Syringin, chlorogenic acid and 1,5-dicaffeoylquinic acid are three main bioactive ingredients in herbs of Saussurea involucrata with various pharmacological properties, while their contents are very low. In this study, the biosynthesis of syringin, chlorogenic acid and 1,5-dicaffeoylquinic acid in the cell suspension cultures of S. involucrata were regulated by feeding carbon sources and precursors, which resulted in a great increase of the contents and yields of the above three bioactive ingredients. After 16 days of fermentation, the yields of syringin, chlorogenic acid and 1,5-dicaffeoylquinic acid reached 339.0, 225.3, 512.7 mg x L(-1), respectively. Meanwhile, their contents increased up to 67.9, 1.9, 10.6 times of wild medicinal material, respectively. The results provided a solid basis for further studies on application of cell suspension cultures of S. involucrata for large-scale production of bioactive compounds syringin, chlorogenic acid and 1,5-dicaffeoylquinic acid.
Cell Culture Techniques ; Cells, Cultured ; Chlorogenic Acid ; analysis ; metabolism ; Cinnamates ; analysis ; metabolism ; Glucosides ; analysis ; biosynthesis ; Phenylpropionates ; analysis ; Saussurea ; chemistry ; growth & development ; metabolism

Factors influencing the temperature-sensing accuracy of an ablation catheter are analyzed, in this paper, from the two aspects of the thermocouple temperature sensor, which are the TC length and the TC hole's diameter of the ablation electrode. Meanwhile, differences between products from different companies are given too.
Catheter Ablation ; methods ; Electrodes ; Temperature

To investigate the secondary metabolites of endophytic fungi Pericinia sp. F-31. Column chromatography on silica gel, Sephadex LH-20 and semi-preparative HPLC were used to separate and purify the compounds. Two compounds were isolated from the fermentation broth of Periconia sp. Their structures were identified as 5-(1-hydroxyhexyl) -6-methyl-2H-pyran-2-one (1) and 2-(3-hydroxy-4-methylphenyl) -propanoic acid (2). Compound 1 was a new lactone compound, compound 2 was new natural product, and the NMR data of compound 2 was reported for the first time.
Annona ; microbiology ; Ascomycota ; chemistry ; genetics ; isolation & purification ; metabolism ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; metabolism ; Endophytes ; chemistry ; genetics ; isolation & purification ; metabolism ; Lactones ; chemistry ; isolation & purification ; metabolism ; Mass Spectrometry ; Molecular Structure

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Male ; Middle Aged ; Musculoskeletal Manipulations ; methods ; Shoulder Dislocation ; therapy ; Young Adult

OBJECTIVETo investigate the best choice of operation opportunity and operation plan for limb fractures combined with severe craniocerebral trauma in children.

METHODSFrom January 2005 to July 2012,36 patients with limb fractures and severe craniocerebral trauma were received,including 24 males and 12 females aged from 1 to 13 years old (mean, 6.1 +/- 3.0). The time from injury to hospital was (18.0 +/- 15.0) h. Glasgow coma score were less than 8 with an average of 6.4 +/- 1.3. AIS-ISS score were 25.9 +/- 8.1. Thirteen patients were open fracture, 23 were closed fracture. Patients were divided into immediate operation group (21 patients) received fracture fixation with 24 h, the average time was (15.0 +/- 7.4) h, and delayed operation group (15 patients) received fracture fixation after 24 h, the average time was (165.4 +/- 114.6) h. All patients were treated by open reduction, and 33 cases by internal fixation, 3 cases were external fixation. Operative time, blood loss, fracture healing time and brain trauma,physical trauma, postoperative rehabilitation situation were observed and evaluated.

RESULTSAll patients were healed at stage I ,and no dead, aggravating of coma, disorders of breathing and circulation occurred during operation. Operative time,blood loss,healing time in immediate operation group was (44.5 +/- 25.3) min, (47.1 +/- 36.5) ml, (2.7 +/- 0.5) months, respectively; while in delayed operation group was (87.0 +/- 40.0) min, (112.7 +/- 67.5) ml, (3.8 +/- 1.2) months,respectively; and there were obvious differences between two groups. There was no siginificant meaning in Glasgow coma score and Fugl-Meyer motor function between immediate operation group (4.7 +/- 0.6, 97.9 +/- 2.7) and delayed operation group (4.7 +/- 0.5, 97.7 +/- 3.9) (t = 0.23, P > 0.05; t = 0.11, P > 0.05).

