A quantitative analysis method of multi-components with a single marker (QAMS) for simultaneous determination of six marker compounds (one from phenolic acids and five from phthalides) in Chuanxiong Rhizoma was established by applying HPLC and using butylidenephthalide as the internal reference substance. And also the feasibility and accuracy of the established method for quality evaluation and application of Chuanxiong Rhizoma were investigated and validated. The analysis was performed with the mobile phase consisting of acetonitrile - 0.2% aqueous formic acid. The flow rate was 1.0 mL . min-1 and the column temperature was maintained at 30 °C. The detection wavelengths were set at 252 nm (for ferulic acid, Z-ligustilide, and butylidenephthalide) and 266 nm (for senkyunolide I, senkyunolide A, and coniferyl ferulate), separately, and 20 µL was injected for analysis with gradient elution. The results showed that there were no significant differences observed between the HPLC-QAMS method and the external standard method (RSD <5%). The relative correction factors were credible (RSD < 5%) in changed chromatographic conditions. The established HPLC-QAMS method can be accurately used for simultaneously evaluating and controlling the quality of Chuanxiong Rhizoma with multi-components.
4-Butyrolactone ; analogs & derivatives ; Acetonitriles ; Benzofurans ; Chromatography, High Pressure Liquid ; Coumaric Acids ; Drugs, Chinese Herbal ; analysis ; standards ; Hydroxybenzoates ; Quality Control ; Rhizome ; chemistry

OBJECTIVETo investigate the effect of water extracts of Coptidis Rhizoma and Evodiae Fructus (CREF) on the proliferation and apoptosis of human gastric carcinoma cells (SGC-7901) and determine the optimal proportion of Coptidis rhizoma to Evodiae fructus.

METHODSThe growth inhibition of SGC-7901 cells treated with the water extracts of CREF of varying proportions was tested with MTT assay. The cell apoptotic rate and mitochondrial membrane potential were analyzed with flow cytometry.

RESULTSThe water extract of CREF with Coptidis Rhizoma: Evodiae Fructus proportions at 1:6, 2:5, 3:4, 4:3, 5:2, and 6:1 all significantly inhibited the growth of SGC-7901 cells after a 24-h or 48-h treatment (P<0.05). The growth inhibition and cell death ratio both exhibited a dose-dependent pattern of Coptidis Rhizoma. Flow cytometry analysis showed that, after treatment of the cells with CREF at the proportions of 1:6, 2:5, 3:4, 4:3, 5:2, and 6:1, the apoptotic rate were (8.50 ∓ 1.59)%, (9.90 ∓ 1.01)%, (17.15∓1.68)%, (21.55 ∓ 1.97)%, (34.10 ∓ 1.06)% and (34.40 ∓ 1.02)%, respectively, all significantly higher than that in the control group [(1.69 ∓ 1.91)%, P<0.05]. JC-1 Kit staining showed that mitochondrial membrane potential of SGC-7901 cells was decreased and the ratio of green to red fluorescence increased significantly after incubation with CREF.

CONCLUSIONCREF can inhibit the growth and induce apoptosis of SGC-7901 cells, and the strongest effect is achieved at the optimal proportion of Coptidis Rhizoma and Evodiae Fructus at 6:1.

Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; drug effects ; Cell Line, Tumor ; Chemistry, Pharmaceutical ; Drug Compounding ; Drugs, Chinese Herbal ; pharmacology ; Evodia ; chemistry ; Humans ; Stomach Neoplasms ; pathology

Objective To develop a multiplex sequence-specific PCR assay for simultaneous screening of 5 types of JAK2 mutations and investigate its clinical application value. Methods Multiplex sequence-specific PCR assay for simultaneous screening of JAK2 V617F, K539L (include 2 types of gene mutations), N542-E543del and E543-D544del mutations were developed. 115 patients with myeloproliferative diseases (MPD) including 61 polycythemia vera (PV) cases, 43 essential thrombocythemia (ET) cases and 11 primary myelofibrosis (MF) cases were analyzed. Results The assay can screen the 5 types of JAK2 mutations efficiently. The detection sensitivity is 1% for JAK2 V617F mutation and 0.1% for the other mutations. JAK2 V617F mutation and JAK2 exon12 mutation were detected in 56 and 3 of the 61 PV samples, respectively. 27 of the 43 ET samples and 6 of the 11 MF samples were JAK2 V617F positive, but no JAK2 exon12 mutation was found in both groups. The 3 cases carrying JAK2 exon12 mutation had the clinical feature of erythrocytosis and erythropoietin-independent erythroid colony formation but without apparent leukocytosis, thrombocytosis and splenectasis. Conclusion The assay can simultaneously screen 5 types of JAK2 mutations with high sensitivity and thus lead to an increased detection rate.

