Streptomyces sp. A048 was cultured in a complete medium to the last stage of log phase,the hyphae were washed and collected by centrifugation. Then the hyphae were inoculated in liquid medium for chitinase production using two-step fermentation. Activity of chitinase produced by two-step fermentation was 1.1 times higher than that from one-step fermentation,and ferment cycle was for 54 hours,which was 66 hours shorter than that of one-step fermentation. The hyphae and the powder of chitin were co-immobilizated and cultured in liquid medium for 36 hours,activity of chitinase was 1.8 times higher than that from one-step fermentation,and ferment cycle was 54h shorter than that of one-step fermentation. By adding 0.4% cellulose to two-step fermentation,activity of chitinase was 18.52 U/mL that was 4 times higher than that from the control and 10 times higher than that from one-step fermentation. Two step fermentation with chitin and cellulose may be the optimal fermentation process to produce Chitinase from Streptomyces sp. A048.

0.05). Conclusion There is abnormal thyroid function in patients with anorexia nervosa. Most of the patients with anorexia nervosa are individuals with low T3 syndrome, rather than hypothyroid. Since this syndrome is reversible, thyroxine is not necessary during the treatment.

OBJECTIVETo investigate the influence of hepatitis C virus (HCV) serotype on the interferon (IFN) treatment of patients with chronic hepatitis C.

METHODSNinety-eight patients with chronic hepatitis C were divided into two groups: patients in group 1 (n=53) were treated with Pegasys, 180 ug injected subcutaneously once a week for 24 weeks, and those in group 2 (n=45) were injected with Roferon-A 3 MU three times a week for 24 weeks and then patients in both groups were followed up for another 24 weeks. The virological response at the end of follow up was the primary endpoint for evaluating the effects of IFN treatment. The HCV RNA levels of the chronic hepatitis C patients were determined with COBAS AMPLICOR MONITOR Test, version 2.0, and the HCV serotypes were examined by the means of ELISA using Murex HCV Serotyping 1-6 Assay.

RESULTSOf the 98 cases, HCV in 44 cases was serotype 1, in 23 was serotype 2, in 10 was serotype 3, in 1 was serotype 4, 1 was serotype 5 and in 2 was serotype 6; HCV serotypes in the remaining 17 patients could not determined. In Pegasys treatment group, the biochemical and virological response was not significantly different at the end of treatment between the patients with serotype 1 and non serotype 1 or serotype undetermined patients, but the sustained virological response rate of HCV serotype undetermined group (66.7 percent) was significantly higher than that of serotype 1 patients (27.3 percent) (p=0.035). In Roferon-A treatment group, the virological response rate at 24 weeks and sustained viral response rate at the end of follow-up was not significantly different between serotype 1 and non serotype 1 patients or serotype undetermined patients.

CONCLUSIONAfter the six months treatment course, the HCV serotype had some effects on the treatment response to Pegasys treatment for chronic hepatitic C.

Adult ; Antiviral Agents ; administration & dosage ; Drug Administration Schedule ; Female ; Hepacivirus ; drug effects ; immunology ; Hepatitis C Antibodies ; blood ; Hepatitis C, Chronic ; drug therapy ; immunology ; Humans ; Interferons ; administration & dosage ; Male ; Middle Aged ; Young Adult

Objective To assess the changes on the whole night polysomnography (PSG) in patients with chronic fatigue syndrome(CFS). Methods The whole night PSGs were recorded from 24 patients with CFS and 33 normal subjects. Results Compared with normal subjects, patients with CFS showed significantly reduced total sleep duration ([488.7?21.7]min vs [515.9?31.7]min, P

OBJECTIVETo generate a gene delivery plasmid carrying the dominant negative form of the protein phosphatase 2A catalytic subunit a (DN-PP2Aca) driven by a hepatocellular carcinoma (HCC) tissue-specific promoter and investigate its ability to inhibit growth of cultured hepatoma cells.