CONCLUSIONThe condition of limb fractures combined with severe craniocerebral trauma in children is seriously, comfortable surgical opportunity should according to particular case, and immediate operation can performed on the condition of stabled vital signs.

Adolescent ; Child ; Child, Preschool ; Craniocerebral Trauma ; surgery ; Extremities ; injuries ; surgery ; Female ; Follow-Up Studies ; Fracture Fixation ; Fracture Fixation, Internal ; Fractures, Open ; surgery ; Humans ; Infant ; Male

Objective To determine whether morphine having the ability to influence the antiviral effect of lamivudine(3TC)in vitro study.Methods MT2 cells were randomly assigned into morphine+3TC treatment group,morphine+naloxone+3TC treatment group,naloxone+3TC treatment group.Both 3TC and virus control groups were set up.The corresponding MT2 cells were treated with opiates antagonist(naloxone)for 0.5 hours before the 24-hours morphine treatment program was implemented while all of the groups were then infected with equal amounts of cell-free HIV-1 ⅢB strain and 3TC.HIV-1 p24 antigen in culture supernatants collected at days 3,4,5 and 6after infection status was tested and the inhibition of 3TC anti-HIV-1 p24 antigen of various treatment groups calculated.Results Inhibition of 3TC anti-HIV-1 p24 antigen of Morphine+3TC treatment group was the lowest when HIV-1 infected cells at 3rd and 4th day and showed significant difierence (P<0.05)when compared to the 3TC control.However,there was no statistically significant difference among them(P>0.05),when virus was infected the cells at 5th and 6th day.The difference of 3TC anti-HIV-1 p24 antigen inhibition between the morphine+naloxone+3TC treatment group and the naloxone+3TC treatment group was not significant(P>0.05).Similar results were obtained when these two groups were compared to the 3TC control group(P>0.05),respectively.The 3TC anti-HIV-1 p24 antigen inhibition of each treatment group reduced as the time of infection prolonged,showing a significant and time-course effbct.Conclusion The 3TC antiviral effect was reduced by morphine in the early stage of infection,and could be blocked by naloxone.

OBJECTIVETo study the effect of growth factor-primed donor hematopoietic stem cells infusion (GPBSCI) as an early adoptive immunotherapy.

METHODSTwelve patients with high-risk leukemia received prophylactic GPBSCI after allogeneic hematopoietic stem cell transplantation (allo-HSCT). Out of the 12 patients, two were Ph(+) ALL in CR(1), one ALL in CR(2), one refractory ALL, three AML (one in relapse, one refractory disease, one in CR(3)), four CML in advanced stage and one myelodysplastic syndrome-refractory anemia with excess blast (MDS-RAEB).

RESULTSSixteen infusions were performed in the 12 patients, including 5 infusions were performed within +90 days post-SCT. The median mononuclear cells (MNC) and CD3(+) cells infused for GPBSCI before +90 d were 1.00 (0.95 - 1.24) x 10(8)/kg and 0.53 (0.39 - 0.63) x 10(8)/kg, and after +90 d were 2.27 (1.00 - 4.30) x 10(8)/kg and 1.15 (0.55 - 2.10) x 10(8)/kg, respectively. Four patients developed grade I - II acute GVHD, and one grade III acute GVHD. Seven patients developed chronic GVHD, of which four cases were extensive. Two patients had no transfusion related GVHD. No transfusion related pancytopenia was observed. Ten patients survived disease-freely at 563 (415 - 728) days of follow-up. Two patients died of leukemia relapse after GPBSCI.

CONCLUSIONAllo-HSCT with prophylactic GPBSCI could maximize graft-versus-leukemia effect with few fatal complications and might be a potentially curative strategy for hematological malignancy patients with high risk of relapse.

