanomas, gliomas and ovarian carcinomas. A peptide of RPL8 signiifcantly stimulated proliferation and cytokine expression of the hepler T cell clone and lymphocytes in melanoma patients. RPL8 may stimulate anti-tumor immunity, making RPL8 an attractive candidate for therapeutic intervention. In this study, we prepared DC pulsed by RPL8 (RPL8-DC) and investigate the anti-tumor effect of RPL8-DC on melanoma in mice.Methods: The recombinant protein was achieved through IPTG induction in E. coli and identiifed with Western blot. Bone marrow-derived DC was loaded with RPL8 protein. RPL8 and CD11c, CD80, MHC-Ⅰ, MHC-Ⅱmolecules on dentritic cells were monitored by lfuorescence microscope and FACS analysis, respectively. The anti-tumor effect of T cells in vitro was detected by MTT assay. Subcutaneous tumors were induced in C57BL/6 mice using B16 cells. The tumor volumes were measured after injection with RPL8-DC. Results:The puriifed protein was combined with speciifc antibodies. DCs pulsed by RPL8 were visualized under lfuorescent microscopy. CD11c, CD80, MHC-Ⅰ, MHC-Ⅱmolecules on DCs were up-regulated after stimulation with RPL8 and LPS. B16 cells were inhibited by T cells stimulated with RPL8-DC. The inhibition rate of tumor cells was 70%in RPL8-DC group when effector-to-target ratio was 30∶1, which was higher than PBS and DC groups. Inhibition of growth could be observed more signiifcantly in mice after the treatment with RPL8-DC. The mice receiving the therapy of RPL8-DC were able to survive much longer than the mice receiving control therapy. Conclusion:The DC pulsed by RPL8 protein can inhibit the growth of melanoma.

ObjectiveTo evaluate whether AN69 ST membrane would prolong filter lifetime in continuous renal replacement therapy (CRRT) without anticoagulation in patients with high risk of bleeding.Methods A single-center, prospective, randomized, double-blind control trial with crossover design was conducted. From March 1st to December 31st in 2013, patients who were admitted to Department of Critical Care Medicine of the Fourth Hospital of Hebei Medical University meeting CRRT treatment indications, but could not receive systemic anticoagulation because of high risk of bleeding were studied. The selected patients were randomly divided into two groups according to a random number table, and four filters consisting of two AN69 ST100 membrane filters (A) and two traditional AN69 M100 membrane filters (B) were used for them. GroupⅠ with the filter order of A-B-A-B, and groupⅡ with the order of B-A-B-A. The clinical data of patients was recorded in detail, and conventional AN69 ST and AN69 membrane filter lifetime, their influence on coagulability, and the incidence of bleeding complications were compared.Results Seventeen patients were enrolled, with 10 in groupⅠ, and 7 in groupⅡ. The basic medical characteristics including gender, age, acute physiology and chronic health evaluationⅡ (APAECHⅡ) score, sequential organ failure score (SOFA), Acute Renal Injury Network (AKIN) stage, activated partial thromboplastin time (APTT), prothrombin time (PT), international normalized ratio (INR), platelet count (PLT), and use of mechanical ventilation were not significantly different between two groups. But the use of vasoactive drug was more frequent in groupⅡcompared with that of groupⅠ[100.0% (7/7) vs. 30.0% (3/10),χ2 = 8.330,P = 0.010]. AN69 ST filter lifetime (n =34) was (15.92±2.10) hours, there was no statistically significant difference compared with that of AN69 membrane (t = 0.088,P = 0.942), filter lifetime of which (n = 34) was (16.12±1.38) hours. It was also found by Kaplan-Meier survival analysis that there was no significant difference between the two membrane filter lifetime (χ2=1.589,P =0.208). Logistic regression analysis showed that the life of the first filter was not correlated with coagulation indicators, including APTT, PT, INR, and PLT [APTT: odds ratio (OR) = 0.977, 95% confidence interval (95%CI) = 0.892-1.071, P = 0.623; PT:OR = 1.001, 95%CI = 0.901-1.109,P = 0.988; INR:OR = 1.078, 95%CI = 0.348-3.340,P = 0.896;PLT:OR = 0.996, 95%CI = 0.974-1.019,P = 0.735]. The application rate of vasoactive drugs, which was different between two groups for basic medical indications showed no effect on filter life time (OR = 2.541, 95%CI = 0.239-26.955,P = 0.439). Reasons of clotting in filters were also analyzed, and it was found that blood coagulation in the filter ranked the top (88.2%), and the other reasons were catheter-related problems, death, and unscheduled transport. No difference in blood coagulation function was found in both groups after treatment for 12 hours, and there was no bleeding complication.ConclusionDuring the CRRT without systemic anticoagulant, both surface-treatment with polyethyleneimine AN69 and AN69 ST membrane cannot prolong filter lifetime.

