OBJECTIVE:To study the characteristics of patent behaviors of Chinese pharmaceutical demonstration enterprises for intellectual property,and to provide reference for the improvement of patent behaviors in Chinese pharmaceutial enterprises. METHODS:After retrieving the applicants(patentees)from China Intellectual Property Right Net on the basis of taking 7 pharma-ceutical demonstration enterprises chosen through appraisal by State Intellectual Property Office of the P.R.C in 2013 as the study samples,empirical study was conducted on their patent behaviors during 1985 and 2013,including the number and type of the pat-ents for which applications were made,international patent classification and legal status. RESULTS:The number of patent applica-tions of the 7 pharmaceutical demonstration enterprises demonstrated a growing trend(an annual increase up to 79.0%). The appli-cations were dominated by the inventive patent(84.9%). Many patents were in an authorized legal status(46.7%). All of these en-terprises paid more attention to the maintenance of the authorized patent. The technology patent applications were mainly about chemical compound,composition,and preparation methods,including the technical proposal containing the active ingredients and properties of the pharmaceutical preparation (A61K),particularly the application of the technical proposal related to the treatment with the preparation (A61P). CONCLUSIONS:All the 7 pharmaceutical demonstration enterprises were aware of patent competi-tion and protection and characterized by a higher proportion of the application for the inventive patent,perfect patent information management institution and high-quality patent applications. Chinese pharmaceutical enterprises can enhance their competitiveness and creativity through adjusting patent behaviors and increasing their awareness of patent competition.

OBJECTIVETo explore the correlation between electronic bronchus mirror and Chinese medical syndrome typing of Mycoplasma pneumonia children.

METHODSTotally 198 Mycoplasma pneumonia children inpatients were assigned to three syndrome types according to Chinese medical syndrome typing and self-formulated typing standards of electronic bronchus mirror, i.e., Fei-qi accumulation of damp and heat syndrome, Fei-qi accumulation of toxicity and heat syndrome, deficient vital qi leading to lingering of pathogen syndrome. The correlation between electronic bronchus mirror and Chinese medical syndrome typing was explored.

RESULTSAs for comparison between electronic bronchus mirror and Chinese medical syndrome typing, Kappa value (K^) was 0.645 and Spearman coefficient correlation (r) was 0.653 (P < 0.01) for Fei-qi accumulation of damp and heat syndrome; K^ was 0.724 and r(s) was 0.727 (P < 0.01) for Fei-qi accumulation of toxicity and heat syndrome; K^ was 0.506 and r(s) was 0.515 (P < 0.01) for deficient vital qi leading to lingering of pathogen syndrome.

CONCLUSIONChinese medical syndrome typing of Mycoplasma pneumonia children was moderately in line with inspection typing under electronic bronchoscope with significant correlation.

Objective To evaluate the prognostic factors in locally advanced non-small-cell lung cancer (LA-NSCLC) for selectively carrying out prophylactic cranial irradiation (PCI).Methods 114 patients with LA-NSCLC between Jun 2006 and Oct 2010 were retrospectively analyzed. Related risk factors and features about brain metastases were analyzed.Results The 2-year incidence rate of brain metastases was 31.58 ％ (36/114),the first brain metastases was 20.18 ％ (23/114),and sole brain metastases was 9.65 ％(11/114),respectively.Variables involved in the equation of binary logistic regression analysis were pathology (OR =5.892) and treatment mode(OR =2.888).The incidence rate of brain metastases in patients of non-squamous carcinoma and single treatment mode was higher than others (P ＜ 0.01) Model fitting is better (P ＞ 0.05).Overall accuracy rate of predicting brain metastases is 67.7 ％.The increased rate of lactate dehydrogenase in the patients with brain metastases or death was 17.54 ％, which was higher than that in the survival patients without brain metastases (P ＜ 0.01).At the same time,the station number and the number of mediastinal lymph node metastases were positively correlated (r =0.716, P ＜ 0.01).The incidence rate of brain metastases or mortality rate was higher in the adenocarcinoma cases than that in the squamous carcinoma cases (P ＜ 0.01,P ＜ 0.05),with more frequent occurrence of mediastinal metastases.The mean diameter of squamous carcinoma and adenocarcinoma were 5.8 cm and 3.9 cm, respectively (P ＜ 0.01).Conclusions The incidence rate of brain metastases was higher in patients with single treatment.Large primary tumors, high lactate dehydrogenase, non-squamous carcinoma, multiple stations, and multiple mediastinal lymph nodes metastases can be regarded as risk factors of brain metastases to perform PCI.

