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

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

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

OBJECTIVETo investigate the clinical manifestation, cerebrospinal fluid (CSF) and auxiliary examination findings of adult viral meningitis.

METHODS62 adult patients with viral meningitis were retrospectively analyzed.

RESULTSHeadache 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 slightly 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.

CONCLUSIONAdult viral meningitis was a kind of self-limited disease, chloride level was slightly lower in more than half patients, meningeal enhancement was detected in MRI in part patients.

Adolescent ; Adult ; Aged ; Antibodies, Viral ; cerebrospinal fluid ; Brain ; diagnostic imaging ; Enterovirus B, Human ; immunology ; Female ; Humans ; Immunoglobulin M ; cerebrospinal fluid ; Magnetic Resonance Imaging ; Male ; Meningitis, Viral ; cerebrospinal fluid ; diagnosis ; diagnostic imaging ; metabolism ; Middle Aged ; Radiography ; Retrospective Studies ; Young Adult

OBJECTIVEThe goal of this study was to investigate whether murine cytomegalovirus (MCMV) is able to exacerbate the atherosclerotic process in apolipoprotein E knockout (apoE -/-) mice, and the effect of fluvastatin on the atherogenesis.

METHODSThe apoE-/- mice kept on a west diet were given low dosage of MCMV. At 14,18 and 24 weeks post infection, AS lesion were measured on aorta. The fluvastatin was administered, and AS lesion were measured accordingly above.

RESULTSWe observed that in the chronic phase of the infection, AS lesion area was significantly increased. MCMV gB mRNA was not amplified by real-time PCR from the arterial wall. The IgG antibody level of MCMV in blood plasma and the content of virus DNA in salivary gland were not correlated with AS lesions. After the administration of fluvastatin, there was no significant difference of AS lesions between MCMV infected group and mock-infected group.

CONCLUSIONMCMV may aggravate the AS lesion in apoE -/- mice in the chronic phase of infection, and promote more severe type of AS lesions. But it might not be the direct effects of mechanism of MCMV on the local lesion of AS. Fluvastatin could meliorate the progression of AS after MCMV infection, but this was not accomplished by decreasing MCMV duplication.

Animals ; Aorta ; drug effects ; Apolipoproteins E ; deficiency ; genetics ; Atherosclerosis ; blood ; drug therapy ; genetics ; virology ; Fatty Acids, Monounsaturated ; pharmacology ; Herpesviridae Infections ; blood ; drug therapy ; virology ; Immunoglobulin G ; blood ; Indoles ; pharmacology ; Male ; Mice ; Mice, Knockout ; Muromegalovirus ; genetics

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

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 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

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.