OBJECTIVETo observe the effect of pure total flavonoids from Citrus (PTFC) on the hepatic fatty degeneration, inflammation, oxidative stress and SIRT1/PGC-1alpha expressions in mice with non-alcohol steatohepatitis (NASH), and discuss the action mechanism of PTFC on NASH.

METHODMice were given high-fat diet for 16 weeks to induce the NASH model. Since the seventh week after the model establishment, the mice were intervened with 100, 50 and 25 mg x kg(-1) x d(-1) PTFC for 10 weeks. The pathologic changes in hepatic tissues were observed with HE staining. The contents of TG, CHOL in hepatic tissue, as well as the levels of AST, ALT in serum were detected by using the biochemical process. The expression of SIRT1, PGC-1alpha and MnSOD mRNA in hepatic tissues were detected with Real-time PCR assay. SIRT1, PGC-1alpha protein and 8-OHdG expressions were determined with the immunohistochemical method. The SOD level in hepatic tissues was tested by the xanthine oxidase method. The MDA content in hepatic tissues was examined by the thiobarbituric acid method.

RESULTThe contents of TG, CHOL, NAFLD activity scores and ALT level in serum in hepatic tissues of mice in the model induced by fat-rich diet were obviously higher than that of the normal group (P < 0.010. The SIRT1, PGC-1alpha, MnSOD mRNA and protein expression in hepatic tissues were significantly lower than that of the normal group (P < 0.01). The expression of 8-OHdG and the content of MDA in hepatic tissues were obviously higher than that of the normal group (P < 0.01). After the intervention with different doses of PTFC, the NAFLD activity scores, the content of TG and the level of AST in serum were notably lower than that of the normal group (P < 0.01, P < 0.05); whereas the SIRT1, PGC-1alpha, MnSOD mRNA and protein expression were obviously higher than that of the normal group (P < 0.01, P < 0.05), with the significant decrease in the expression of 8-OHdG and the content of MDA (P < 0.01).

CONCLUSIONOxidative stress/lipid peroxidation enhancement in in NASH mice induced by high-fat diet may be related to the changes in SIRT1/PGC-1alpha signal transduction pathway. PTFC could enhance the anti-oxidant capacity in liver, relieve the damage of reactive oxygen during the fatty acid metabolic process, and prevent NASH from the occurrence and development by regulating the SIRT1/PGC-1alpha signal pathway.

Animals ; Citrus ; chemistry ; Fatty Liver ; drug therapy ; genetics ; metabolism ; Flavonoids ; chemistry ; pharmacology ; Inflammation ; drug therapy ; genetics ; metabolism ; Liver ; drug effects ; metabolism ; Male ; Mice ; Mice, Inbred C57BL ; Non-alcoholic Fatty Liver Disease ; Oxidative Stress ; drug effects ; genetics ; Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha ; Sirtuin 1 ; genetics ; metabolism ; Transcription Factors ; genetics ; metabolism

