OBJECTIVE:To establish a method for the contents determination of 4 ingredients in Ligustrum lucidum decoration piece by reference extract method. METHODS:HPLC was performed on the column of XBridge C18 with mobile phase of acetoni-trile-0.1% formic acid(gradient elution) at flow rate of 1.0 ml/min,detection wavelength was 254 nm,column temperature was 30℃and volume injection was 10μl. RESULTS:The linear range was 0.037-0.598 mg for neonuezhenide,0.006-0.101 mg for ac-teoside,0.189-3.023 mg for nuezhenide and 0.314-5.027 mg for specnuezhenide(r≥0.999 0);RSDs of precision,reproducibility and stability tests were no more than 3.8%;recoveries were 98.46%-104.83%(RSD=2.43,n=6),95.55%-104.57%(RSD=3.63, n=6),100.09%-104.39%(RSD=1.45,n=6)and 98.84%-104.97%(RSD=2.02,n=6). CONCLUSIONS:The method is simple, good reproducibility,and can be used for the contents determination of 4 ingredients in L. lucidum decoration piece.

The optimum conditions of baicalin hydrolysis into baicalein by immobilized beta-glucosidase in a two-phase system was studied and the yield was observed. A two-phase system comprising of sodium acetate buffer and chloroform was determined by comparing the solubleness of baicalein in different solvents and partition coefficient of baicalein in related aqueous-organic two-phase system. beta-Glucosidase was immobilized by the crosslinking-embedding method using sodium alginate as the carrier The optimum reaction temperature, pH value, Michaelis constant, the thermal stability and pH stability were assayed. By comparing the yield of baicalin hydrolysis into baicalein by immobilized beta-glucosidase in two-phase system, the optimum reaction conditions were determined-the optimum reaction temperature, pH value and time were 50 degrees C, 5.0 and 10 h, respectively. The yield of baicalein was 85.28%. Compare with one-phase system, two-phase system had an advantage in reaction rate and yield.
Biocatalysis ; Drugs, Chinese Herbal ; chemistry ; Enzyme Stability ; Enzymes, Immobilized ; chemistry ; Flavanones ; chemistry ; Flavonoids ; chemistry ; Hydrolysis ; beta-Glucosidase ; chemistry

Objective To assess the clinical outcomes of glutamine supplementation in neonates.Methods The study was designed as a double-blind randomized controlled trial.Totally 100 infants were randomly divided into 5 groups with 20 infants in each group:control group:standard parenteral nutrition (PN);PNI group:PN supplemented with 0.3 g/ (kg·d) glutamine;enteral nutrition (EN) 1 group:EN supplemented with 0.3 g/ (kg·d) glutamine;PN2 group:PN supplemented with 0.3 g/ (kg·d) glutamine;and EN2 group:EN supplemented with 0.3 g/ (kg·d) glutamine,The primary endpoints were the time to establish full enteral nutrition [defined as receiving 120 ml/ (kg·d) enterally],episodes of gastric residuals,total duration of PN,and mortality.The second endpoints were weight gain and head circumference gain,liver function,renal function,days on ventilator,and length of hospitalization.Results Time to achieve full enteral nutrition,incidence of gastric residuals,duration of PN,weight gain (g/d) ,head circumference gain (cm/w),liver function,renal function,and length of hospitalization were not significantly different among all five groups.Days of mechanical ventilation were significantly decreased in PN1 group and PN2 group when compared with the control group (P ＜ 0.05).Intention-totreat analyses showed that,when compared with the control group,PN1 group:RR:1.053;95% CI:0.952-1.164;ENI group:RR:1.333,95% CI:1.035-1.717;PN2 group:RR:1.053,95% CI:0.952-1.164;EN2 group:RR:1.25,95% CI:1.004-1.556.Conclusions Glutamine supplementation may not be able to decrease the time to achieve full enteral nutrition,the incidence of gastric residuals,and the duration of PN.However,pareneral glutamine supplementation does reduce the days on ventilator in neonates.Also,parenteral glutamine supplementation has no obvious influence on mortality.