Epiberberine, one of the most important isoquinoline alkaloid in Coptidis Rhizoma, possesses extensive pharmacological activities. In this paper, the liquid chromatography-tandem mass spectrometry (LC-MS/MS) was used to study phase I and phase II metabolites. A Thermo HPLC system (including Surveyor AS, Surveyor LC Pump, Surveyor PDA. USA) was used. The cocktail probe drugs method was imposed to determine the content change of metoprolol, dapsone, phenacetin, chlorzoxazone and tolbutamide simultaneously for evaluating the activity of CYP2D6, CYP3A4, CYP1A2, CYP2E1 and CYP2C9 under different concentrations of epiberberine in rat liver microsomes. The result showed that epiberberine may have phase I and phase II metabolism in the rat liver and two metabolites in phase I and three metabolites in phase II are identified in the temperature incubation system of in vitro liver microsomes. Epiberberine showed significant inhibition on CYP2D6 with IC50 value of 35.22 μmol L(-1), but had no obvious inhibiting effect on the activities of CYP3A4, CYP1A2, CYP2E1 and CYP2C9. The results indicated that epiberberine may be caused drug interactions based on CYP2D6 enzyme. This study aims to provide a reliable experimental basis for its further research and development of epiberberine.
Animals ; Berberine ; analogs & derivatives ; chemistry ; metabolism ; Chromatography, High Pressure Liquid ; Cytochrome P-450 CYP2D6 ; metabolism ; Cytochrome P-450 CYP2D6 Inhibitors ; chemistry ; metabolism ; Drugs, Chinese Herbal ; chemistry ; metabolism ; Male ; Microsomes, Liver ; drug effects ; enzymology ; metabolism ; Molecular Structure ; Rats ; Rats, Sprague-Dawley ; Tandem Mass Spectrometry

To figure out the stability and intestinal bacteria metabolites of rats in vitro of astragaloside IV ( AST), this research was done to explore the stability of AST in the artificial gastric juice. artificial intestinal juice and rat liver homogenate and the metabolism in rat intestinal in vitro. HPLC was used to calculate the remaining rate of AST in biological samples by measuring the content of AST, while metabolites were determined by combining the methods of TLC, HPLC and LC-MS/MS. It turned out that AST was difficult to metabolize in the artificial gastric juice, artificial intestinal juice and rat liver. Also, the metabolic pathway of AST was stepped by deglycosylation. Firstly, AST was converted to its secondary etabolites (6-O-β-D-glucopyranosyl- cycloastragenol, CMG) by removal of xylose moiety at C-3, then transformed into cycloastragenol (CAG) after hydrolytic removal of the glucose moiety at C-6. All the results suggested that the metabolism of AST in vivo occurs mainly in the intestinal by hydrolysis of glycosyl. In conclusion, hydrolysis of intestinal flora is the main reason that AST metabolizes.
Animals ; Bacteria ; metabolism ; Chromatography, High Pressure Liquid ; Drug Stability ; Intestines ; microbiology ; Liver ; metabolism ; Rats ; Rats, Sprague-Dawley ; Saponins ; chemistry ; metabolism ; Tandem Mass Spectrometry ; Triterpenes ; chemistry ; metabolism

Objective Three-dimension (3D) cell matrix adhesion in vivo is fundamentally important for a wide variety of cellular physiological and pathological phenomena, however, the cell-matrix 3D adhesion is hardly observed in vitro. We present the human foreskin fibroblasts (HFF) formed 3D adhesion complexes on the thoroughly acellular human amniotic matrix (TAHAM). Methods TAHAM were produced by suspending digestion with trypsin. The HFF were seeded on 6 well plate, matrigel and TAHAM individually.The light microscope, scanning electronic microscope, immunohistochemistry and immunofluorescence were used to observe the micro-structures and detect the type Ⅰ , Ⅲ, Ⅳ, Ⅵ collagen, laminin, fibronectin, TGF-β1, TGF-β2, FGF of the TAHAM. Phase contrast microscope was engaged to observe the morphology of HFF. The time-lapse CCD and the trace analysis software were employed to prescribe the cell migration. The 3D adhesion foci were identified by the laser confocal microscope. The strain of the TAHAM was tested by the universal mechanical testing instrument. Results The fibers of the TAHAM were intact, type Ⅰ , Ⅲ,Ⅳ, Ⅵ collagen, laminin, fibronectin were positive, TGF-β1, TGF-β2, FGF were negative. HFF had a bipolar extension to form multilayer cell clusters networks and grew into the matrix. All of the seeded cells survived three weeks under regular culture without transfer. On TAHAM, HFF moved in a straight line with a speed of 12 μm/h. α5 integrin (green), paxillin (red) and fibronectin (blue) co localized to form 3D adhesion complexes (white). Conclusion The main molecular components and biomechanical properties is preserved in TAHAM. HFF forms 3D adhesion complexes on TAHAM. Cell morphology and migration of HFF on TAHAM correspond to that under 3D adhesion behavior.

