Base Sequence ; Conjugation, Genetic ; Drug Resistance, Bacterial ; Electrophoresis, Gel, Pulsed-Field ; Escherichia coli ; drug effects ; enzymology ; Humans ; Klebsiella pneumoniae ; drug effects ; enzymology ; Microbial Sensitivity Tests ; Molecular Sequence Data ; Polymerase Chain Reaction ; Transformation, Bacterial ; beta-Lactamases ; genetics

To develop an analytic method for qualitative and quantitative analysis of dodecatetraenamides A, B in 42 samples of two official species of Asari Radix et Rhizoma( ARR) (37 samples of Asarum heterotropoides var. mandshuricum with different collection time and 5 samples of Asarum sieboldiivar. seoulense). The HPLC-IT-TOF-MS/MS methods for the qualitative and UPLC-PDA methods for the quantitative analysis were established. Dodecatetraenamides A, B were identified by comparing the retention time, UV absorption spectrum and quasi-molecular ion peak [ M + H]+ with the reference compound using HPLC-IT-TOF-MS/MS. The content of dodecatetraenamides A and B in ARR were determined by UPLC-PDA. The separation was successfully carried out on a ACQUITY UPLC BEH C18 (2.1 mm x 100 mm, 1.7 µm) column eluted with mobile phases of water (A) and acetonitrile (B) in gradient program (0-3 min, 35% B; 3-5 min, 35%-36% B; 5-6 min, 36%-43% B; 6 min-11 min 43% B; 11-12 min, 43%-100% B). The column temperature was 45 °C, and the detection wavelength was set at 254 nm. The flow rate was 0.6 mL · min(-1). On one level mass spectrometry scanning, the results showed that the quasi-molecular ion [M + H] + of both dodecatetraenamides A and B were m/z 248.20. The quantitative method with UPLC-PDA has made the baseline separation of the constituents, which were reported as mixtures in the most literatures. The average recovery of dodecatetraenamides A and B were 97.90% and 99.86%, the relative standard deviation were 0.4% and 1.1%, respectively. The contents of dodecatetraenamides A, B in all ARR samples was in the range of 0.11-3.89 and 0.24-6.65 mg · g(-1). Their contents reduced with the extension of storage time. Compared with the samples of 2013, the average content of the two constituents in the samples collected in year 2002-2003 reduced 34% and 36%, respectively (P < 0.05). Compared the A. sieboldii var. seoulense and A. heterotropoides var. mandshuricum with the same collective time and production area, the average contents of the two constituents in latter were up to (1.59 ± 0.75) mg · g(-1) and (2.90 ± 1.17) mg · g(-1), respectively, significantly higher than that in A. sieboldii var. seoulense (dodecatetraenamide A were (0.78 ± 0.52) mg · g(-1), dodecatetraenamide B were (1.69 ± 0.83) mg · g(-1)) (P < 0.05). The content of the dodecatetraenamide A in overground part was in the range of 0.11-0.33 mg · g(-1), dodecatetraenamide B was 0. 24-0.60 mg · g(-1), which were much lower than that of the underground part of ARR (dodecatetraenamide A was in the range of 0.73-3.89 mg · g(-1), dodecatetraenamide B was 2.11-6.24 mg · g(-1)). The method was certified to be simple, accurate and reliable and could be used for qualitative and quantitative analysis of dodecatetraenamide A and B in different species of ARR, also can be used for the comprehensive quality control of traditional Chinese medicine, Asari Radix et Rhizoma.
Amides ; chemistry ; Asarum ; chemistry ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Mass Spectrometry ; Molecular Structure ; Rhizome ; chemistry

The contents of adenosine, gastrodin, 4-hydroxybenzyl alcohol, 4-hydroxybenzaldehyde, parishin and sulfur dioxide residue were compared in differently-processed Gastrodiae Rhizoma to provide the basis for a reasonable processing method of Gastrodiae Rhizoma. The analysis was performed on a Merck Purospher STAR column (4.6 mm x 250 mm, 5 μm) with a mobile phase consisting of methanol and water (containing 0.1% formic acid) under gradient elution at a flow rate of 1.0 mL x min(-1). The eluates were detected at 270 nm, and the column temperature was 35°C. The content of adenosin, gastrodin, 4-hydroxybenzyl alcohol, 4-hydroxy-benzaldehyde and parishin in processing of boiling or sulfur-fumigated were lower than that of in processing of steaming. Furthermore, the sulfur dioxide residue of sulphur-fumigated groups exceed 400 mg x kg(-1). This stable and reliable method will contribute to the quality control of different processed Gastrodiae Rhizoma.
Drug Contamination ; Drugs, Chinese Herbal ; chemistry ; Gastrodia ; chemistry ; Sulfur Dioxide ; analysis ; Technology, Pharmaceutical ; methods

OBJECTIVETo investigate the effect of cytomegalovirus infection on expression of matrix metalloproteinase-2 in human endothelial cells.