Objective:To investigate the effect of dexmedetomidine on stress response and cellular immune function in patients undergoing radical resection of colon cancer during perioperative period.Methods: 96 cases of colon cancer undergoing radical resection were randomly divided into dexmedetomidine group(group D)and control group(group C).The group D was intubated with in-travenous infusion of dexmedetomidine 0.6 μg/kg,followed by intravenous infusion of 0.3 μg/(kg·h) at the end of the surgery,and the group C was given 0.9% saline at equal volume and rate.The venous blood of patients was collected before anesthesia induction,10 min(T0),immediate postoperative(T1),postoperative 24 h(T2)and postoperative 72 h(T3).Flow cytometry were used to detect T lym-phocyte subsets(CD3+,CD4+,CD8+,CD4+/CD8+)and the percentage of NK cells,and determination of norepinephrine(NE), epinephrine(E),cortisol(Cor),IL-6,IL-10 and TNF-α levels.Records of patients with T0,immediate intubation(Ta),T1,immediate extubation(Tb)of SBP,DBP,MAP,and postoperative adverse reactions were recorded.Results:The SBP,DBP and MAP of group C at Ta,T1,Tbwere significantly higher than that of T0and group D(P<0.05).Compared to those at T0,the levels of CD3+,CD4+,CD8+and CD4+/CD8 at T1and T2were significantly lower in both groups(P<0.05),and the levels of group C at T1,T2,T3were significantly lower than those in group D(P<0.05).The NK cells of group C at T1,T2were significantly lower than that of T0and group D(P<0.05).Compared to those at T0,the levels of IL-6,IL-10 and TNF-α at T1and T2were significantly higher in both groups(P<0.05), and the levels in group C were significantly higher than those in group D(P<0.05).The IL-6 and IL-10 of group C at T3were significantly higher than that of T0and group D(P<0.05).Compared to those at T0,the levels of NE,E and Cor at T1and T2were sig-nificantly higher in both groups(P<0.05),and the levels in group C were significantly higher than those in group D(P<0.05).The NE,E and Cor of group C at T3were significantly higher than that of T0and group D(P<0.05).The proportion of adverse reactions in group D was significantly lower than that in group C(χ2= 4.800,P= 0.028).Conclusion: Dexmedetomidine can inhibit the perioperative stress response and reduce the impact on immune function in patients undergoing radical resection of colon cancer.

FIZZ/RELM is a new gene family named "found in inflammatory zone" (FIZZ) or "resistin-like molecule" (RELM). FIZZ1/RELMα is specifically expressed in lung tissue and associated with pulmonary inflammation. Chronic cigarette smoking up-regulates FIZZ1/RELMα expression in rat lung tissues, the mechanism of which is related to cigarette smoking-induced airway hyperresponsiveness. To investigate the effect of exercise training on chronic cigarette smoking-induced airway hyperresponsiveness and up-regulation of FIZZ1/RELMα, rat chronic cigarette smoking model was established. The rats were treated with regular exercise training and their airway responsiveness was measured. Hematoxylin and eosin (HE) staining, immunohistochemistry and in situ hybridization of lung tissues were performed to detect the expression of FIZZ1/RELMα. Results revealed that proper exercise training decreased airway hyperresponsiveness and pulmonary inflammation in rat chronic cigarette smoking model. Cigarette smoking increased the mRNA and protein levels of FIZZ1/RELMα, which were reversed by the proper exercise. It is concluded that proper exercise training prevents up-regulation of FIZZ1/RELMα induced by cigarette smoking, which may be involved in the mechanism of proper exercise training modulating airway hyperresponsiveness.
Animals ; Lung ; physiopathology ; Male ; Nerve Growth Factor ; metabolism ; Physical Conditioning, Animal ; methods ; Rats ; Rats, Sprague-Dawley ; Respiratory Hypersensitivity ; physiopathology ; Smoking ; physiopathology ; Up-Regulation

