The HPLC fingerprint differences ofRadix Platycodonis from different origins were studied to provide references for their quality control and production. The total platycodins were purified by DB101 macroporous resin. HPLC-ELSD fingerprints of the total platycodins for 39 batches ofRadix Platycodonis samples from 9 provinces were performed on an Agilent HC-C8 column (4.6 mm x 250 mm, 5μm) with gradient elution. The mobile phase was acetonitrile-0.5% acetic acid. The injection volume was 6μL. The flow rate was 0.5 mL·min-1. The temperature of drift tube was set at 90℃. And the gas flow (N2) was set at 1.2 mL·min-1. The results showed that there were large differences in the quality ofRadix Platycodonis from different origins with the common fingerprints of 6 batches of samples fromChifeng in Inner Mongolia as references. The quality ofRadix Platycodonis was closely related to the seeds, the ecological environment, the way of drying and storing and so on. It was concluded that it was important to strengthen the provenance base construction, standardization of the seeds, reasonable formulation of the regionalization, and standardization of the production processing for the cultivation and production ofRadix Platycodonis.

The microbial transglutamunase (MTG) gene was amplified from the genomic DNA of Streptoverticillium mobaraensea by using PCR and inserted into pET vector to construct the expression plasmid called pET-MTG. The pET-MTG was transfected into E. coli (Rosetta DE3) and the MTG protein was found to be highly expressed as inclusion bodies. The inclusion bodies were isolated and subjected to denaturation and re-naturation, followed by strong cation ion-exchange chromatography to purify the expressed MTG. The specific activity of purified MTG was close to that of native MTG. Taken together, this study might provide a base for the industrial production of microbial transglutaminase.
Bacterial Proteins ; genetics ; metabolism ; Escherichia coli ; genetics ; metabolism ; Genes, Bacterial ; Inclusion Bodies ; enzymology ; Recombinant Proteins ; genetics ; metabolism ; Streptomycetaceae ; enzymology ; genetics ; Transglutaminases ; genetics ; metabolism

OBJECTIVETo research and appraise the method and long-term effects of trigeminal neuralgia by the operation of retrogasserian combing.

METHODSFrom 1994 to 2000, fifty cases of trigeminal neuralgia were treated through posterior fossa microsurgery, 30 of which received both microvascular decompression and retrogasserian combing while 20 only by retrogasserian combing.

RESULTSOf the 50 cases, forty of trigeminal nerves were found to be compressed by arteries, while 28 of them were the superior cerebellar artery, and 12 the anterior inferior cerebellar artery. All the cases were cured through the operations. Fifty patients were followed up for 4 to 8 years, and then 2 recurrent patients were found and other 48 patients had no recurrence. Nine out of fifty patients have facial insensitivity, others facial sense perceptions are right.

CONCLUSIONSThe effect of the operation of retrogasserian combing is sure. This operation was applicable to any case of trigeminal neuralgia, especially to the cases that no vascular compression was found in the operations.

Adult ; Aged ; Decompression, Surgical ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Neurosurgical Procedures ; methods ; Treatment Outcome ; Trigeminal Nerve ; surgery ; Trigeminal Neuralgia ; surgery

OBJECTIVEITS2 of DNA barcoding was used to study genetic polymorphism of Platycodon grandiflorum.

METHODTotal genomic DNA was isolated from P. grandiflorum. PCR was used to amplified the region of internal transcribed spacer 2 (ITS2), and PCR products were sequenced. The sequences of ITS2 were analyzed and compared by Clustal. The intraspecies genetic distance was calculated based on Kimura 2-parameter model by using MEGA 5.05. The ITS2 sequence of Codonopsis pilosula was used as the outreach value for plants of the genus, and the phylogenic tree used constructed by Neighbor-Joining (NJ) method.

RESULTThe K2-P's genetic distance of all samples were ranged from 0 to 0.930. The K2-P's genetic distance of samples at the same area were ranged from 0 to 0.178. The K2-P's genetic distance of samples at different areas were ranged from 0.735 to 0.930. The analytical result showed that the degree of genetic variation were heavy in intraspecies of P. grandiflorum and significantly correlated with geographical location.

CONCLUSIONThe DNA barcoding of ITS2 can applied to study the intraspecific genetic diversity, it provides a reference for further development of DNA barcoding technology applications.