METHODSThe gene delivery plasmid was constructed by PCR-amplifying DN-PP2Aca from wild-type PP2Aca using site-directed mutagenesis and then ligating the sequence-verified amplicon downstream of an alpha-fetoprotein enhancer and phosphoglycerate kinase promoter (AFpg) in the luciferase reporter vector pGL3-Basic. Following transfection into two AFP+ hepatoma cell lines (HepG2 and HepG3) and two AFP- hepatoma cell lines (SK-HEP-1 and L02), the transcriptional activity of the AFpg-driven DN-PP2Aca plasmid was tested using luciferase reporter gene assay and western blotting. The effect on cell growth was tested using MTT assay. Between group differences were assessed by t-test.

RESULTSThe AFpg-driven DN-PP2Aca plasmid showed high transcriptional activity and protein expression in both HepG2 and Hep3B cells. At 72 h after transfection, the proliferation capacities were repressed by 42.65%+/-3.99% (P = 0.0002) and 39.87%+/-3.91% (P = 0.0002) in AFP+ HepG2 and Hep3B cells, respectively (vs. untransfected). In contrast, the plasmid was transcriptionally inactive in and had no effect on proliferation of AFP- cells.

CONCLUSIONThe AFpg-driven DN-PP2Aca plasmid exhibits selective cytotoxicity against AFP+ hepatoma cells, and may represent a useful gene therapy strategy to treat HCC.

Carcinoma, Hepatocellular ; genetics ; metabolism ; Enhancer Elements, Genetic ; Genetic Therapy ; Genetic Vectors ; Hep G2 Cells ; Humans ; Liver Neoplasms ; genetics ; metabolism ; Mutation ; Promoter Regions, Genetic ; Protein Phosphatase 2 ; genetics ; alpha-Fetoproteins ; genetics

Aged ; Female ; Humans ; Male ; Middle Aged ; Multiple Myeloma ; diagnosis ; immunology ; Prognosis ; T-Lymphocyte Subsets ; immunology

Objective To observe the effect of ginsenoside Rb1,the most abundant ginsenoside in ginseng root,on differentiation and lipolysis of 3T3-L1 cells and to explore its anti-diabetic mechanism.Methods 3T3-L1 preadipoeytes were induced under standard differentiation process in the presence of 0.1,1,10,100?mol/L ginsenoside Rb_1 for 6 days.Oil red O staining,measurement of triglyceride contents and glucose uptake assay were performed.The expressions of mRNA and protein of PPAR?2,C/EBP?,ap2,glucose transporter (Glut) 1,and Glut4 were analysed with quantitative real time-PCR and Western blot.The binding affinity of Rb_1 to PPAR?-LBD was evaluated by Surface Plasmon Resonance (SPR).Lipolysis of adipocytes was examined by the measurement of glycerol released from adipoeytes treated with Rb_1 for 1 h.Results Ginsenoside Rb_1 facilitated differentiation of 3T3-L1 preadipoeytes in a dose-depondent manner.10?mol/L ginsenoside Rb_1 increased lipid accumulation by about 56%.Treatment of differentiating adipocytes with 10?mol/L ginsenoside Rb_1 increased the expressions of PPAR?2 and C/EBP?mRNA and protein,as well as mRNA expression of ap2,one of their target genes.After treatment of differentiating adipoeytes with Rb_1,basal and insulin-mediated glucose transport augmented significantly accompanied by up-regulations of mRNA and protein level of Glut4,but not of Glutl.SPR showed Rb_1 could bind to PPAR?which suggested Rb_1 was a ligand of PPAR?.Ginsenoside Rb_1 inhibited basal lipolysis in adipoeytos in a dose-dependent manner.However,it did not affect isoproterenol-stimulated lipolysis.Conclusion As a PPAR?ligand,ginsenoside Rb_1 promotes adipogenesis,inhibitas basal lipolysis and inereasos basal and insulin-mediated glucose transport in cultured adipoeytes.Therefore,anti-diabetic and insulin-sensitizing activity of ginsenosides is,at least in part,involved in the enhancing effect on PPAR?2 and C/EBP?expressions,hence promoting adipogenesis and glucose uptake,and inhibiting lipolysis in adipocytes.

