Several kinds of column chromatography method were used to investigate the chemical constituents of the leaves of Lophatherum gracile. The structures of the isolated compounds were identified based on their physicochemical properties and spectral data. A new flavone C-glycoside was isolated and its structure was identified as 3'-methoxyl-luteolin 6-C-beta-D-galactopyranosiduronic acid (1 --> 2) -alpha-L-arabinopyranoside (1).
Antiviral Agents ; chemistry ; isolation & purification ; Flavones ; chemistry ; Glycosides ; chemistry ; isolation & purification ; Hydrolysis ; Plant Leaves ; chemistry ; Poaceae ; chemistry

AIM:To investigate the effects and mechanism of anti-vascular endothelial growth factor ( VEGF) assisted pars plana vitrectomy ( PPV) in the treatment of proliferative diabetic retinopathy ( PDR) . ·METHODS: A total of 92 patients ( 92 eyes ) with PDR treated by PPV were divided into the simple PPV group (41 patients with 41 affected eyes) and the combined treatment group ( 51 patients with 51 affected eyes ) according to whether the patient underwent intravitreal injection of Ranibizumab ( IVR) . The combined treatment group was treated with IVR at 5-7d before PPV. The surgical time, times of electrocoagulation, silicone oil filling rate, the incidence of postoperative complications, LogMAR BCVA of affected eyes, levels of VEGF and pigment epithelium derived factor ( PEDF ) in aqueous humor and vitreous body were compared between the two groups. ·RESULTS:The surgical time was shorter, the times of electrocoagulation was less, the silicone oil filling rate and the incidence rates of iatrogenic retinal hole and vitreous body hematocele were lower in the combined treatment group than in the simple PPV group (P<0. 05). Levels of VEGF and PEDF in aqueous humor and vitreous body of the combined treatment group during PPV were lower than those in the simple PPV group (P<0. 05). The LogMAR BCVA of the affected eyes of the combined treatment group in 3mo after surgery was better than that of the simple PPV group (P<0. 05). ·CONCLUSION:IVR combined with PPV can reduce the perioperative levels of VEGF and PEDF, reduce the times of electrocoagulation and the incidence of iatrogenic retinal hole and vitreous body hematocele, and improve the visual acuity of patients with PDR.

Response Surface Methodology was applied to optimize the culture components for lipopeptide production by Bacillus natto TK-1. In the first step, two level factorial design of Plackett-Burman was used to evaluate the influence of six related factors. It showed that three factors playing the important roles in the medium, including peptone, yeast extract powder and CaCl_2. The path of steepest ascent was used to approach the optimal region of the fermentation conditions subsequently. In the third step, the concentrations of those three main factors were further optimized by using Box-Behnken and Response Surface Analysis. By solving the quadratic regression model equation, the optimal concentrations of the variables were determined as: peptone 1.73%, yeast extract powder 0.063 %, CaCl_2 1.385?10-4mol/L. Under the optimal culture conditions, the diameter of haemolysis zone increased 29.3 % than before. HPLC analysis showed the precise production of lipopeptide was 30.2% higher than preliminary culture. Furthermore, at three batches cultivation, the experiment values under the optimal conditions agreed with the predictive values. It showed that Response Surface Methodology was proper and a good choice for optimization.

1H NMR metabonomics approach was used to reveal the chemical difference of urine between patients with Xiao-Chaihu Tang syndrome (XCHTS) and healthy participants (HP). The partial least square method was used to establish a model to distinguish the patients with Xiao-Chaihu-Tang syndrome from the healthy controls. Thirty-four endogenous metabolites were identified in the 1H NMR spectrum, and orthogonal partial least squares discriminant analysis showed the urine of patients with Xiao-Chaihu Tang syndrome and healthy participants could be separated clearly. It is indicated that the metabolic profiling of patients with Xiao-Chaihu Tang syndrome was changed obviously. Fifteen metabolites were found by S-pot of OPLS-DA and VIP value. The contents of leucine, formic acid, glycine, hippuric acid and uracil increased in the urine of patients, while threonine, 2-hydroxyisobutyrate, acetamide, 2-oxoglutarate, citric acid, dimethylamine, malonic acid, betaine, trimethylamine oxide, phenylacetyl glycine, and uridine decreased. These metabolites involve the intestinal microbial balance, energy metabolism and amino acid metabolism pathways, which is related with the major symptom of Xiao-Chaihu Tang syndrome. The patients with Xiao-Chaihu Tang syndrome could be identified and predicted correctly using the established partial least squares model. This study could be served as the basis for the accurate diagnostic and reasonable administration of Xiao-Chaihu-Tang syndrome.
Humans ; Least-Squares Analysis ; Medicine, Chinese Traditional ; Metabolome ; Metabolomics ; Proton Magnetic Resonance Spectroscopy ; Syndrome ; Urinalysis