Objective: To develop a method for determination of carnitine in serum using HPLC. Methods: The serum proteins were precipitated by acetonitrile and methanol. The organic extract was added derivatizing reagent and the solution was warmed to 60℃ for 2 hours. Atlantis C18 column (4.6 mm?150 mm,5?m)was used as stationary phase and methanol-ispropand-acetonitrile (35∶45∶20,v/v) as mobile phase; flow rate 1.0ml/min, UV wavelengh at 260nm. Results: The typical chromatogram from serum samples showed clear separation of carnitine. Y= 8490X-48200, r= 0.9999. Intra-day RSD was 1.6%-13.2%, and inter-day 1.8%-12.1%. The recovery rate was 98.5%. Conclusion: A method for determination of carnitine in serum using HPLC by adding derivatizing reagent was developed and it is simple, rapid and precise, and can be used for clinical test.

According to the 2010 Chinese pharmacopeia, salt processed and charcoal processed Eucommiae Cortex were pre- pared. HPLC-DAD analysis of the content of the bark and leaf of Eucommiae Cortex showed that the bark of Eucommiae Cortex mainly contained lignans such as pinoresinol glucose and iridoid including genipin, geniposide, geniposidic acid, while the leaf of Eucommiae Cortex consisted of flavonoids such as quercetin and phenolic compound such as chlorogenic acid. The content of pinoresinol diglucoside in the bark of Eucommiae Cortex was about 18 times more than that in the leaf of Eucommiae Cortex. The content of pinoresinol diglucoside in salted and charcoal processed Eucommiae Cortex decreased approximately by 30% and 85%, respectively. The content of genipin, geniposide and geniposidic acid in the bark of Eucommiae Cortex was about 3 times, 23 times, 28 times more than that in the leaf of Eucommiae Cortex. The content of genipin, geniposide and geniposidic acid in salted Eucommiae Cortex were reduced by 25%, 40% and 40%, respectively. The content of genipin, geniposide and geniposidic acid in charcoal processed Eucommiae Cortex were reduced by 98%, 70%, 70%, respectively. The content of caffeic acid in bark of Eucommiae Cortex was about 3 times more than that in the leaf of Eucommiae Cortex. The content of caffeic acid was decreased by about 50% in the salted Eucommiae Cortex. While the content of caffeic acid in charcoal processed Eucommiae Cortex was decreased approximately 75%; the content of chlorogenic acid in bark of Eucommiae Cortex was about 1/6 of that in the leaf of Eucommiae Cortex. The content of chlorogenic acid in salted and charcoal processed Eucommiae Cortex decreased by 40% and 75%, respectively; the content of quercetin in bark of Eucommiae Cortex was only 1/40 of that in the leaf of Eucommiae Cortex. The content of quercetin in salted and charcoal processed Eucommiae Cortex were reduced by 60% and 50%, respectively.
Drugs, Chinese Herbal ; chemistry ; Eucommiaceae ; chemistry ; Flavonoids ; chemistry ; Plant Bark ; chemistry ; Plant Leaves ; chemistry

A method for the determination of S-Adenosyl-L-methionine in fermentation liquid using the high performance liquid chromatography coupled with UV detector (HPLC-UV) was developed.A good linearity was obtained in the range of 0.1～1.0g/L (r = 0.9969) for S-Adenosyl-L-methionine.The average recoveries of S-Adenosyl-L-methionine were 99.89%～101.7% and the relative standard deviations were 0.48%～1.36%.The method is simple and rapid with reproducibility for the determination of S-Adenosyl-L-methionine in ferment liquid.

