Reference materials containing mixed degradation products of amoxicillin and ampicillin were developed after optimization of preparation processes. The target impurities were obtained by controlled stress testing, and each major component was identified with HPLC-MS and compared with single traceable reference standard each. The developed reference materials were applied to system suitability test for verifying HPLC system performed in accordance with set forth in China Pharmacopeia and identification of major impurities in samples based on retention and spectra information, which have advantages over the methods put forth in foreign pharmacopoeias. The development and application of the reference materials offer an effective way for rapid identification of impurities in chromatograms, and provide references for analyzing source of impurities and evaluation of drug quality.
Amoxicillin ; chemistry ; Ampicillin ; chemistry ; China ; Chromatography, High Pressure Liquid ; Drug Contamination ; Mass Spectrometry ; Reference Standards

Objective To establish the amplified luminescent proximity homogeneous assay(AlphaLISA) for the detection of Sendai virus.Methods The antigen concentration,serum concentration and the donor beads/acceptor beads ratio used in the AlphaLISA method were optimumized by the phalanx experiments, then the antibodies of Sendai Virus in 40 rat sera were detected by the established AlphaLISA method and ELISA detection method.The results were compared and the differentia between the two methods was confirmed by IFA.Results The optimum concentration of SV bio-peptide in AlphaLISA assay was 250 nmol/L, the best proportion of donor beads/acceptor beads ratio was 1 ∶1, using the concentration of 20 μg/mL and the serum dilution was 1∶10000.7 of the 40 rat sera were detected SV positive by ELISA, the positive rate was 17.5%, 8 of the 40 rat sera were determined SV positive by AlphaLISA, and the positive rate was 20.0%, the AlphaLISA positive serum was confirmed by IFA.Conclusions We preliminary established the Amplified Luminescent Proximity Homogeneous Assay(AlphaLISA) for the detection of Sendai virus.The sensitivity of the method is comparable to classical ELISA method, but this method use less serum samples and without washing steps.The method has some advantages in degeneracy and accuracy.

Animals ; Apolipoproteins ; blood ; Cattle ; Colostrum ; Esterases ; blood ; Female ; Insulin-Like Growth Factor I ; Lipids ; blood ; Nephrotic Syndrome ; blood ; Pregnancy ; Rats ; Rats, Sprague-Dawley

Mountain cultivated ginseng (MCG), also called "Lin-Xia-Shan-Shen", is intentionally seeded and grows in mountainous forest but without any artificial management, which belongs to the herbaceous plants classified in the Araliaceae family and the genus Panax L. MCG has high medicinal value and wide range of pharmacological effects, which had been reported with many kinds of effective components, such as saponins, volatile oil, polysaccharides, protein, and inorganic elements. There is a phenomenon that something regards as real, shoddy, varying of quality in MCG market. Currently, a lot of measures are applied to the identification and evaluation of MCG. This paper summarizes the progress in study on the chemical constituents, the identification and evaluation of MCG, and provides the references for the further development, utilization, and quality control of MCG.

OBJECTIVETo explore the surgical method and its clinical effects of minimally invasive osteosynthesis on the treatment of complex ankle fractures.

METHODSFrom January 2007 to December 2011, 53 patients with complex ankle fractures were treated with minimally invasive osteosynthesis. There were 31 males and 22 females, with an average age of 38.2 years old (ranged, 18 to 65). According to the system of Lauge-Hansen, 32 fractures were supination external rotation injury (grade WV), 13 fractures were pronation external rotation (grade III or IV), 5 fractures were pronation abduction (grade III); and 3 fractures can not be classified due to serious comminution fracture of fibula. According to the system of Denis-Weber, there were 4 cases with type A, 34 cases with type B and 15 cases with type C. Seven cases were open fractures. The duration from injuries to operation ranged from 2 hours to 14 days with an average of 5 days. The sequence of reduction and fixation of ankle fractures was firstly posterior malleolus, then medial malleolus and lateral malleolus, and inferior tibiofibular syndesmosis lastly. The fractures of posterior malleolus were reduced and fixed through anterior ankle approaches; the fractures of medial and lateral malleolus were percutaneously fixed with bolts or blade plate or tensile force band; and inferior tibiofibular syndesmosis were firmly fixed if necessary. Baird-Jackson scoring system was used to evaluate clinical effects.

