Cofactor engineering, a vital part of metabolism engineering, changes the redox cofactor regeneration approach. Its main goal is to rebuild the components of metabolic products. The bioconversion of xylose for the production of ethanol is being studied intensively because ethanol is an alternative energy source and a potential liquid fuel. Saccharomyces cerevisiae has been traditionally used in producing ethanol from fermentable sugars but it cannot utilize xylose, only its isomer xylulose. Introduction of the xylose fermentation pathway from Pichia stipitis into S. cerevisiae enables xylose utilization in recombinant S. cerevisiae, but the ethanol yields of xylose fermentation with recombinant S. cerevisiae has been low and large amounts of the byproduct xylitol are produced. The major reason is that the catabolism of xylose with the fungal pathway leads an imbalance of redox cofactor. The process of the catabolism of xylose requires NADPH and NAD~+, both of which have to be regenerated in separated processes. More and more attention has therefore focused on the redox cofactor balance in S. cerevisia. The research progress of cofactor engineering to solve the imbalance of redox cofactor in xylose metabolism recombinant S. cerevisiae was introduced. This included expression of transhydrogenase, increasing the utilization of NADPH, and achieving the anaerobic reoxidation of NADH. Reversing the cofactor specificity of enzymes is another effective way.

This research is to establish an HPLC method for the simultaneous determination of ophiopogonin D, ophiopogonin D', ophiopogonin C, deacetylophiopojaponin A and ophiogenin-3-O-α-L-rhamnosyl-(1-->2)-β-D-glucoside in Ophiopogonis Radix. HPLC-ELSD analysis was performed on a Kromasil 100-5 C₁₈ column (4.6 mm x 250 mm, 5 µm), with the mobile phase of acetonitrile (A) -water (B) in gradient elution mode (0-45 min, 35%-55% A), at a flow rate of 1 mL · min⁻¹. The column temperature was 35 °C and the drift tube temperature was 100 °C in a gas flow rate of 3.0 L · min⁻¹. The result showed that baseline of all the 5 constituents was well separated, and every constituent had wide linearity range and good linear relation (r > 0.999). The recovery rate was between 95.75% and 103.1%. The new established method for simultaneous determination of saponin constituents in Ophiopogonis Radix was sensitive and has good, repeatability. It could be applied to quality evaluation of Ophiopogonis Radix.
Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Ophiopogon ; chemistry ; Plant Roots ; chemistry ; Saponins ; chemistry ; isolation & purification

Objective:To evaluate the clinical value of Truview~(TM)EVO_2 optic laryngoscope by comparing it with the Macintosh laryngoscope in patients receiving cervical vertebral surgery.Methods:One hundred patients scheduled for elective cervical vertebral surgery were enrolled in this randomized crossover study.After induction,the patients'glottis in group A (n=50)was displayed by Macintosh laryngoscope and the Cormack-Lehane(C/L)grade was recorded,and then optic laryngoscope was employed to display the laryngeal structure.The order of laryngoscopy attempts was reversed in group B(n= 50).Parameters recorded included demographics,airway assessment feat ures(BMI,thyromental distance,mandibular size,mouth opening,mallampati oropharyngeal scale,and neck movement),C/L grade,laryngoscopic force applied,duration of intubation, difficulties of laryngeal view and injury of upper airway.Results:There were no significant difference in demographics,airway assessment features,C/L grade and duration of intubation between the 2 groups,whereas the laryngoscopic force in group A was significantly lower than that in group B(P

Objective:To examine the feasibility of using cerebral state index(CSI)for monitoring the sedation depth during target-controlled infusion(TCI)with propofol and remifentanil.Methods:Forty-four consenting ASAⅠorⅡpatients(aged 18-60 years)undergoing elective surgery under general anesthesia were randomly divided into 4 groups(n=11 each)according to the target effect-site concentrations of remifentanil administered by TCI during induction of anesthesia.The target effect-site concentrations of remifentanil of R_0,R_2,R_4,and R_6 groups were 0,2 ng?ml~(-1),4 ng?ml~(-1),and 6 ng?ml~(-1),respectively. Anesthesia was induced by TCI with remifentanil and propofol.CSI and bispectral index(BIS)were used to measure the sedation depth.The initial effect-site propofol concentration(PCe)was 1.5?g?ml~(-1),which was increased by 0.5?g?ml~(-1) every 4 min.The modified OAA/S score(5=alert,1=does not respond to prodding),loss of eyelash reflex(LOR eyelash)and loss of response to electric tetanie stimulation(LOR tetanic)were compared against CSI,BIS and PCe(calculated effect-site propofol concentration).Correlation coefficients were calculated between CSI and other parameters.Results:The 4 groups were comparable with respect to the ages and bodyweights.CSI and BIS values were higher but PCe value were lower at LOR eyelash and LOR tetanic in R_2,R_4,and R_6 than those in the R_0 group(P