Aim To investigate the effects of ghrelin on alcohol-induced liver injury. Methods The alcoholic liver injury mouse model was induced by chronic etha-nol feeding ( 4-week ad libitum oral feeding with the ethanol liquid diet) plus a single binge ethanol (5 g· kg-1 ) feeding. The level of alanine aminotransferase (ALT), aspartate aminotransferase (AST) in serum, malondiadehyde ( MDA ) content, superoxide dis-mutase (SOD) and glutathione peroxidase (GSH-Px) activities in liver homogenate were assayed by spectro-photometer. Hepatic pathological examination was ob-served by HE staining. The mRNA expression of proin-flammatory cytokines including TNF-α, IL-1β, IFN-γ, IL-6 and MCP-1 in the liver was measured by real-time PCR method. Results This chronic-plus-single-binge high dose ethanol feeding synergistically induced liver injury, inflammation and fatty liver change. Treatment with Ghrelin ( 5 , 10 , 20 μg · kg-1 ) significantly de-creased the enhanced level of transaminase ( ALT, AST) in serum, improved the pathologic change in liv-er, and reduced the infiltration of inflammatory cells induced by alcohol administration. Ghrelin also de-creased MDA content and increased the reduced SOD and GSH-Px level in liver homogenate. Furthermore, ghrelin decreased inflammatory cytokines mRNA ex-pression including TNF-α, IL-1β, IFN-γ, IL-6 and MCP-1 in the liver. Conclusion Ghrelin has protec-tive effects against alcoholic liver injury in mice via in-hibiting inflammation and suppressing oxidative stress.

Objective To investigate demographic factors implicated in the functional maturation of autologous arteriovenous fistula in a population of patients with end-stage renal disease. Methods The data of 335 consecutive patients with end-stage renal disease who were performed autologous arteriovenous fistula from January 2010 to December 2015 were analyzed retrospectively. The parameters affecting autologous arteriovenous fistula functional maturation were screened. Results Overall arteriovenous fistula functional maturation rate was 78.14%(218/279). The arteriovenous fistula functional maturation rate was 68.33%(82/120) in diabetes and 85.54%(136/159) in non-diabetes, and there was significant difference (χ2=11.844, P<0.01). The arteriovenous fistula functional maturation rate was 62.26%(33/53) in hypoproteinemia and 81.86%(185/226) in non-hypoproteinemia, and there was significant difference (χ2=9.648, P<0.01). Diabetes and hypoproteinemia were the risk factors to promote functional maturation (OR=6.003, 8.476). The arteriovenous fistula functional maturation rate was 87.10%(81/93) in calcium channel blockers using and 73.66%(137/186) in non-calcium channel blockers using, and there was significant difference (χ2=6.556, P<0.05). Calcium channel blockers was the protective factor for promoting functional maturation (OR=0.086). Conclusions Diabetes and hypoproteinemia are found to be associated with functional non-maturation, while calcium channel-blocker agents are associated with better functional maturation.

AIM To construct a non-normalized cDNA library from Agkistrodon acutus venom gland as an imtial step to develop new and more effective venom by genetic engineering technique for screening and expressing target genes. METHODS The total RNA was extracted from fresh venom gland using Trizol. mRNA was reversely transcripted to cDNA using superscriptⅡ reverse transcriptase. Second-strand synthesis was performed using DNA polymeraseⅠ. After adding EcoRⅠ adaptor, phosphorylating the end and digesting with XhoⅠ, the cDNA was collected in five fractions (<0.25 kb, 0.25-0.5 kb, 0.5-1 kb, 1-2 kb and >2 kb) using the QIAquick Gel Extraction kit and ligated to pBluescriptⅡ vectors. The five libraries obtained were plated by infecting E.coli DH10B, constructing a cDNA library of Agkistrodon acutus venom gland. Sequencing clones at random, 8696 high quality 5′ end expressed sequenced tags (ESTs) were obtained and analyzed. The initial sequences were assembled into 2855 clusters. Among which, one of the clusters (Agkihagin) consisting of 74 ESTs was identified as a novel metalloprtoteinase based on RT-PCR and sequence analysis. RESULTSThe titers of library were 2.048×106. The novel metalloproteinase belonged to PⅢ type metalloproteinase. Its open reading frame was composed of 1827 nucleotides and coded a pre-zymogen of 608 amino acid with zinc-binding domain for metalloproteinase and Asp-Glu-Cys-Asp(DECD) domain for disintegrin. CONCLUSION The capacity of cDNA library of venom gland is above the general level of cDNA library. It would be a helpful platform to construct a catalog for transcripts in the venom gland of the Agkistrodon acutus. The sequence analysis indicates that the deduced amino acid sequence of the identified gene for metalloproteinase share the highest 87% identity with the metalloproteinase genes of other snakes in the GenBank. It lays a good foundation for the study of structure-function relationships of snake venom metalloproteinases.