METHODSHuman umbilical vein endothelial cells were cultured in vitro. Cells between 3-6 passages were infected with cytomegalovirus for different time. Expression of matrix metalloproteinase-2 mRNA was analyzed by reverse transcription-polymerase chain reaction, matrix metalloproteinase-2 activity was detected by gel zymography.

RESULTSExpression of matrix metalloproteinase-2 mRNA and its activity 6 hours after infection was almost equal to control, and was greatly enhanced 12 and 24 hours after infection.

CONCLUSIONCytomegalovirus infection up-regulates expression of matrix metalloproteinase-2 in human endothelial cells. It might be one of the mechanisms that cytomegalovirus is involved in atherosclerosis.

Cells, Cultured ; Cytomegalovirus ; physiology ; Endothelial Cells ; cytology ; metabolism ; virology ; Gene Expression Regulation, Enzymologic ; Host-Pathogen Interactions ; Humans ; Matrix Metalloproteinase 2 ; biosynthesis ; genetics ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Umbilical Veins ; cytology

Objective To investigate the clinical manifestation, cerebruspinal fluid(CSF) and auxiliary examination findings of adult viral meningitis. Methods 62 adult patients with viral meningitis were retrospectively analyzed. Results Headache occurred in all the 62(100%) patients, fever occurred in 61 (98%) patients, meningeal irritation sign occurred in 48 (77%) patients. The abduction of left eye was limited in one patient. Seizure occurred in 2 patients. The mean duration time was 17 days, 93% patients less than 30 days. The pressure of CSF increased in 80% patients, leukocyte counts increased in 91% patients, protein level increased in 81% patients, chloride level was normal in 35% patients and sfighfly lower in 65% patients, glucose level was normal in 94% patients. 7 patients had positive IgM antibody of Coxackievirus B group both in serum and CSF,one patient had positive IgM antibody of EB virus in CSF. Cranial CT scan had no special findings in all patients.23 patients performed MRI examination, meningeal enhancement occurred in 9 patients. 52% patients had abnormal EEG, mainly increased local or diffuse slow waves. Conclusion Adult viral meningitis was a kind of selflimited disease, chloride level was shghtly lower in more than half patients, meningeal enhancement was detected in MRI in part patients.

To study the chemical constituents of Dioscorea zingiberensis Wright, the EtOH extract of fresh rhizomes of D. zingiberensis was concentrated and partitioned further to produce petroleum ether-, ethylacetate-, n-butanol- and water-soluble fractions. The water-soluble fraction was subjected to column chromatography on macro resin AB-8, and the final products were obtained by repeated reversed-phase ODS and MCI gel CHP 20P column chromatography. Structures of compounds were elucidated by 1H NMR, 13C NMR, 135DEPT, HMQC, HMBC and TOCSY spectroscopic analyses. A new steroidal saponin was isolated, which was identified as (25R)-26-O-(beta-D-glucopyranosyl)-furost-5-en-3 beta, 16, 20, 26-tetraol-22-seco-3-O-beta-D-glucopyranosyl-(1--> 3)-beta-D-glucopyranosyl-(1--> 4)-[alpha-L-rhamnopyranosyl-(1-->2)]-beta-D-glucopyranoside. The compound is a novel skeletally steroidal saponin, named as zingiberenin F (1). It was reported for the first time from D. zingiberensis Wright.
Dioscorea ; chemistry ; Molecular Structure ; Plants, Medicinal ; chemistry ; Rhizome ; chemistry ; Saponins ; chemistry ; isolation & purification

Objective To discuss the clinical effect of percutaneous endoscopic lumbar discectomy(PELD) in the treatment of lumbar disc herniation and calcification.Methods The clinical data of 52 patients with lumbar disc herniation and calcification in orthopedic department of general hospital of Fushun mining bureau from June 2015 to June 2016 were retrospectively reviewed.The data included medical records,out-patient review,telephone follow-up was collected.The VAS,ODI and modified MacNab criteria were used to assess the clinical effects.Results There were significant improvement in VAS and ODI score at 6 week,6 months,1 year after surgery compared with before.According to the modified MacNab criteria,the rate of excellent and good result was 94.3％,the improvement rate was 98.1％.Conclusion PELD is an effective method to treat lumbar disc herniation and calcification,with advantages of less injury,rapid recovery,it is worth popularizing application in clinic.

