OBJECTIVETo analyze characterization and determination of nitrogen in the preparation of Qingkailing injection and its intermediate products.

METHODHitich amino acid auto analyzer was used, with the packed analysis column (2.6 mm x 50 mm) and the type of was Hitich 2622 SC resin. The speed of buffer solution and ninhydrin colorimetric solution were 0.4 mL x min(-1) and 0. 3 mL x min(-1) respectively. Program heating was used for controlling column temperature, from 57 degrees C (0.0 min) to 65 degrees C (36 min) to 57 (50 min). The reaction temperature was set at 130 degrees C.

RESULTFree and binding amino acid the existenceare the main form of nitrogen is amino acid in Qingkailing injection and its intermediate products. The total contents of amino acid in the preparation of Qingkailing injection and its intermediate products, including hydrolyzed solution which is made from neutralization of Concha Margaritifera solution extracted by diluted sulfuric acid and Cornu Bubali solution extracted by diluted sodium hydroxide, aqueous solution of Radix Isatidis extract, 4-blended solution, 6-blended solution and 8-blenede solution, were 59.56%, 24.88%, 41.84%, 13.49, 14.63% respectively. The type of bonded amino acid was founded in the preparation of Qingkailing injection and its intermediate products, including hydrolyzed aqueous solution of Radix Isatidis extract, 4-blended solution, 6-blended solution and 8-blenede solution, and the contents were 9.33%, 15.07%, 16.85%, 19.94% and 19.55%, respectively.

CONCLUSIONThe main resource of the total nitrogen was Bubalus bubalis L. and Isatis indigotica Fort.

Amino Acids ; analysis ; chemistry ; Animals ; Buffaloes ; metabolism ; Chromatography ; methods ; Drugs, Chinese Herbal ; chemistry ; Isatis ; chemistry ; Nitrogen ; analysis ; chemistry

We investigated the disposition kinetics and urinary excretion of cefpirome in buffalo calves after a single intravenous administration of 10 mg/kg. Also, an appropriate dosage regimen was calculated. At 1 min after injection, the concentration of cefpirome in the plasma was 57.4 +/- 0.72 microgram/ml, which declined to 0.22 +/- 0.01 microgram/ml at 24 h. The cefpirome was rapidly distributed from the blood to the tissue compartment as shown by the high distribution coefficient values (8.67 +/- 0.46/h), and by the drug's rate of transfer constant from the central to the peripheral compartment, K12 (4.94 +/- 0.31/h). The elimination halflife and the volume of distribution were 2.14 +/- 0.02 h and 0.42 +/- 0.005 l/kg, respectively. Once the distribution equilibrium was reached between the tissues and plasma, the total body clearance (ClB) and the ratio of the drug present in the peripheral to the central compartment (T/P ratio) were 0.14 +/- 0.002 l/kg/h and 1.73 +/- 0.06, respectively. Based on the pharmacokinetic parameters we obtained, an appropriate intravenous cefpirome dosage regimen for treating cefpiromesensitive bacteria in buffalo calves would be 8.0 mg/kg repeated at 12 h intervals for 5 days, or until persistence of the bacterial infection occurred.
Animals ; Buffaloes/*metabolism/urine ; Cephalosporins/administration & dosage/*pharmacokinetics/*urine ; Injections, Intravenous/veterinary ; Kinetics ; Metabolic Clearance Rate/physiology

The present study was planned to investigate the pharmacokinetics of ceftriaxone in experimentally induced febrile buffalo calves (n = 5). The fever was induced by intravenous injection of E.coli lipopolysaccaride (1 microgram/kg). To study the pharmacokinetics, ceftriaxone was administered at the dose rate of 10 mg/kg body wt. in all animals. At 1 min, the peak concentration of ceftriaxone was 79.4 +/- 2.37 microgram/ml and the drug was detected up to 6 h. The elimination rate constant was 0.35 +/- 0.02 /h and elimination half-life was 2.04 +/- 0.14 h. The apparent volume of distribution (Vd(area)) and total body clearance (ClB) were 1.21 +/- 0.15 l/kg and 0.41 +/- 0.03 l/kg/h, respectively. To maintain a minimum therapeutic concentration of 1 microgram/kg, a satisfactory dosage regimen of cefriaxone in febrile buffalo calves is 19 mg/kg followed by 18 mg/kg at 8 h intervals.
Animals ; Anti-Bacterial Agents/administration&dosage/*pharmacokinetics ; Area Under Curve ; Buffaloes/*metabolism ; Ceftriaxone/administration&dosage/*pharmacokinetics ; Drug Administration Schedule/veterinary ; Fever/drug therapy/*metabolism/*veterinary ; Half-Life ; Lipopolysaccharides/administration&dosage ; Male ; Metabolic Clearance Rate