In this paper, the chloroplast psbK-psbI intergenic spacers of 18 species of Dendrobium and their adulterants were amplified and sequenced, and then the sequence characteristics were analyzed. The sequence lengths of chloroplast psbK-psbI regions of Dendrobium ranged from 474 to 513 bp and the GC contents were 25.4%-27.6%. The variable sites were 71 while the informative sites were 46. The inter-specific genetic distances calculated by Kimura 2-parameter (K2P) of Dendrobium were 0.006 1-0.058 1, with an average of 0.028 4. The K2P genetic distances between Dendrobium species and Bulbophyllum odoratissimum were 0.093 2-0.120 4. The NJ tree showed that the Dendrobium species can be easily differentiated from each other and 6 samples of the inspected Dendrobium species were identified successfully through sequencing the psbK-psbI intergenic spacer. Therefore, the chloroplast psbK-psbI intergenic spacer can be used as a candidate marker to identify Dendrobium species and its adulterants.
Chloroplasts ; DNA, Chloroplast ; genetics ; DNA, Plant ; genetics ; DNA, Ribosomal Spacer ; genetics ; Dendrobium ; classification ; genetics ; Plants, Medicinal ; classification ; genetics

In order to identify Peucedani Radix, Peucedani Decursivi Radix and their adulterants, the internal transcribed spacer 2 (ITS2) regions of Peucedani Radix, Peucedani Decursivi Radix and their adulterants were amplified and bidirectionally sequenced based on the Principles for Molecular Identification of Traditional Chinese Materia Medica Using DNA Barcoding, which has been promulgated by Chinese Pharmacopoeia Commission. Sequences were analyzed and assembled by Codon Code Aligner V3. 7.1. The relevant data were analyzed by MEGA 5. 0. Species identification analyses were performed by using the nearest distance methods and neighbor-joining (NJ) methods. The result showed that the ITS2 sequence lengths of Peucedani Radix were 229-230 bp and the average intra-specific genetic distances were 0.005. The ITS2 sequence lengths of Peucedani Decursivi Radix were 227 bp and the sequences contained no variation site. The average inter-specific K2P genetic distance of Peucedani Radix, Peucedani Decursivi Radix and their adulterants species were 0.044 and 0.065 respectively. The minimum inter-specific divergence is larger than the maximum intra-specific divergence of Peucedani Decursivi Radix. The nearest distance methods and NJ trees results indicated that Peucedani Radix, Peucedani Decursivi Radix and their adulterants species could be identification clearly. The ITS2 regions can stably and accurately distinguish Peucedani Radix, Peucedani Decursivi Radix and their adulterants.
Apiaceae ; classification ; genetics ; DNA Barcoding, Taxonomic ; methods ; DNA, Ribosomal Spacer ; Drug Contamination

OBJECTIVETo identify the Cynomorii Herba and its analogues species using DNA barcoding technique.

METHODTotal genomic DNA extracted from all materials using the DNA extraction kit. The internal transcribed spacer 2 (ITS2) regions were amplified using polymerase chain reaction (PCR), and purified PCR products were sequenced bi-directionally. Sequence assembly and consensus sequence generation were performed using the CodonCode Aligner 3.7.1. The Kimura 2-Parameter (K2P) distances and GC content were computed using MEGA 5. 0. Species identification analyses were conducted through the species identification system for traditional Chinese medicine and neighbor-joining (NJ) trees.

RESULTThe ITS2 sequence lengths of Cynomorii Herba were 229 bp. The average intra-specific genetic distances of Cynomorii Herba were 0.003. The average inter-specific genetic distances between Cynomorii Herba and its adulterants species were 0.760. The results showed that the minimum inter-specific divergence is larger than the maximum intra-specific divergence. The species identification system for traditional Chinese medicine and NJ trees results indicated that Cynomorii Herba and its adulterants species can be easily identification.

CONCLUSIONThe ITS2 region is an efficient barcode for identification of Cynomorii Herba, which provide a new technique to ensure clinical safety in utilization of traditional Chinese medicine.

Aging ; Animals ; Drugs, Chinese Herbal ; pharmacology ; Glutathione Peroxidase ; metabolism ; Male ; Malondialdehyde ; metabolism ; Rats ; Rats, Wistar ; Superoxide Dismutase ; metabolism ; Testis ; drug effects ; metabolism