RESULTSForty-eight patients were followed up from 10 to 36 months with an average of 13 months. The fractures got healing with an average time of 12 weeks (ranged, 10 to 18). According to the Baird-Jackson scoring system, the mean score of ankle function was 94.7 +/- 4.2, and 28 cases obtained excellent results, 15 good, 3 fair and 2 poor. One case experienced superficial infections and was cured by changing dressings, 2 cases experienced fixed syndesmosis screw breakage.

CONCLUSIONThe surgical method of minimally invasive osteosynthesis can ensure the anatomical join restoration, protect the blood supply of fracture end, rebuild the function of ankle joint, obtain satisfactory clinical results in treating complex ankle fractures.

Adolescent ; Adult ; Aged ; Ankle Fractures ; Ankle Joint ; physiopathology ; surgery ; Female ; Fracture Fixation, Internal ; methods ; Humans ; Male ; Middle Aged ; Minimally Invasive Surgical Procedures ; methods

Inflammatory bowel disease (IBD) is a chronic, repeated intestinal inflammatory disease. Clinically commonly used therapeutic drugs have some disadvantages, such as poor efficacy and many adverse reactions after long-term application. Although new biological therapies such as anti-tumor necrosis factor agents, overcome common adverse reactions, also have problems such as high price, difficult storage, drug resistance and recurrence after application. In recent years, many new therapeutic methods for inflammatory bowel disease have emerged, for example, modulators that inhibit lymphocyte migration (integrin inhibitors and sphingosine 1-phosphate receptor agonists) have been introduced into the clinical treatment of inflammatory bowel disease, inflammatory cytokine inhibitors (interleukin-23 inhibitors, Janus kinase inhibitors, phosphodiesterase inhibitors, etc.) and inhibitors targeting fibrosis and intestinal tissue degradation and remodeling (matrix metalloproteinase inhibitors) are also being evaluated in clinical trials of IBD. Based on the mechanisms of action, this paper intends to outline the current mainstream IBD therapies and some emerging drugs, and briefly introduce their targets to provide reference for IBD drug design and development.

OBJECTIVETo explore method of recombinant gene lentivirus containing LIM mineralization protein-1 (LMP-1) in transfecting bone marrow mesenchymal stem cells (BMSC), and to observe the effect of gene LMP-1 on proliferation effect and expression of BMSC.

METHODSSix clean SD rats aged 4 weeks were selected, bone marrow mesenchymal stem cells were extracted under sterile conditions and cultured to the third generation, then divided into three groups:control group (the third generation of BMSC), lentiviral vector transfection group (PGC-FU-GFP and Polybrene were injected into the third generation of BMSC) and recombinant gene transfection group (PGC-FU-LMP-1-GFP and Polybrene transfection were injected into the third generation of BMSC). After 48 hours' transfection, fluorescent expression were detected under immuno-fluorescence microscopy; lentiviral transfection efficiency were detected by flow cytometry; effect of lentiviral transfection on BMSC were evaluated by MTT; gene expression of transfected cells were determined by Western Blot.

RESULTS1) The third generation of BMSC was cultured successfully,and transfected with MOI:100. After 48 hours, green fluorescent expression were detected and transfection efficiency was 67% under immuno-fluorescence microscopy; 2) Compared to control group, there were no statistical differences between control group and other two groups; 3) Western blot teast showed that 72KDa specific band was observed in recombinant gene transfection group and its size was similar to LMP-1 fusion protein (50 kDa+28 kDa=78 kDa).

CONCLUSIONThere is no effect of recombinant gene lentivirus containing LIM on BMSC, and can effectively influence the expression of LMP-1.