To investigate the effect of Tanreqing injection on immune activity of peripheral blood lymphocytes of patients with lung cancer. The peripheral blood lymphocytes of patients with lung cancer and healthy persons were separated by the density gradient centrifugation method for subsequent experiments, with those from healthy persons as the positive control. The effect of Tanreqing injection on stimulating the proliferation of lymphocytes with phytohemagglutinin (PHA) was determined by MTT method. The effect of Tanreqing injection on the lymphocyte secretions of IFN-γ and TNF-α and the subset ratio of lymphocytes cultured separately or with Tanreqing injection of different concentrations were examined by ELISA and flow cytometry (FCM) respectively. In addition, the LDH release assay was used to detect the cytotoxicity of cytotoxic T cells (CTL) and natural killer cells (NK). According to the findings, all of immunological indexes of lymphocytes from patients with lung cancer were weaker than that of healthy persons, but with the obvious increases in proliferation activity and IFN-γ and TNF-α secretions of lymphocytes co-cultured with Tanreqing Injection (P < 0.05). Among lymphocyte subsets co-cultured with Tanreqing Injection, CD3+, CD3+ CD4+ and CD3- CD16 + 56+ cell ratios notably increased, whereas CD4+ CD25+ Treg cell ratio obviously decreased (P < 0.05). In the meantime, Tanreqing injection can markedly promote the cytotoxicities of CTL and NK (P < 0.05). In conclusion, Tanreqing injection shows a significant effect in promoting the immune activity of lymphocytes from patients with lung cancer and their anti-tumor immunity.
Cell Proliferation ; drug effects ; Cells, Cultured ; Drugs, Chinese Herbal ; administration & dosage ; Humans ; Interferon-gamma ; genetics ; immunology ; Killer Cells, Natural ; drug effects ; immunology ; Lung Neoplasms ; drug therapy ; genetics ; immunology ; physiopathology ; T-Lymphocytes, Cytotoxic ; drug effects ; immunology ; Tumor Necrosis Factor-alpha ; genetics ; immunology

OBJECTIVETo observe the impact of bone morphogenetic protein-2 (rhBMP-2) on bone marrow stromal cells (BMSCs) osteogenesis in vitro and vascular endothelial growth factors (VEGF) expression in bone osteoporotic to prevent and treat the osteoporosis.

METHODSTwenty 6-month-old female SD rats weighted (300±20) g underwent bilateral ovariectomized. At 3 months after operation, dual-energy X-ray absorptiometry was used to measure bone mineral density of rats,the values were compared with preoperative to ensure the model successfully, and the osteoporosis rats' BMSCs were cultured by bone marrow adherent cultured and the BMSCs morphology was observed under a phase contrast microscope upside down. The osteoporosis rats' BMSCs at the 2nd generation (p2) were randomly divided into experimental and control groups and were added complete medium (containing rhBMP-2) and osteogenic induced liquid, respectively. Two weeks later, the formation of cell calcium nodules were detected by Alizarin red staining method,alkaline phosphatase activity was measured by enzyme standard instrument and the expression of VEGF was detected by RT-PCT method.

RESULTS(1)Whole body bone mineral density of rats before and after operation were (0.179±0.007), (0.158±0.006) g/cm2,there was statistically significant (t=4.180,P< 0.05). (2)Alizarin red staining at 2 weeks after osteogenesis induced by BMSCs (P2) in the experimental group had more strong dyeing effect than the control group obviously. (3)Alkaline phosphatase activity at 2 weeks after osteogenesis induced by BMSCs (P2) of the experimental group (15.62±1.27) ug/gprot was significantly higher than that of the control group (8.62±0.93) ug/gprot,there was statistically significant (t=7.709, P<0.01). (4)The expression of VEGF at 2 weeks after osteogenesis induced by BMSCs (P2) of the experimental group 3.723±0.143 was significantly higher than that of the control group 0.950±0.072, there was statistically significant (t=29.462, P<0.01).

CONCLUSIONRhBMP-2 can improve the in-vitro osteogenesis ability of ovary osteoporosis rat BMSCs, promote the VEGF expression of osteogenesis factor. Regulating the VEGF expression may be one of the mechanisms of BMP-2 to participate in bone metabolism.