Taking the typical metabolic control product-guanosine as an example, the method of metabolic flux shift investigation based on process multi-levels parameter correlation analysis was established. The metabolic pathway, multi-parameter correlation, accumulation of amino acid and organic acid during guanosine fermentation process were integratively analyzed. The metabolic flux shift from HMP to EMP was ascertained, which was assumed to be caused by the accumulation of ammonium ion. The subsequent optimization based on controlling flux distribution between EMP and HMP did improve the yield by 35% when the metabolic flux shift was prevented.
Amino Acids ; metabolism ; Ammonia ; metabolism ; Fermentation ; Guanosine ; metabolism

Metabolic engineering has become a powerful tool for optimization of industrial fermentation processes. Metabolic engineering usually undergoes three steps: construction of a recombinant strain with improved properties, genetic and biochemical analysis of the strain, and identification of target for further improvement. Metabolic fluxes analysis is an important part of the biochemical analysis. Based on the law of mass conservation and assuming pseudo-steady-state for the intermediates in the metabolic pathways, we have quantitatively analyzed the time course of the flux distribution in Bacillus subtilis and used the data to reveal the nature of the so-called "40 hour" phenomenon in fermentation of guanosine, a key raw material for the synthesis of additives for human consumption and animal feeds. The phenomenon refers to the observation that guanosine production, which proceeds at high rate from 12 hour on, declines around 40 hour while consumption of glucose keeps increasing, leading to the lower yield of the nucleoside. Equations based upon the metabolic network of Bacillus subtilis consisted of EMP pathway, HMP pathway, TCA cycle, oxidative phosphorylation pathway and others reactions of the intermediates, was constructed. The equations were solved by using the quantitative data obtained in this study. The air flow and volume, concentration of oxygen and carbon dioxide in the exit-gas were monitored online; the concentration of biomass, glucose and guanosine was analyzed manually; and the concentration of acetate, citric acid, pyruvate, and 17 amino acids were HPLC quantified. The solutions of the equation were proved to be valid, as the experimental data on oxygen consumption agrees with that of predicted form the equation. The results indicated that at 40h of the fermentation process the flux of HMP pathway, which provides the precursor of the nucleoside, decreased while that of EMP pathway and the pathways that generate amino acids and organic acids increased. The shift correlated with the accumulation of NH4+ in the broth. The assimilation of NH4+ is an energy consuming process and could shift the metabolism to the energy generating EMP pathway. Accordingly, measures were taken to prevent the accumulation of NH4+. The interference indeed stopped the metabolism shift and boosted the guanosine production at 30 g/L, 70% higher than the level reported in literature.
Bacillus subtilis ; metabolism ; Fermentation ; physiology ; Guanosine ; metabolism ; Models, Theoretical ; Quaternary Ammonium Compounds ; metabolism

OBJECTIVETo evaluate the effect of glutamine granules on immunofunction in severe burns and trauma patients.

METHODSOne hundred and twenty patients with severe burns, multiple trauma and post operation who met the requirements of the protocol joined this double-blind randomized controlled, multi-center clinical trail. Patients were randomly divided into two groups: placebo control group (P group, 60 patients) and glutamine granules treatment group (GLN group, 60 patients). There was isonitrogenous and isocaloric intake in both groups. GLN and P group patients had been given glutamine granules or placebo (glycine) at 0.5 g.kg(-1).d(-1) for 7 days, respectively. The level of plasma glutamine and some index of immunofunction were determined, and the complication and side effect were also observed.

RESULTSAfter 7 days of taking glutamine granules orally, plasma GLN concentration was significantly higher than that in P group [(593 +/- 185) micromol/L vs (407 +/- 190) micromol/L)] (P < 0.01). IL-2 level, CD(4)/CD(8) ratio, PMN swallow ratio in GLN group were significantly higher than those in P group (P < 0.05-0.01), but the concentration of IgG, IgM, C(3)/C(4) were not significantly different when compared with P group (P > 0.05). In addition, the occurrence of side effect in both groups was seldom.

CONCLUSIONTaking glutamine granules could increase plasma GLN concentration, enhance body immunofunction, and using glutamine granules is safe.

Administration, Oral ; Adolescent ; Adult ; Double-Blind Method ; Female ; Glutamine ; adverse effects ; blood ; therapeutic use ; Humans ; Male ; Middle Aged ; Wounds and Injuries ; blood ; drug therapy ; immunology

OBJECTIVETo investigate the effect of inhibitory and activating KIRs on a cohort of Chinese leukemia patients who received haplo-identical hematopoietic stem cell transplantation (HSCT).