OBJECTIVETo evaluate whether the rapid viral response (RVR) to combinational therapy with interferon and rabavirin can be used to predict the sustained viral response (SVR) in chronic hepatitis C patients.

METHODSAccording to their clinical characteristics, all patients in this study were given pegylated or conventional interferon injection and different dose of ribavirin according to their weight. Patients were injected Pegasys (pegierferon alpha-2a) 180 microg or 135 microg once a week, or pegyintron 50-80 microg once a week, or conventional interferon 3-5 MU every two days, in combination with a dose of 600-1500 mg/d ribavirin. The serum HCV RNA load was determined at 0, 4, 12 week, and then every 12 weeks. After the viral response obtained, the patients were treated for another 24-72 weeks and followed up 24 weeks. The main parameter to evaluate the efficacy was SVR rate. The influence factors associated with rapid viral response were investigated.

RESULTSRVR was obtained at week 4 in 84.2% of the 120 patients. The HCV RNA baseline of RVR group was (5.883+/-1.246) lg copies/ml, which was significantly lower than that of the group without RVR [(6.502+/-0.693) lg copies/ml, t=2.15, P=0.034]. 97 patients with RVR who finished treatment and follow-up, 90.7% of these patients obtained SVR, but the SVR rate in patients (82.4%) without RVR was lower than that in patients with RVR (x2=0.371, P=0.543). In this study, RVR rate was not associated with HCV genotype and the dose of interferon used. In the naive patients, the RVR to pegylated interferon was 87.8%, which was significantly higher than that in retreat patients (x2=4.651, P=0.031).

CONCLUSIONHigh RVR rate could be obtained in chronic hepatitis C patients treated combined with interferon and ribavirin. RVR rate is associated with the HCV RNA baseline load in both naive and retreat patients but not correlated to HCV genotype. RVR could predict the SVR.

Administration, Cutaneous ; Adolescent ; Adult ; Aged ; Antiviral Agents ; administration & dosage ; therapeutic use ; Child ; Drug Therapy, Combination ; Female ; Genotype ; Hepacivirus ; drug effects ; genetics ; Hepatitis C, Chronic ; blood ; drug therapy ; pathology ; Humans ; Interferon-alpha ; administration & dosage ; therapeutic use ; Male ; Middle Aged ; Polyethylene Glycols ; administration & dosage ; therapeutic use ; Predictive Value of Tests ; RNA, Viral ; blood ; Recombinant Proteins ; Recurrence ; Ribavirin ; administration & dosage ; therapeutic use ; Time Factors ; Treatment Outcome ; Viral Load ; Young Adult

Adipose tissue is not simply a depot of energy, but is an active endocrine organ. The adipokines play an important role in the pathogenesis of metabolic syndrome. The proinflammatory adipokines secreted from expanded visceral adipose tissue directly induce insulin resistance and vascular injuries. A better understanding of the endocrine function of adipose tissue may lead to more rational therapy for metabolic syndrome.
Adiponectin ; physiology ; Adipose Tissue ; physiopathology ; Drug Design ; Leptin ; physiology ; Metabolic Syndrome ; drug therapy ; physiopathology ; Resistin ; physiology ; Tumor Necrosis Factor-alpha ; physiology

Syringin is one of the main bioactive ingredients in Saussurea involucrata. In this study, various chromatographic techniques were employed to isolate and purify syringin in the polar extraction of cell suspension cultures of S. involucrata. The structure of syringin was characterized by the analysis of spectroscopic data. A quantitative analytical method for the content of syringin in cultures of S. involucrata was established with RP-HPLC. The method is convenient, accurate and reliable. All this results provided a basis for further studies on application of cell suspension cultures of S. involucrata for large-scale production of bioactive compound syringin.
Cell Culture Techniques ; Chromatography, High Pressure Liquid ; methods ; Glucosides ; analysis ; isolation & purification ; Phenylpropionates ; analysis ; isolation & purification ; Plants, Medicinal ; chemistry ; cytology ; Reproducibility of Results ; Saussurea ; chemistry ; cytology