Arginine Deiminase(ADI) was purified to homogeneity using ammonium sulfate precipitation,Q-Sepharose Fast Flow anion exchange chromatography and SephadexG-75 gel filtration chromatography. This purification protocol resulted in a 34.5-fold purification of ADI with 31.4% final yield. A molecular weight of about 190 kD determined by native gradient polyacrylamide gel electrophoresis. The enzyme has only one kind of 46 kD subunit determined by SDS-PAGE. Combining the results from the two kinds of electrophoresis,the authors deduce that the enzyme may be a tetramer. The optimum pH and temperature for lipolytic activity of ADI was pH 6.5 and 50℃,respectively. It was extremely stable at 45℃ and retained 97.9% of its original activity for 30 min. The stability declined rapidly as soon as the temperature rose over 50℃. ADI was highly stable in the pH range from pH 5-8. ADI acted on L-arginine but not on D-arginine. ADI catabolism was dependent on metal ions. At their adequate concentration,Mn2+,Mg2+ and Co2+ were the effective promoter,while superfluous Zn2+and Co2+ inhibited ADI activity. L-citrulline did not act on ADI,but L-ornithine inhibited ADI activity. The degradation of L-arginine with ADI catalysis was according to simple Michaelis-Menten equation. The Michaelis constant was 3.2686 mmol/L and the maxi-mum velocity was 2.44 ?mol/min.

AIM:To investigate the relationship between the expression of adducin 3 (ADD3) and its splicing isoforms and colorectal cancer (CRC).METHODS:The expression of ADD3, ADD3-Ia and ADD3-Ib in 50 pair of CRC tissues , 20 pairs of colorectal polyp tissues , and 2 CRC cell lines SW480 and SW620 before and after oxaliplatin or fluoroura-cil intervention were detected by real-time PCR.The cell activity was determined by MTT assay , the cell migration ability was evaluated by wound-healing assay , and the cell invasion ability was measured by Transwell assay .RESULTS:The expres-sion levels of ADD3 and ADD3-Ib were decreased in the CRC tissues as compared with the normal mucous (P<0.01), and ADD3-Ia/Ib ratio was increased in the CRC tissues (P<0.01).The expression level of ADD3-Ia was higher in T3-4 group than that in T1-2 group (P<0.05).Reduced expression of ADD3, ADD3-Ia and ADD3-Ib in colorectal polyps was observed compared with the normal tissues (P<0.01).Compared with the SW480 cells, the expression levels of ADD3-Ia and ADD3-Ib were lower (P<0.05) and the ADD3-Ia/Ib ratio was higher (P <0.01) in the SW620 cells.After treated with oxalipla-tin or fluorouracil, the cell activity, migration and invasion in the SW620 and SW480 cells were weakened accompanied by the increases in the expression levels of ADD 3, ADD3-Ia and ADD3-Ib to various certain extents .CONCLUSION:In CRC there is a tendency that ADD3-Ib reduction leads to ADD3 decrease, accompanied by an increased ADD3-Ia/Ib ratio.The expression changes of ADD 3 and its splicing isoforms in the CRC may be relevant to its invasion ability .

Objective To investigate the adjustment methods and evaluation for children after cochlear implantation. Methods 6 children after artificial cochlear implantation were debugged by subjective psychological physical test, objective methods such as electricity induced composite action potential (ECAP), electrically evoked stapedius reflex threshold (ESRT) and electrically evoked auditory brainstem response (EABR). Results All the children got satisfied data. Conclusion For the children after cochlear implantation, objective methods such as ECAP, ESRT, EABR combined with subjective psychological physical test all together are helpful to obtain accurate threshold.

OBJECTIVETo find out the relativity among starch quantity, polysaccharides content and total alkaloid content of Dendrobium loddigesii.

METHODMicroscopy-counting process was applied to starch quantity statistics, sulfuric acid-anthrone colorimetry was used to assay polysaccharides content and bromocresol green colorimetry was used to assay alkaloid content. Pearson product moment correlation analysis, Kendall's rank correlation analysis and Spearman's concordance coefficient analysis were applied to study their relativity.

RESULT AND CONCLUSIONExtremely significant positive correlation was found between starch quantity and polysaccharides content, and significant negative correlation between alkaloid content and starch quantity was discovered, as well was between alkaloid content and polysaccharides content.