OBJECTIVETo explain the molecular evidence of revision of taxonomic placement of Peucedanum decursivum based on the nrDNA ITS sequence.

METHODPCR amplification, DNA sequencing and cladistic analysis.

RESULTThe ITS sequences and phylogenetic tree of 5 species of Angelica were and Peucedanum were acquired, in which 5 species were divided into 2 groups, Angelica group and Peucedanum group. P. decursivum was placed in the Angelica group.

CONCLUSIONP. decursivum belongs to genus Angelica. The scientific name of P. decursivum should be revised as A. decursivum. A. decursivum and P. praeruptorum should be used as crude drug respectively.

Angelica ; classification ; genetics ; Apiaceae ; classification ; genetics ; Base Sequence ; DNA, Plant ; genetics ; DNA, Ribosomal Spacer ; genetics ; Molecular Sequence Data ; Phylogeny ; Plant Leaves ; genetics ; Plants, Medicinal ; classification ; genetics

OBJECTIVETo evaluate the efficacy of combined transplantation of neurotrophin-3 and neural stem cells in treatment of hypoxic-ischemic brain injury in rats, and study the possible mechanism.

METHODSNeural stem cells (NSCs) isolated from hippocampi of newborn wistar rats were cultured and identified. The hypoxic-ischemic brain injury models established with 7-day-old wistar rats, and neural stem cells or others were transplantated into the ipsilateral ventricles 7 days later. The 7-day-old rats were randomized into 5 groups: normal group, model group, sham-transplantation group, NSCs transplantation group, and NT-3 combined with NSCs transplantation group. There were 12 rats per group. The functional test and immunohistochemistry were examined 4 weeks later.

RESULTSThe neural stem cells from new-born rats' hippocampi were successfully cultured. It was found that they formed typical neurospheres in suspension, and the majorities of cells expressed nestin, which was the marker for neural stem cells. The rats from combined transplantation group performed significantly better in the ability to study and in memory and the limb function than the rats from NSCs transplantation group (P < 0.05). The rate of neural stem cells differentiating into neurons from combined transplantation group was higher, too (50% vs. 30%).

CONCLUSIONCombined NT-3 and NSCs transplantation could improve the ability to study, memory and the limb function of rats after hypoxic-ischemic brain injury, and improve the rate of NSCs differentiating neurons. Combined NT-3 and NSCs transplantation had better effects on hypoxic-ischemic rats than transplantation of NSCs alone.

Animals ; Animals, Newborn ; Cells, Cultured ; Disease Models, Animal ; Hypoxia-Ischemia, Brain ; therapy ; Neurotrophin 3 ; therapeutic use ; Rats ; Rats, Wistar ; Stem Cell Transplantation

OBJECTIVETo find out the mechanisms of redifferentiation and reversion of malignant human gastric cancer cells induced by ascorbic acid.

METHODSHuman gastric cancer cells grown in the laboratory were used. The Trypan blue dye exclusion method was used to determine the cell doubling time. The electrophoresis rate and colonogenic potential were the indices used to measure the rate of redifferentiation. The content of malondialdehyde (MDA) was measured using the thiobarbituric acid (TBA) method. The activities of superoxide dismutase (SOD), catalase (CAT) and the content of H202 were evaluated by spectrophotography.

RESULTSSix mmol/L ascorbic acid was used as a positive control. Human gastric cancer cells were treated with 75 microm hydrogen peroxide, which alleviated many of the malignant characteristics. For example, the cell surface charge obviously decreased and the electrophoresis rate dropped from 2.21 to 1.10 microm x s(-1) x V(-1) x cm(-1). The colonogenic potential, a measure of cell differentiation, decreased 90.2%. After treatment with ascorbic acid, there was a concentration- and time-dependent increase in hydrogen peroxide (H202) and the activity of superoxide dismutase (SOD). However, the activity of catalase (CAT) resulted in a concentration- and time-dependent decrease. SOD and 3-amino-1,2,4-triazole (AT) exhibited some effects, but there were statistically significant differences between the SOD and AT group and the H202 group.

CONCLUSIONSAscorbic acid induces growth inhibition and redifferentiation of human gastric cancer cells through the production of hydrogen peroxide.

Antioxidants ; pharmacology ; Ascorbic Acid ; pharmacology ; Cell Differentiation ; drug effects ; Humans ; Hydrogen Peroxide ; metabolism ; Stomach Neoplasms ; drug therapy ; metabolism ; pathology ; Tumor Cells, Cultured