To develop a LC-MS/MS method for the determination of protocatechuic acid, protocatechuic aldehyde, salvianolic acid A, salvianolic acid B, cryptotanshinone and tanshinone II(A) in rat plasma and brain. The plasma and brain samples were precipitated with ethyl acetate, then were separated on an Agilent eclipse plus-C18 column (2.1 mm x 50 mm, 3.5 microm) using acetonitrile (consisting of 0.1% formic acid) and water (consisting of 0.1% formic acid) as mobile phase in gradient elution mode. The mass spectrometer was operated under both positive and negative ion mode with the ESI source, and the detection was performed by MRM. The transition of 154.3/153.1 m/z for protocatechuic acid, 137.3/108 m/z for protocatechuic aldehyde, 493.0/295.2 m/z for Salvianolic acid A, 718.0/520.0 m/z for salvianolic acid B, 321.4/152.3 m/z for chloramphenicol, 297.4/254.3 m/z for cryptotanshinone, 295.5/249.3 m/z for tanshinone II(A) and 285.2/154.0 m/z for Diazepam. The calibration curves in the range of 0.625-1 000 microg x L(-1) for protocatechuic acid and protocatechuic aldehyde, 1.25-1 000 microg x L(-1) for salvianolic acid A, 2.5-1 000 microg x L(-1) for salvianolic acid B, 0.15-1 000 microg x L(-1) for cryptotanshinone, 0.625-1 000 microg x L(-1) for tanshinone II(A) are with good linearityin rat plasma and brain. The analysis method is sensitive, simple, and suitable enough to be applied in the pharmacokinetic study of the 6 main components. Animal testing gives the lgBB of the drugs and further studies of the 6 components cross the blood-brain barrier can be carried out.
Animals ; Benzaldehydes ; administration & dosage ; blood ; pharmacokinetics ; Benzofurans ; administration & dosage ; blood ; pharmacokinetics ; Blood-Brain Barrier ; metabolism ; Brain ; metabolism ; Caffeic Acids ; administration & dosage ; blood ; pharmacokinetics ; Catechols ; administration & dosage ; blood ; pharmacokinetics ; Chromatography, Liquid ; methods ; Diterpenes, Abietane ; administration & dosage ; blood ; pharmacokinetics ; Hydroxybenzoates ; administration & dosage ; blood ; pharmacokinetics ; Injections, Intravenous ; Lactates ; administration & dosage ; blood ; pharmacokinetics ; Phenanthrenes ; administration & dosage ; blood ; pharmacokinetics ; Plant Preparations ; administration & dosage ; blood ; pharmacokinetics ; Rats ; Reproducibility of Results ; Salvia miltiorrhiza ; chemistry ; Tandem Mass Spectrometry ; methods

Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Cisplatin ; administration & dosage ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Fluorouracil ; administration & dosage ; Humans ; Intestinal Neoplasms ; drug therapy ; Lung Neoplasms ; drug therapy ; Male ; Phytotherapy ; Tetrahydrofolates ; administration & dosage

Objective To study the normal angiographic anatomy and variations of rabbit hepatic vessels,and explore the optimal method for hepatic artery catheterization.Methods 30 rabbits were divided into two groups randomly.Modified surgical method and interventional method were used to catheterize hepatic artery respectively,and followed by angiography to demonstrate the normal anatomy and variations of rabbit celiac artery,hepatic artery and portal vein.Results The route and distribution of rabbit celiac artery and hepatic artery were very different from human's.The commonly seen variation showed the differences in branching bifurcation of hepatic-gastric artery,with the incidence of 13.3%.The rates of successfully hepatic artery catheterization with surgical and interventional methods were 86.6%(13/15)and 80%(12/15)respectively (P＞0.05).The surgical method will not be successful,whenever there's variation.Conclusion The normal anatomy and variation of rabbit celiac artery and hepatic artery are quite different from human's.Both surgical and interventional catheterizations could be rather successful but possessing advantages and disadvantages of each its own.

Objective To investigate the changes and pathogenic significance of serum interleukin-12p70 (IL-12), intefferon-γ,(IFN-γ) and IL-4 in the course of hemorrhagic fever with renal syndrome(HFRS). Methods Twenty five eases were divided into mild group (14 eases) and severe group (11 cases) according to the severity of illness. Blood samples were collected in various stages(fever, hypotensian and oliguria,diuresis stage). Serum IL-12 and IFN-γ levels were determined by enzyme-linked immunoserbent assay(ELISA), IL-4 by radioimmunoassay (RIA), blood urea nitrogen (BUN) and platelet by automatic biochemical analyzer and blood analyzer. Results Serum IL-12 levels in mild and severe groups were significantly different during various stages of HFRS (F=5.765, P<0.01). The IL-12 level of both patient groups significantly increased(P<0.01) in fever[ (0.87±0.38), (1.08± 0.77)μg/L], hypotension and oliguria [ (0.77±0.21), (2.11±2.13)μg/L] ,and diuresis stage [ (1.42±1.10), (1.20±0.88)μg/L], compared with control group [(0.56±0.10)μg/L]. In various stages, IFN-γ levels of both case groups were respectively (8.04±13.05), (5.94±8.24), (15.95±18.05), (4.41±4.10), (1.09±1.24), (1.38±1.74), (1.12±1.26), (0.19±1.29)μg/L, and the difference was statistically significant compared with control [ (0.27±0.15)rig/L]. K,-4 levels did not change significantly in the stages(F=0.682, P0.05), while the ratios of IFN-γ and IL-4 contents in mild and severe cases were significantly higher than control [(0.36±0.26) μg/L] in fever[ (2.46±3.52), (16.92±22.77)p.g/L], hypotension and oliguria[(2.52±2.72), (1.77±2.06) μg/L],diuresis stage [(1.45±2.28), (2.32±3.98)μg/L], the difference had statically significant (P<0.05 or 0.01).The curve of IL-12 was similar to that of BUN, but was contrary to blood platelet count. Conclusions The elevated levels of IL-12 and IFN-γ, with the imbalance of Th1/Th2 might be the main cause of systemic inflammatoryresponse and involved in the pathogenesis of HFRS.