?AlM: To analyze and study the changes of intraocular pressure ( lOP) , visual field and P-ERG on patients with big cup/disk (C/D) in 24h.
?METHODS: A total of 120 cases ( 240 eyes ) diagnosed with big C/D (C/D>0. 3) were divided into group A (C/D<0. 6, 67 cases, 114 eyes ) and group B ( C/D≥0. 6, 73 cases, 126 eyes). Forty cases (80 eyes) with small C/D (C/D≤0. 3) were chosen as control group. All cases underwent 24h lOP examination, vision examination of 30-2 SlTA - standard static threshold and pattern electroretinogram ( P-ERG) examination. The differences between the examination indexes of the three groups were analyzed.
?RESULTS:There was no significant statistical difference in the 24h average lOP (P>0. 05) among the three groups, while the amplitudes had significant statistical differences (P<0. 05). Compared with the control group, the lOP amplitudes of group B were obviously higher, and the difference was of statistical significance (P<0. 05). ln terms of static threshold visual fields, the mean deviations (MD) and pattern standard deviations (PSD) of the three groups had significant statistical differences ( P < 0. 05 ). Comparison between every two groups:compared with the control group, the MD and the PSD were significantly increased in group B, and the difference was of statistical significance (P<0. 05); while there was no significant statistical difference in group A (P>0. 05);Compared with group A, the MD and the PSD were significantly higher in group B, and the difference was of statistical significance (P<0. 05). ln terms of P-ERG examination, there was no significant statistical difference in P50’s latent periods among the three groups (P>0. 05), but there were significant statistical differences in amplitudes (P<0. 05). Comparison between every two groups: compared with the control group, the P50’s amplitude was significantly decreased in group B, and the difference was of statistical significance (P<0. 05), while there was no significant statistical difference in group A (P>0. 05); Compared with group A, the P50’s amplitude of group B was significantly lower, and the difference was of statistical significance (P<0. 05).
?CONCLUSlON: When the patients’ C/D is no less than 0. 6, there are obvious changes of 24h lOP, static threshold visual field and P-ERG’s P50 wave. For patients with big C/D, a single lOP examination is far from sufficient, so an in-depth visual function examination should be performed. Meanwhile, it’s feasible to regard C/D 0. 6 as a screening criterion for suspected glaucoma.

OBJECTIVETo identify the expression characteristics of the 1700001022RIK (RIKEN cDNA 1700001022) gene in mice and explore its function by bioinformatic analysis.

METHODSUsing the expression profile of gene microarray, we detected the expression of a new testis-specific gene, 1700001022RIK, in mice. We analyzed its expression characteristics in the testis tissue and their changes in different developmental stages of the testis by RT-PCR, real-time RT-PCR, Western blot, and immunohistochemistry. We performed bioinformatic analysis using a bioinformatic software.

RESULTSThe 1700001022RIK gene was specifically expressed in the mouse testis in an age-dependent manner, most highly in the adult mice. The 1700001022RIK protein was mainly expressed in the spermatogonia, spermatocytes, and round spermatids of the adult mice. Bioinformatic analysis showed that the 1700001022RIK protein amino acid sequence had a high similarity in human and mice, which indicated that this gene was highly conserved in mammals.

CONCLUSION1700001022RIK is a testis-specific gene mainly expressed in the spermatogonia, spermatocytes, and round spermatids of seminiferous tubules, which might be involved in the regulation of spermatogenesis.

Age Factors ; Animals ; Blotting, Western ; Computational Biology ; DNA, Complementary ; Gene Expression ; Genomics ; Male ; Mice ; Molecular Chaperones ; genetics ; Seminiferous Tubules ; Spermatids ; Spermatocytes ; Spermatogenesis ; genetics ; Spermatogonia ; Testis

Objective To study the exact function of human gene 5 transactivated by pre-S1 protein of hepatitis B virus(PS1TP5)by investigating the gene expression of PS1TP5 in yeast cells. Methods Reverse transcription-polymerase chain reaction(RT-PCR)was performed to amplify the gene of PS1TP5 using the mRNA of HepG2 cells as template and the gene was cloned into pGEM-T vector.The gene of PS1TP5 was cut from pGEM-T-PS1TP5 vector and cloned into yeast expressive plasmid pGBKT7,then pGBKT7-PS1TP5 was transformed into yeast cell AH109.The yeast protein was isolated and analyzed with sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE) and Western hybridization.Results PS1TP5 gene was successfully amplified and identified by DNA sequencing.The digested fragment was cloned into pGBKT7 vector and transformed into yeast cell AH109.The results of SDS-PAGE and Western assay showed that the relative molecular weight of the expressed product was about 36 950,and PS1TP5 protein existed in yeast cells.Conclusion The findings suggest that PS1TP5 can be successfully expressed in yeast cell.

OBJECTIVETo investigate the inhibitory activities of caffeoyl glucopyranoses purified from Balanophora japonica Makino on HIV entry and their mechanism.

METHODSHIV-1 Env pseudovirus was used to evaluate the anti-HIV-1 activity of those compounds. ELISA and molecular docking were used to study the mechanism of the actions of the active compounds.