Animals ; Bone Density ; Bone Morphogenetic Protein 2 ; genetics ; metabolism ; Cells, Cultured ; Female ; Humans ; In Vitro Techniques ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Osteogenesis ; Osteoporosis, Postmenopausal ; genetics ; metabolism ; physiopathology ; Rats ; Rats, Sprague-Dawley ; Vascular Endothelial Growth Factor A ; genetics ; metabolism

Decompression, Surgical ; methods ; Humans ; Male ; Microsurgery ; Middle Aged ; Vertigo ; surgery ; Vestibulocochlear Nerve ; surgery

Objective To investigate the hemodynamics of increasing portal venous pressure(PVP) in hepatocellular carcinoma patients complicated with hepatic arterioportal vein fistulas (HAPVF)and the diagnosis and therapy of intractable upper gastrointestinal hemorrhage caused by HAPVF.Methods One hundred and fifteen cases of hepatocellular carcinoma with upper gastrointestinal hemorrhage were checked by hepatic arteriography and were treated through orifices embolization in cases with severe HAPCF by coils and/or ethanol. Results Twenty-six out of 31 patients suffering intractable upper gastrointestinal hemorrhage have severe HAPVF(the main stem of portal veins are visible).However,there are only 15 patients with light HAPVF among the 84 patients who have mild upper gastrointestinal hemorrhage (the main stem of portal veins are invisible).After the embolization,all of the 26 patients who have severe HAPVF stopped bleeding.Among them,the main stem of hepatic arteries are occluded in 2 patients. Conclusion The existence of severe HAPVF should be taken into consideration when intractable upper gastrointestinal hemorrhage occurs in hepatocellular carcinoma patients,and it can be diagnosed through hepatic artery DSA.Orifice embolization is the most effective method for such kind of hemorrhage.

Objective To investigate the effects of different degrees of hemodilution with the artificial plasma substitutes most commonly used in clinical practice on coagulation in vitro. Methods Ten male ASA physical status I volunteers aged (28.8?1.6) yr and weighing (66?8) kg were enrolled. Venous blood samples obtained from each volunteer were diluted with 10% HES (200 000/0.4-0.55), 4 % succinylated gelatin (gelofusine GEL), 3.5 % polygeline (Haemaccel HAE) and lactated Ringer's solution (RL) by 33% [blood: diluent(B:D) =2:1], 50% (B:D=1:1) and 66% (B:D=1:2). Coagulation of the undiluted blood (control) and diluted blood was measured with sonoclot coagulation and platelet function analyzer (SCT) and by routine coagulation tests. The parameters measured included activated clotting time (ACT), clot rate, platelet function (PF), Hct, platelet count (Pit), plasma fibrinogen concentration (Fig) and activated partial thromboplastin time (APTT) . Results (1) At 33 % dilution ACT was significantly shortened in all the four groups as compared with control value; at 50 % dilution ACT was significantly shorter than the control in RL, HES and HAE groups; at 66 % dilution ACT was significantly prolonged in HES group. (2) Clot rate was significantly decreased at 33 % dilution only in HES group but was significantly decreased in all the four groups at 50 % and 66 % dilution. (3) PF decreased significantly only in GEL group at 33 % dilution but was significantly decreased in GEL and HES groups at 50 % and 66 % dilution. (4) With increasing dilution Hct, Pit and Fig gradually decreased and APTT was prolonged. Conclusion Coagulation changes are closely related to the degrees of dilution. At 33 % dilution, the three artificial plasma substitutes tested do not significantly affect hemostasis. At 50 % and 66 % dilution coagulation is badly impaired, but there are differences among the substitutes used for dilution. HAE impairs oagulation least as it contains higher calcium

Objective To analyze the relationship between the radiation doses to mediastinal lymph nodes regions and the regional failure patterns when involved field radiation therapy (IFRT) was used for limited-stage small cell lung cancer (SCLC).Methods The mediastinal lymph node regions (group 1 to 10) of the iimited-stage SCLC patients received definitive radiotherapy were contoured in treatment planning system.The intentional or incidental radiation doses to each lymph node regions were recorded.In-field recurrence,marginal recurrence and out-of-field recurrence were respectively defined as the volume of failed lymph nodes located within the 80％ iso-dose lines,in the 80％-20％ iso-dose lines and beyond the 20％ iso-dose lines of prescribed doses.Results A total of 1 216 lymph node regions in 76 patients were contoured.The median follow-up time was 17.4 months.At diagnosis,lymph node regions with metastatic rates ＞50％ were 4R (68.7％),4L (57.9％),10R (57.9％),2R (56.6％) and 7 (51.3％).The positive lymph node regions were all subjected to prescribed doses.The lymph node regions that received incidental radiation doses of more than 3 000 cGy were:3P,4L,7,6,4R,5,2L.The median lymph node failure-free time was 9.8 months.In this study,only 1 patient developed out-of-field mediastinal lymph nodes failure.The rest of out-of-field recurrences and marginal recurrences were developed in the supraclavicular regions or contralateral hila.Conclusions When IFRT is used to treat mediastinal lymph node regions for patients with SCLC,negative mediastinal regions can be subjected to considerable incidental radiation doses.Out-of-field recurrences of the mediastinal lymph node regions are rare.This is contributed by the incidental radiation dose to these regions.