Child, Preschool ; Humans ; Male ; Neurofibromatosis 1 ; genetics ; Pedigree

The deep fermentation technique of Tricholoma matsutake is systemically studied in this paper firstly. The best culture determined by orthogonal test is 3g/L of cornflour, 1g/L of glucose, 1g/L of bean cake flour, 1mL/L of corn steep liquid, 1g/L of KH 2PO 4. The best fermenting condition is: 25℃, rotating speed 160 r/min, pH5.0,inoculating amount 10%, 120mL culture medium per 500mL flask. Under these conditions, the mycelia reach 12.94g/L after fermenting 12d.

Producing cellulase conditions such as different temperature,inoculum size,liquid level and pH level by Trichoderma aureoviride in the shaking bottle were studied by orthogonal design method. The results showed that the main factor affecting the producing cellulase was temperature among the orthogonal conditions.The optimum conditions were as follows:cultivating solution initial pH was 6, cultivating temperature was 28℃,inoculation size was 8%,liquid level was 40 mL in 150 mL triangle bottle,rotation speed was 170 r/min.

Objective:To purify HLA-DR molecules. Methods: Anti-HLA-DR antibody L243 was purified and coupled with CNBr activated Sepharose 4B gel. Immunoaffinity column was used to purify HLA-DR molecules. Results:Twenty micrograms of HLA-DR molecules were isolated from about 5 g Epstein-Barr virus-transformed human B lymphoblastoid cell line RAJI lysates by affinity chromatography. The purified HLA-DR molecules existed in α/β heterodimers form and could bind to conformation-dependent antibody L243. These HLA-DR molecules were separated into two strands,α and β,by boiling denaturation. These results are the basis for studying MHC Ⅱ binding peptide motif and CD4＋ T cell epitopes of antigens in future.

Objective To evaluate the role of early cleavage embryo in combination with embryo growth rate and morphology scoring in embryo selection in in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) cycles. Methods Six hundred and ten IVF/ICSI cycles were randomly assigned to group A(269 cycles) and group B(341 cycles). In group A, transferred embryos were chosen according to embryo growth rate and morphology scoring by 72 h(D3) after fertilization, while early cleavage embryo was added to the selecting system in group B. The pregnancy rate and implantation rate were compared between two groups, and the clinic outcomes were compared between transfers with early cleavage embryos and without early cleavage embryos in group B. Results The pregnancy rate and implantation rate in group B were significantly higher than those in group A (P < 0.05). Transfers with early cleavage embryos also achieved much higher pregnancy rate and implantation rate in group B (P < 0.01). Conclusion Compared with embryo growth rate and morphology scoring, early cleavage embryo in combination with embryo growth rate and morphology scoring can improve the clinical outcomes in IVF/ICSI cycles.

OBJECTIVETo investigate the efficacy mechanisme of chicory extract interventing abdominal obesity rat from the aspect of gut bacteria.

METHODMale SD rats were randomly divided into five groups, namely the normal group, model group, large and small dose group of chicory and the fenofibrate group. Normal group was given deionized water, the other group was given fructose water and give the medical treatment of chicory and fenofibrate. Assay triglycerides, total cholesterol, LDL and HDL by biochemical methods and measure body weight and abdominal circumference and microscopicly observe the count changes of gut bacteria through real-time PCR method.

RESULTCompared with normal group, the triglyceride level and abdominal circumference were significantly higher (P < 0.05), weight and high-density lipoprotein increased but no significant changes and E. coli, lactobacillus increased significantly. Compared with model group, chicory extract large and small dose group and the fenofibrate group can significantly reduce triglyceride levels (P < 0.05), reduce the number of E. coli and Lactobacillus and increase the number of bifidobacteria. The fenofibrate group can significantly reduce total cholesterol and high-density lipoprotein levels.

CONCLUSIONThe chicory's treatment effect on abdominal obesity is significant. The efficacy mechanisme intervention abdominal obesity may be related to the reduction of the number of lactic acid bacteria and E. coli and the increase of bifidobacteria.

Animals ; Bacteria ; classification ; genetics ; isolation & purification ; Biodiversity ; Chicory ; chemistry ; metabolism ; Cholesterol ; metabolism ; Gastrointestinal Tract ; microbiology ; Humans ; Male ; Microbiota ; Obesity, Abdominal ; metabolism ; microbiology ; Plant Extracts ; metabolism ; Rats ; Rats, Sprague-Dawley ; Triglycerides ; metabolism