Objective: To investigate the function and mechanism of phospholipase Cγ1 (PLCγ1) in cell-matrix adhesion in colorectal cancer. Methods: Highly metastatic colorectal cancer cell line LoVo and lowly metastatic colorectal cancer cell line SW480 were subjected to cell-matrix adhesion assay. U73122 (a specific inhibitor of PLC) and pyrrolidine dithiocarbamate (PDTC) (an inhibitor of NF-κB) were used to study the effect of PLCγ1 and NF-κB on cell-matrix adhesion. Furthermore, Western blot and gel electrophoresis mobility shift assay (EMSA) were performed to detect the mechanism of PLCγ1 in colorectal cancer cell adhesion to matrix. Results: Inhibition of PLCγ1 or NF-κB resulted in reduction of cell-matrix adhesion in a dose-dependent manner in LoVo cells(P<0.05), but had no marked effect on SW480 cells. Western blot analysis showed that epidermal growth factor (EGF) stimulated the phosphorylation of PLCγ1 in LoVo. The results of EMSA indicated that inhibition of PLCγ1 signaling pathway also down-regulated the activity of NF-κB while EGF reversed the function. Conclusion:These data suggest that PLCγ1 plays a pivotal role in the EGF-induced cell-matrix adhesion of highly metastatic colorectal cancer cells and that NF-κB is also functional in this signaling pathway.

ORF1 and ORF2 gene of porcine circovirus type 2 were cloned by PCR with the specific primers designed according to genome of PCV2 (AY035820). Following extraction and digestion, PCR products were subsequently inserted into universal transfer vector plECMV (deleted partial gE and gI of pseudorabies virus) to generate recombinant transfer plasmid pIEORF1-ORF2. The genomic DNA of PRV TK-/gE- /LacZ+ strain and pIEORF1-ORF2 were co-transfected into IBRS-2 cells with lipofectin, and recombinant virus TK- /gE- /gI- /ORF1-ORF2+ was selected by PCR with ORF1 gene and ORF2 gene primers respectively. The recombinant virus was analyzed with Southern blotting and Western blotting. The results indicated that ORF1 and ORF2 gene of PCV2 had been inserted into the genome of TK- /gE- /LacZ+ strain and the expressed ORF1-ORF2 fusion protein could react with PCV2 positive sera. Result of virus titers detection showed the insertion of ORF1 and ORF2 gene did not influence propagation of recombinant virus.
Animals ; Cell Line ; Circovirus ; classification ; genetics ; Gene Transfer Techniques ; Genes, Viral ; Herpesvirus 1, Suid ; genetics ; Open Reading Frames ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; Recombination, Genetic ; Swine

This paper is aimed to develop rapid, sensitive and convenient HPLC-MS/MS methods for the quantification of levosimendan and its metabolites OR-1855 and OR-1896 in human plasma. According to the different natures of the compounds, two sets of liquid chromatography and ionization modes were used for determination the concentration of levosimendan and its metabolites OR-1855 and OR-1896 in human plasma, separately. Following protein precipitation with methanol, the levosimendan and internal standard (rosuvastatin) were separated on a Capcell MG III C18 column (35 mm x 2.0 mm ID, 3 microm) with the mobile phase consisted of methanol-15 mmol x L(-1) ammonium acetate-formic acid (55: 45: 0.02, v/v/v). A tandem mass spectrometer equipped with electrospray ionization source was used as the detector and operated in the negative ion mode. Its metabolites OR-1855, OR-1896 and internal standard doxofylline were extracted from plasma by liquid-liquid extraction with ethyl acetate. Chromatographic separation was performed on a Zorbax Extend C18 column (150 mm x 4.6 mm ID, 5 microm) with the mobile phase consisted of methanol-15 mmol x L(-1) ammonium acetate-formic acid (65 :35 :0.1, v/v/v). A tandem mass spectrometer equipped with electrospray ionization source was used as the detector and operated at the positive ion mode. The linear concentration ranges of the calibration curves for levosimendan and OR-1855 and OR-1896 were 0.10-50.0 ng x mL(-1), 0.20-100 ng x mL(-1), 0.20-100 ng x mL(-1), respectively. The lower limits of quantification of levosimendan and OR-1855 and OR-1896 were 0.10 ng x mL(-1), 0.20 ng x mL(-1), 0.20 ng x mL(-1), respectively. The methods proved to be sensitive, simple and rapid, and suitable for the pharmacokinetic study of levosimendan injection.
Acetamides ; blood ; Cardiotonic Agents ; blood ; metabolism ; Chromatography, High Pressure Liquid ; methods ; Humans ; Hydrazones ; blood ; metabolism ; Male ; Pyridazines ; blood ; metabolism ; Spectrometry, Mass, Electrospray Ionization ; methods ; Tandem Mass Spectrometry ; methods