RESULTSWe used the HIV-1 Env pseudovirus to test the anti-HIV-1 activity of the six phenolic compounds (final concentration 25 microg/ml), and found that only 1,2,6-Tri-O-caffeoyl-beta-D-glucopyranose (TCGP) and 1,3-Di-O-caffeoyl-4-O-galloyl-beta-D- glucopyranose (DCGGP) could effectively inhibit the entry of HIV-1 Env pseudovirus into the target cells in a dose-dependent manner, with IC(50) values of 5.5-/+0.2 and 5.3-/+0.1 microg/ml, respectively. These two compounds could also blocked the gp41 six-helix bundle formation. Molecular docking analysis suggested that they might bind to the hydrophobic cavity of the gp41 N-trimeric coiled-coil.

CONCLUSIONTCGP and DCGGP are potent HIV-1 entry inhibitors targeting gp41 and can serve as lead compounds for developing novel anti-HIV-1 microbicides for prevention of sexual HIV-1 transmission.

Anti-HIV Agents ; pharmacology ; Balanophoraceae ; chemistry ; Cell Line ; Gallic Acid ; analogs & derivatives ; pharmacology ; Glucose ; analogs & derivatives ; pharmacology ; HIV-1 ; drug effects ; Humans ; Hydrolyzable Tannins ; pharmacology ; Plant Extracts ; pharmacology

OBJECTIVETo investigate the changes of blood corpuscles of patients with severe acute respiratory syndrome (SARS) in Beijing.

METHODSTotally 43 patients (21 male and 22 female, 19-74 years old age range) diagnosed as of probable SARS were included in this study. Their corpuscles in the peripheral blood were tested every two days, and the results were analyzed.

RESULTSPatients with SARS were more likely to develop leukocytopenia, lymphopenia and thrombocytopenia in the early period of disease than those in control group. The situation, especially lymphopenia and thrombocytopenia, could not be reversed in patients who died. Persistent low counts of lymphocytes and platelets at presentation might be associated with adverse outcomes.

CONCLUSIONLow counts of leukocytes, lymphocytes and platelets were common among patients in the early stage of SARS. Persistent Lymphopenia and thrombocytopenia may be associated with the prognosis.

Adult ; Aged ; Early Diagnosis ; Female ; Humans ; Leukocyte Count ; Leukocytes ; cytology ; Male ; Middle Aged ; Prognosis ; Severe Acute Respiratory Syndrome ; diagnosis

Based on the full-length cDNA of BmalphaTX14 from Chinese scorpion Buthus martensii Karsch (BmK), gene of the mature peptide of BmalphaTX14 was cloned into the yeast expression vector pPIC9K. After transforming, screening and inducing, tricine-SDS-PAGE and Western blot proved that rBmalphaTX14 protein was expressed in the medium for up to 84 hours, getting nearly 120 mg/L. Recombinant BmalphaTX14 was purified rapidly and efficiently through Ni-NTA-agarose, polyethylene glycol precipitation and gel filtration chromatography. The purified rBmalphaTX14 proved to have the anti-insect activity by toxicity assay. Meanwhile, genomic gene of BmalphaTX14 was cloned and sequenced by PCR method, sequence analysis of this gene showed that BmalphaTX14 had an intron of 408 base pairs located at the signal peptide encoding region, which was similar with the characteristic of other alpha-type sodium ion-channel toxin. Considering both the genomic organization and the peptide function, BmaTX14 proved to be a membership belonging to alpha-type sodium ion-channel toxin.
Amino Acid Sequence ; Animals ; Base Sequence ; Cloning, Molecular ; Molecular Sequence Data ; Pichia ; genetics ; metabolism ; Recombinant Proteins ; biosynthesis ; genetics ; isolation & purification ; Scorpion Venoms ; biosynthesis ; genetics ; Scorpions ; genetics ; Sequence Analysis

OBJECTIVETo study the prognosis and the factors affecting the prognosis in children with acute respiratory distress syndrome (ARDS).