Adaptor Proteins, Signal Transducing ; genetics ; metabolism ; Animals ; Cell Proliferation ; Cells, Cultured ; Cytoskeletal Proteins ; genetics ; metabolism ; Female ; Genetic Therapy ; Genetic Vectors ; genetics ; metabolism ; Humans ; LIM Domain Proteins ; genetics ; metabolism ; Lentivirus ; genetics ; metabolism ; Male ; Mesenchymal Stromal Cells ; cytology ; metabolism ; virology ; Osteoporosis ; genetics ; physiopathology ; therapy ; Rats ; Rats, Sprague-Dawley

Objective To establish an indirect immunofluorescence assay for detection of murine norovirus ( MNV) .Methods Mouse leukaemic monocyte macrophage cell line RAW 264.7 cells were infected with MNV-1 and cultured for 36 hours to collect the virus and uninfected cells , and to make antigen glass slides .BALB/c mice were gavaged with MNV-1 (107 TCID50) and infected sera were collected as positive control .The serum was 1:10 diluted and used for measuring MNV antibody by immunofluorescence assay ( IFA ) .80 serum samples were tested using the two methods , IFA and ELISA, and the discrepant samples were validated by Western blotting .Results RAW264.7 cells were infected with MNV-1 for 36-48 h, showing an infection rate of 60% of the cells, and the cells infected for 36 h were preferred.IFA method was used to detect the serum with MNV-1 infection and showed that the antibody content was gradually increased at one week after infection , reaching a maximum antibody concentration at 4 weeks after infection , and maintained a stable level later .The mouse serum at four weeks after MNV-1infection was used as positive quality control . Among the 80 serum samples , 27 positive and 53 negative cases were detected by IFA method , and 32 positive and 48 negative cases were detected by ELISA .The five discrepant samples were verified by Western blotting , resulted in 3 positive and 2 negative cases . The coincidence rate of IFA was 96.0% and that of ELISA methods was 97.5%. Conclusions Basically, immunofluorescence assay can be used to detect the MNV-1 infection in mice, although false negative result may occur occasionally .IFA and ELISA detection can be selected as initial screening measures , and use Western blot assay to verify the discrepant samples .

Based on the 2 x 2 contingency table, by using multi-species relevance (variance ratio, VR), chi2-test, Ochiai index, Dice index, Jaccard index, t-test of v/x and F-test of Morisita, s index, the interspecific relationships and the spatial distribution pattern between 20 dominants in Kangding Zheduo Mountain of Sichuan province were studied. The results indicated that the interspecific association between dominants of Sinopodophyllum hexandrum community in this area did not show significant association, which suggested that the S. hexandrum community was in mature stage, and showed stronger independency, among total 190 pairs in 20 dominant species, 2 species pairs exhibited extremely significantly positive association, 12 species pairs showed significantly positive association, 6 species pairs exhibited significantly negative association and there were no pairs showed extremely significantly negative association. S. hexandrum in community did not show significant association, which indicates they are independent in community, the spatial distribution pattern of S. hexandrum is characterized by random distribution.
Berberidaceae ; classification ; growth & development ; Biodiversity ; China ; Ecosystem ; Endangered Species ; Plants, Medicinal ; classification ; growth & development

OBJECTIVETo observe the effects of bitumen fume on neurotransmitter and ultrastructure of mice brain and to investigate the toxicity of bitumen fume on nerve system of mice brain.

METHODSThe experimental mice were forced to inhale the bitumen fume at different exposure level and in different time periods. The contents of the three transmitters dopamine (DA), norepinephrine (NE), 5-hydroxytryptamine (5-HT) in mice brain were measured by the fluorescence meanwhile ultrastructure of mice brain was observed by electronic microscope. The ultrastructure of mice brain was observed under transmission electron microscopy.

RESULTSThe contents of DA, NE and 5-HT in all groups decreased with the increasing of dose and prolonging of time (after 8 week, with the increasing of exposure lever, the content of DA, NE, 5-HT was respectively 2.194, 2.190, 2.181, 2.178 microg/g and 1.148, 1.138, 1.135 and 1.407, 1.403, 1.395 microg), but the results did not show significant differences. The structure of the mitochondria changes included the swollen mitochondria, chromatin margination, pyknosis and apoptosis in neuro cells and the processes of swollen astrocyte cells.

CONCLUSIONThe bitumen fume could induce changes of the ultrastructure of mice brain and affect the contents of neurotransmitter of mice brain.

Animals ; Brain ; metabolism ; ultrastructure ; Female ; Hydrocarbons ; toxicity ; Male ; Mice ; Mice, Inbred Strains ; Neurotransmitter Agents ; analysis