The essence of endogenous turbidity in Chinese medicine (CM) is different from cream, fat, phlegm, retention, damp, toxicity, and stasis. Along with the development of modern scientific technologies and biology, researches on the essence of endogenous turbidity should keep pace with the time. Its material bases should be defined and new connotation endowed at the microscopic level. The essence of turbidity lies in abnormal functions of zang-fu organs. Sugar, fat, protein, and other nutrient substances cannot be properly decomposed, but into semi-finished products or intermediate metabolites. They are inactive and cannot participate in normal material syntheses and decomposition. They cannot be transformed to energy metabolism, but also cannot be synthesized as executive functioning of active proteins. If they cannot be degraded by autophagy-lysosome or ubiquitin-prosome into glucose, fatty acids, amino acids, and other basic nutrients to be used again, they will accumulate inside the human body and become endogenous turbidity. Therefore, endogenous turbidity is different from final metabolites such as urea, carbon dioxide, etc., which can transform vital qi. How to improve the function of zang-fu organs, enhance its degradation by autophagy-lysosome or ubiquitin-prosome is of great significance in normal operating of zang-fu organs and preventing the emergence and progress of related diseases.
Autophagy ; Humans ; Medicine, Chinese Traditional ; Proteasome Endopeptidase Complex

OBJECTIVETo analyse the clinical outcome and the prognostic factor of childhood acute myeloid leukemia (AML).

METHODSDisease-free survival (DFS), event-free survival (EFS) and overall survival (OS) rates were estimated by Kaplan-Meier method and prognostic factors were evaluated by Cox regression with SPSS in 141 childhood AML in our hospital from August 1995 to July 2004. The patients were divided into 2 groups: acute promyelocytic leukemia (APL) as group A and AML other than APL as group B.

RESULTSOf the 90 group B patients, 54.4% (49/90) achieved complete remission (CR) after one course chemotherapy , with a total CR rate of 76.7%. The cumulative 5 year DFS and OS rate for group B patients were (28.4 +/- 9.0)% and (35.5 +/- 6.3)%, the 51 group A patients were (94.3 +/- 4.0)% and (81.4 +/- 5.7)%, and for total 141 AML patients were (56.9 +/- 6.3)% and (53.3 +/- 4.8)% respectively. Multivariate analysis demonstrated that higher bone marrow blast cell percentage at diagnosis, CR after more than one course of chemotherapy and less than six courses of consolidation chemotherapy were risk prognostic factors in childhood AML other than APL (P < 0.05).

CONCLUSIONThe prognosis of childhood APL is better, while of childhood t(8;21) AML is no better than other FAB subtypes.

Adolescent ; Bone Marrow Cells ; cytology ; Child ; Child, Preschool ; Disease-Free Survival ; Female ; Humans ; Infant ; Leukemia, Myeloid, Acute ; mortality ; therapy ; Male ; Prognosis ; Regression Analysis ; Retrospective Studies ; Treatment Outcome

Objective To conduct a microbiological and parasitological investigation of experimental minipigs in Guangdong province. Methods Four major experimental minipig production units in Guangdong province were included in this investigation. Samples were taken from a total of 154 pigs of 4 brreds, i. e. , Bama minipigs, Juema minipigs, Tibet minipigs and Wuzhishan minipigs. Pig fur, scales, serum, rectal swabs and feces samples were collected for detection of 20 pathogens. The data were analyzed and compared among the production units and breeds. Results Mixed infections were detected in all the four institutions. The infection rates of 7 pathogens were rather high: Streptococcus suis type 2 (50. 7%), Actinobacillus pleuropneumoniae (40. 3%), Mycoplasma hyopneumoniae (100%), Japanese encephalitis virus (41. 3%), porcine circovirus type 2 (74. 8%), porcine transmissible gastroenteritis virus (73. 8%),gastroenteritis virus (44. 7%). Porcine parvovirus (26. 0%), pseudorabies virus(15. 6%) and intestinal worms (3. 2%) were also detected in some animals. The immune qualified rates of classical swine fever virus (62. 8%) and foot-and-mouth disease virus (35. 8%) were rather low. The immune qualified rate of pseudorabies virus was as high as 98. 4%. Besides, Salmonella, Brucella, swine dysentery snake like spirochetes, dermatophytes, influenza virus. Toxoplasma gondii, ectoparasites, and coccidia were not detected. Conclusions The results of this investigation indicate that epidemiological quality control of pathogens in experimental minipigs and efforts to establish high grade minipig population in Guangdong province remain to be strengthened. Our study also provides a basis for revision of local and even national standards for experimental minipigs.