Metagenomic cosmid libraries containing 1.26 x 10(5) clones, covering about 4.8 x 10(6) kb metagenomic DNA of uncultured microorganisms from the contents of buffalo rumens were constructed, and 118 independent clones expressing beta-glucosidase activity were isolated from the libraries. Screening of these clones showed that eight clones expressed relatively higher beta-glucosidase activity at pH 5.0 and 37 degrees C. One out of the eight clones was subcloned. Sequencing analysis showed that an open reading frame (ORF) of 2223 bp, termed umcel3G, potentially encodes a beta-glucosidase. The encoded product shared highest homology with a beta-glucosidase from Bacillus sp. at 60% identity and 73% similarity. The umcel3G was over-expressed in Escherichia coli and the size of the translated product Umcel3G on SDS-PAGE was in agreement with the predicted molecular mass. Zymogram analysis showed that Umcel3G exhibited beta-glucosidase activity, confirming that this ORF encodes a beta-glucosidase. The Umcel3G, purified with Ni-NTA column, exhibited optimal activity at pH 6.0-6.5 and 45 degrees C. Certain ions such as Ca2+, Zn2+ had significant positive effect on the activity of Umcel3G. However, some ions such as Fe3+, Cu2+ gave significant inhibitory effect on the enzyme. The Ni-NTA purified recombinant beta-glucosidase Umcel3G had a specific activity of 22.8 IU/mg at pH4.5, 35 degrees C and at the presence of 5 mmol/L Ca2+, indicating that this enzyme has potential applications in the fermentative production of ethanol by simultaneous saccharification and cofermentation (SSCF) of lignocelluloses.
Animals ; Bacteria ; enzymology ; genetics ; Buffaloes ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Open Reading Frames ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; isolation & purification ; metabolism ; Rumen ; microbiology ; beta-Glucosidase ; biosynthesis ; genetics ; isolation & purification

Effect of sodium nitroprusside (SNP), a nitric oxide (NO) donor, on in vitro survival, growth, steroidogenesis, and apoptosis of buffalo preantral follicles (PFs) was investigated. PFs (200~250 microm) were isolated by micro-dissection and cultured in 0 (control), 10(-3), 10(-5), 10(-7), and 10(-9) M SNP. To examine the reversible effect of SNP, PFs were cultured with 10(-5) M SNP + 1 mM Nomega-nitro-L-arginine methyl ester (L-NAME) or 1.0 microg hemoglobin (Hb). The results showed that greater concentrations of SNP (10(-3), 10(-5), 10(-7) M) inhibited (p < 0.05) FSH-induced survival, growth, antrum formation, estradiol production, and oocyte apoptosis in a dose-dependent manner. However, a lower dose of SNP (10(-9) M) significantly stimulated (p < 0.05) the survival, growth, antrum formation, follicular oocyte maturation, and stimulated progesterone secretion compared to the control. A combination of SNP + L-NAME promoted the inhibitor effect of SNP while a SNP + Hb combination reversed this effect. Nitrate and nitrite concentrations in the culture medium increased (p < 0.05) in a dose-dependent manner according to SNP concentration in the culture medium. At higher concentrations, SNP had a cytotoxic effect leading to follicular oocyte apoptosis whereas lower concentrations have stimulatory effects. In conclusion, NO exerts a dual effect on its development of buffalo PFs depending on the concentration in the culture medium.
Animals ; *Apoptosis ; Buffaloes/*physiology ; Estradiol/biosynthesis ; Female ; Follicle Stimulating Hormone/metabolism ; NG-Nitroarginine Methyl Ester/pharmacology ; Nitrates/pharmacology ; Nitric Oxide/*metabolism ; Nitric Oxide Donors/pharmacology ; Nitrites/pharmacology ; Nitroprusside/pharmacology ; Oocytes/cytology/drug effects/growth & development/metabolism ; Ovarian Follicle/*cytology/drug effects/growth & development/*metabolism ; Progesterone/biosynthesis