Adolescent ; Adult ; Blood Component Transfusion ; methods ; Child ; Follow-Up Studies ; Hematopoietic Stem Cell Transplantation ; Humans ; Immunotherapy, Adoptive ; Leukemia ; prevention & control ; surgery ; Male ; Middle Aged ; Postoperative Care ; Secondary Prevention ; Tissue Donors ; Transplantation, Homologous ; Treatment Outcome

The aim of study was to investigate the pharmacokinetics and distribution of antithymocyte globulin (ATG) in recipients of partially HLA-matched hematopoietic stem cell transplantation. Fifteen patients with hematological disorders were received hematopoietic stem cell transplantation from partially HLA-matched related donor between October 2003 and October 2004 in the Institute of Hematology and People Hospital, Peking University. All patients including 5 cases of AML, 6 cases of CML, 3 cases of ALL, 1 case of AA were consecutively enrolled in the present study after providing written informed consent. Antithymocyte globulin was administered before allogeneic hematopoietic stem cell transplantation at a dose of 2.5 mg/kg daily for 4 consecutive days (total dose of 10 mg/kg) in the conditioning regimen. The concentration of rabbit ATG in the serum of 15 patients was measured using a new enzyme-linked immunoabsorbent assay (ELISA) for the Fc portion of rabbit IgG. The results showed that the washout phase of ATG elimination was analyzed over 0 - 120 days, results were well-fitted by a single exponential decay giving a mean elimination half-life (t(1/2) beta) of 29.67 +/- 2.60 days. A mean value for the apparent volume of distribution of ATG (V) obtained by analysis of data was 0.12 +/- 0.02 L/kg body weight. The serum concentration of ATG increased up to 44.8% at 5 day before transplantation, peak concentration of ATG was 136.0 +/- 10.3 mg/L, its concentration slowly descend at 0 day, fall up to 7.1 +/- 0.06 microg/ml at 90 day after dosing; t(max) 4.8 +/- 0.7 days; According to AIC (Akaike's information criterion), two compartment model of ATG was estimated. It is concluded that the conditioning regimen containing the dosage of 10 mg/kg of ATG is effective and safely in recipients of partially HLA-matched hematopoietic stem cell transplantation. There is no racial difference in the pharmacokinetics of ATG.
Adolescent ; Adult ; Antilymphocyte Serum ; metabolism ; Child ; Female ; Hematopoietic Stem Cell Transplantation ; Histocompatibility Testing ; Humans ; Male ; Middle Aged ; Tissue Distribution

In the research and development of new drugs, it is very important to investigate the in vitro metabolism of candidate drugs. Traditional models such as liver microsomes have many limitations, while the in vitro model of recombinant human drug metabolizing enzymes is considered as an important and useful approach because of its convenient access, stable activity and low cost. In this study, six major human UDP-glucuronosyltransferases (UGTs) genes (UGT1A1, 1A3, 1A4, 1A6, 1A9 and 2B7) were cloned from human liver cDNA and heterologously expressed in Saccharomyces cerevisiae and baculovirus-infected insect cell. UGT1A1, 1A3, 1A6 and 1A9 were successfully expressed in yeast and showed glucuronidation activity against a variety of different structural types of substrates, but their activities were low. All six UGTs were successfully expressed and exhibited significantly improved glucuronidation activity when Trichopolusia ni cells BTI-TN5B1-4 (High Five) were used as the host. The recombinant human UGTs expressed in insect cells can catalyze the glucuronidation of their specific substrates, and the glucuronidation products were synthesized at milligram-scale with yields of 13%-66% for the first time, of which the structures were identified via MS, ¹H NMR, and ¹³C NMR spectroscopic analysis. Above all, the recombinant human UGTs yeast and insect cell expression systems constructed in this study can be used for in vitro metabolism evaluation in the early stage of new drugs research and development, and also provide a new tool for the synthesis of glucuronide metabolites.

According to published nucleotide sequences, ORF4 gene of barley yellow dwarf virus GAV (BYDV-GAV) was synthesized by reverse transcription-polymerase chain reaction (RT-PCR). The BYDV-GAV ORF4 gene was expressed in baculovirus -insect cell expression system efficiently, and western bolt analysis confirmed its expression product. Confocal laser scanning microscopy showed that GFP: ORF4 fusion protein was associated with the nuclear envelope of insect cells. By expressing the N- and C-terminal regions of ORF4-encoding product (P4) in insect cells combined with structure prediction, it was found that the N-terminal region of P4 containing four a-helices is required for targeting P4 to the nuclear envelope. These results provide a base for biological function of ORF4 gene during systemic infection of BYDV-GAV in host plants further.
Amino Acid Sequence ; Animals ; Baculoviridae ; genetics ; metabolism ; Genes, Plant ; genetics ; Green Fluorescent Proteins ; genetics ; Insecta ; genetics ; metabolism ; Luteovirus ; genetics ; Molecular Sequence Data ; Open Reading Frames ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; metabolism