Objective To assess the mutation in exon 8 of C1 esterase inhibitor(C1INH)gene in a patient with hereditary angioedema(HAE).Methods Genomic DNA was extracted from a female patient with HAE as well as her mother and a normal human control.The fragment of exon 8 of C1INH gene was amplified by PCR and inserted into plasmid carrier pUC19 with the help of ligase.Then,the recombinant plasmid was transformed into competent cells of E coli TG1 strains.After culture of positive transformant,plasmid DNA Was extracted and subjected to sequencing.SDS-PAGE and We:stem blot were performed on the sera of the patient to detect the concentration and function of C1INH protein.Results An A1677G mutation at exon 8 of C1INH gene.which resulted in a substitution of isoleucine to valine at codon 440,Was found in the patient who SUfiered from HAE type I.Additionally.SDS-PAGE and Western blot revealed that the molecular weight of C1INH protein was 96 000.but not 105 000 observed in noHnal human control.Conclusion The newly identified mutation 1440V.which is located at P4 residue of reactive center loop in C1INH.may result in conformational alteration of C1INH.

OBJECTIVETo analysis deacclimatization symptom scores of 159 youth from Karakoram and Tibet Ali area, and provide the basis for the development of relevant prevention and control measures.

METHODSUsing the method of epidemiological symptoms questionnaire, 18 symptoms of 190 youth who returned to the plain area from the different plateau were investigated. The symptom scores of different altitude, age, the time of staying, different units, continuous or intermittent stage and education were surveyed.

RESULTSDeacclimatization symptom scores among 5,000 meter groups were significantly higher than those of 4,300 meter and 3,700 meter group (P < 0.05, P < 0.01). There was no significant difference between the 4,300 meter group and the 3,700 meter group (P > 0.05). There were significant differences among the stayed personnel (different age, position, unit, education, time, continuous or intermittent) (P < 0.01). There was significant difference between the continuous defended the group and intermittent group (P < 0.01).

CONCLUSIONDeacclimatization symptom scores were related to the plateau exposure time, altitude, workload, plateau continued exposure. The older, the longer exposure, the higher altitude, the greater workload at plateau were showed higher deacclimatization symptom score.

The content of benzyl isothiocyanate (BITC) which as the enzymatic hydrolysis product of benzyl glucosinolate through thioglucosidase was determined by HPLC. The content of benzyl isothiocyanate (BITC) which as the enzymatic hydrolysis product of benzyl glucosinolate through thioglucosidase was determined by HPLC. The chromatography condition was as follows: Kaseisorb LC ODS 2000 (4.6 mm x 150 mm, 5 min) column with the mobile phase of acetonitrile(A)-water( B) under gradient elution (0-5 min, 3%-8% A; 5-9 min, 8%-48% A; 9-23 min, 48%-62% A; 23-28 min, 62%-99% A); the flow rate was 1.0 mL x min(-1) with 10 microL injection volume; detection wavelength was 246 nm and temperature of column was 40 degrees C. The content of benzyl glucosinolate was in the range of 10.76-17.91 g x L(-1). The method is simple, accurate and good reproducibility which can be used for the determination of benzyl glucosinolate in Lepidium meyenii, effectively.
Chromatography, High Pressure Liquid ; methods ; Glucosinolates ; analysis ; Lepidium ; chemistry ; Plant Extracts ; analysis

OBJECTIVETo systematically review the efficacy and safety of sodium ferulate (SF) for the treatment of diabetic nephropathy.

METHODSBy computerized retrieving the Cochrane Library, MEDLINE, EMBASE, CNKI, VIP, CBM (theses, conference and internet materials), as well as data from internet materials regarding randomized controlled clinical trials of sodium ferulate for the treatment of diabetic nephropathy were collected completely. Data were strictly extracted using the simple evaluation method recommended in Cochrane Handbook and Meta-analysis was performed using Revman 5.0 software.

RESULTSFourteen randomized controlled trials involving 906 patients met the inclusion criteria. Meta-analysis showed that as compared with the control group, the effects in SF group were superior in terms of reducing urinary albumin excretion rate (UAER) at early stage [WMD = 16.08, 95% confidence interval (95% CI): 11.01 to 21.15] and clinical stage (WMD = 82.66, 95% CI: 66.95 to 98.37), urinary endothelin/endothelin-1 (ET/ET-1, WMD = 10.78, 95% CI: 8.18 to 13.39), levels of serum creatinine (SCr, WMD = 6.42, 95% CI: 1.83 to 11.01), blood urea nitrogen (BUN, SMD = 1.45, 95% CI: 0.19 to 2.71) and total cholesterol (TC, WMD = 0.84, 95% CI: 0.56 to 1.21, as well as in increasing high density lipoprotein-cholesterol (HDL-C, WMD = 0.17, 95% CI: 0.09 to 0.26), showing significant difference between groups. However, the effects of SF were insignificantly different to those of control in reducing fasting blood glucose (FBG, WMD = 0.17, 95% CI: -0.03 to 0.37) and triglyceride (TG, SMD = -0.13, 95% CI -0.49 to 0.23).

CONCLUSIONSAt present the evidences show that SF is superior to the conventional treatment in reducing UAER, ET, SCr, BUN, TC and increasing HDL-C, but there is no evidence to show that SF is superior in reducing FBG and TG. However, the evidence is not strong enough due to the low quality of included literature. More large-scale, multi-center, randomized trials are needed to confirm the efficacy and safety of SF in treating diabetic nephropathy.