METHODSDonor's inhibitory and activating KIRs and recipient's HLA-C from 47 cases who received haplo-identical HSCT were tested by PCR-SSP. 2 year overall survival (OS), incidence of severe (grade III to IV) acute GVHD (aGVHD) and relapse rate (RR) were analyzed.

RESULTS(1) According to Matched (M) vs Mis-Matched (MM) between donor's inhibitory KIR and recipient's HLA-C1/C2 subgroup, 2 year OS rate in M group [(87.5 +/- 8.3)%] was significantly higher than that in MM group (54.5 +/- 9.0)%, (P = 0.03). Lower incidence of relapse rate was seen in M group than in M/MM groups [0 vs (25.4 +/- 9.5)%, P = 0.05]. In 30 cases of myeloid leukemia patients, there was lower RR in M group than in MM groups [0 vs (35.0 +/- 14.4)%, P = 0.04]. (2) According to the 3 activating KIR genes: KIR2DS1/ KIR2DS2/ KIR2DS3, lower incidence of grade III-IV aGVHD was seen in KIR2DS1 (+) group than in KIR2DS1 (-) group (13% vs 28%, respectively, P > 0.05); and so was done in KIR2DS3 (+) group (11% vs 26%, respectively, P > 0.05). The RR was lower in KIR2DS2 (+) group [0% vs (17.3 +/- 7.1)%, respectively, P > 0.05].

CONCLUSIONSIn our haplo-identical HSCT setting, match between donor's inhibitory KIR and recipient's HLA-C can significantly reduce the incidence of relapse rate and improve OS. Although lower incidences of severe aGVHD are noted in the donors with KIR2DS1 (+) or KIR2DS3 (+), and lower relapse rate is noted in the donors with KIR2DS2 (+) but without statistic difference, no remarkable effects of activating KIRs on OS have been found in our relatively small clinical series.

Adolescent ; Adult ; Child ; Child, Preschool ; Female ; Genotype ; Graft vs Host Disease ; HLA-C Antigens ; genetics ; Hematopoietic Stem Cell Transplantation ; methods ; Humans ; Male ; Middle Aged ; Receptors, KIR ; classification ; Recurrence ; Siblings ; Survival Rate ; Tissue Donors ; Transplantation, Homologous ; Young Adult

OBJECTIVETo evaluate the value of a new platelet function test PFA P2Y (PFA-200) in monitoring clopidogrel treatment for cardiovascular disease in elderly patients.

METHODSFifty-six elderly patients receiving clopidogrel therapy in the Department of Cardiology of General Hospital of PLA from March to August in 2016 and 85 healthy volunteers were recruited for analysis. All the subjects underwent PFA P2Y, LTA (light transmittance aggregometry) and TEG (Thromboelastograph) tests, and Spearman correlation coefficients were used to test the associations between test results. The agreement among the 3 platelet function test methods was assessed using Cohen's kappa coefficient.

RESULTSCorrelation coefficient (r) was -0.701 (P<0.001) between PFA P2Y and LTA, and 0.475 (P<0.001) between PFA P2Y and TEG. The agreement was 75% between PFA P2Y and LTA and 67.9% between PFA P2Y and TEG. The κ value was 0.434 (P=0.001) between PFA P2Y and LTA and 0.242 (P=0.046) between PFA P2Y and TEG. With ADP-induced maximum platelet aggregation rate of LTA >50% as the laboratory clopidogrel resistance, the cut-off value of PFA P2Y was 119 s (AUC=0.733) with a sensitivity of 75.6% and a specificity of 73.3%.

CONCLUSIONPFA P2Y has a moderate correlation and agreement with LTA, but has a poor correlation and agreement with TEG. PFA P2Y can be useful for assessing the effects of clopidogrel therapy and the association of the cut-off value (119 s) with the long-term clinical ischemic events needs be confirmed in further study.

Biological Assay ; Blood Coagulation Tests ; Blood Platelets ; Cardiovascular Diseases ; drug therapy ; Humans ; Platelet Aggregation ; Platelet Aggregation Inhibitors ; therapeutic use ; Platelet Function Tests ; Sensitivity and Specificity ; Ticlopidine ; analogs & derivatives ; therapeutic use