METHODSSeventy-eight children with ARDS were enrolled. The states of their survival within 30 days were followed-up.

RESULTSOf the 78 children with ARDS, 51 cases demised, 27 cases survived, with a 30-days survival rate of 35%. The average survival time was 14.4 days (median: 8 days). The peak of death appeared within 3 days after ARDS. There were significant differences in aspects of age, primary disease, percentage of neonatal hyaline membrane disease, pediatric critical illness score (PCIS), duration of mechanical ventilation, oxygenation index (PaO(2)/FiO(2)), white blood cell count and number of involved organs between the died and survived children (P<0.05 or 0.01). The Cox multiple factors analysis showed that the age (HR 3.924~3.938), primary disease (HR=1.817) and PCIS (HR=0.469) were the risk factors of death.

CONCLUSIONSThe peak of death usually appears within 3 days after ARDS. Age, primary disease and PCIS are the independent factors of prognosis in children with ARDS.

Adolescent ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Infant, Newborn ; Male ; Multivariate Analysis ; Prognosis ; Respiratory Distress Syndrome, Adult ; mortality ; Survival Rate

OBJECTIVETo investigate the expression characteristics of the gene of coiled-coil domain-containing protein 70 (Ccdc70) in the mouse testis and its potential role in spermatogenesis.

METHODSUsing expression profile microarray, we screened the mouse testis-specific gene Ccdc70, studied its expression characteristics in the mouse testis by RT-PCR, real-time PCR, Western blot and immunohistochemistry, followed by bioinformatic analysis of the Ccdc70 protein.

RESULTSThe Ccdc70 gene was expressed highly in the testis but lowly in the epididymis of the mice. The Ccdc70 protein was expressed mainly in the spermatocytes and round spermatids of the testis and in the epithelial cells of the epididymis. Bioinformatic analysis showed a structural domain in the Ccdc70 protein, which was highly conserved in mammalian evolution.

CONCLUSIONThe Ccdc70 gene is highly expressed in the mouse testis and mainly in the spermatocytes, round spermatids, and epididymal epithelial cells, which indicates that it is involved in the regulation of spermatogenesis and epididymal sperm maturation.

Animals ; Computational Biology ; Gene Expression Regulation, Developmental ; Male ; Mice ; Proteins ; genetics ; Spermatogenesis ; genetics ; Testis ; metabolism

OBJECTIVETo explore whether the inhibitory effect of Genipin on uncoupling protein-2 (UCP-2) in mitochondria is involved in energy metabolism of androgen-independent PC3 prostate cancer cells.

METHODSPC3 prostate cancer cells were cultured and treated with Genipin at the concentrations of 40, 80, and 160 μmol/L for 48 hours. Then the proliferation of the cells was detected by MTT assay, the expression of UCP-2 mRNA determined by RT-PCR, and the content of intracellular pyruvic acid (PA) and the activity of succinate dehydrogenase (SDH) in the mitochondria measured by visible spectrophotometry.

RESULTSWith the increased concentration of Genipin, the proliferative activity of the PC-3 cells, the expression level of UCP-2 mRNA, the content of intracellular PA and the activity of SDH in the cells were all decreased, namely, with the enhanced inhibitory effect of Genipin on UCP-2, a trend of reduction was observed in the proliferation of the cells, intracellular PA content, and SDH activity in the mitochondria.

CONCLUSIONGenipin is involved in the energy metabolism of androgen-independent PC3 prostate cancer cells by reducing the content of intracellular PA and the activity of SDH in the mitochondria, which may be associated with its inhibitory effect on UCP-2.

Cell Line, Tumor ; drug effects ; Energy Metabolism ; Humans ; Ion Channels ; metabolism ; Iridoids ; pharmacology ; Male ; Mitochondria ; metabolism ; Mitochondrial Proteins ; metabolism ; Prostatic Neoplasms ; metabolism ; Pyruvic Acid ; metabolism ; RNA, Messenger ; Succinate Dehydrogenase ; metabolism ; Uncoupling Protein 2