China ; DNA Barcoding, Taxonomic ; DNA, Plant ; genetics ; DNA, Ribosomal Spacer ; genetics ; Molecular Sequence Data ; Phylogeny ; Platycodon ; classification ; genetics ; Polymorphism, Genetic

OBJECTIVETo investigate whether or not lipopolysaccharide (LPS) and ovalbumin (OA) prime rat peripheral leukocytes, the effect of sensitization on priming and the role of phospholipase D in priming.

METHODSThe peripheral leukocytes were separated and purified from sensitized or unsensitized rats. LPS or OA was used as a priming agent and formylmethionylphenylalanine (fMLP) as an activating agent. Degradation of leukocyte was determined by measurement of elastase release and myeloperoxidase (MPO) activity. Phospholipase D (PLD) activity was assayed by the generation of choline,which was measured by choline-oxidase-catalyzed formation of H(2)O(2) and Trinder reaction.

RESULTCompared with cells treated by fMLP alone,leukocytes from unsensitized rat challenged with fMLP after incubated with LPS released more elastase and MPO (P<0.05). But there was no significant difference between leukocytes challenged with fMLP after incubated with OA and fMLP treated alone. In sensitized rat,there was no difference between leukocytes challenged with fMLP after incubated with LPS and fMLP treated alone. But leukocytes challenged with fMLP after incubated with OA released significantly more elastase and MPO than fMLP treated alone (P<0.05). A significant correlation was obtained between the release of elastase and PLD activity (r(s)=0.51,P<0.01), and also between the release of MPO and PLD activity (r(s)=0.73,P<0.01) in unsensitized rat. In sensitized rat, it was 0.48 (P<0.01) and 0.37 (P<0.05) respectively.

CONCLUSION(1) LPS primes peripheral leukocytes from unsensitized rats; (2) OA primes peripheral leukocytes from actively sensitized rats; (3) PLD plays a role in priming of rat peripheral leukocytes.

Animals ; Leukocyte Elastase ; secretion ; Leukocytes ; drug effects ; enzymology ; Lipopolysaccharides ; pharmacology ; Male ; N-Formylmethionine Leucyl-Phenylalanine ; pharmacology ; Ovalbumin ; immunology ; Peroxidase ; blood ; Phospholipase D ; physiology ; Rats ; Rats, Sprague-Dawley

Objective To construct the RNA interference (RNAi) vector targeting the CD40 gene in rats.Methods Effective target sequences that target at CD40 gene were designed,then the oligonucleotide sequences after annealing of the complementary strands were synthetized,the DNA fragments were connected to the GV118 vectors by double digestion with HpaI and XhoI,and the lentiviral vectors which expressed short hairpin RNA were constructed.The lentiviral vectors were identified by PCR and DNA sequencing.Western blot was employed to sort the best interfering targets exogenously,and lentiviral packaging as well as assaying viral titer were also accomplished.Results CD40 siRNA was successfully inserted into the lentiviral vectors and the lentiviral vector was packaged into 293T cells.The determination of virus titer was 2.0 × 1012 TU/L.Conclusions The RNA interference lentiviral vector targeting the CD40 gene in rats was constructed successfully,which will provide the foundation for further exploring the pathogenesis of viral myocarditis and novel therapies in future.

Objective To investigate the effect of CD40siRNA on the pathologic changes and T helper lymphocyte (Th)-17 cells of myocardium and IL-17,IL-23 of serum in rats with experimental autoimmune myocarditis (EAM).Methods Forty 6-8 week old healthy male Lewis rats with body weight of 185-210 g were divided into EAM group,CD40siRNA group,siRNA group,and normal control group randomly,with 10 rats in each group.The rats in EAM group,CD40siRNA group and siRNA group were induced by immunization with cardiac C protein and completed Freund adjuvant in double foot pads.The rats in normal control group were injected with PBS buffer in double foot pads.On the 8th day after immunization,the rats in CD40siRNA group were injected with CD40 siRNA expression vector,and the rats in siRNA group were injected with siRNA expression vector.The rats were sacrificed on day 21 after inoculation.The histopathologic changes were observed by light microscope and the myocardial histopathology scores were calculated.The expression of RORC mRNA of myocardium was detected by real-time quantitative polymerase chain reaction (RTPCR).Enzyme linked immunoabsorption assay was used to determine the serum level of IL-17 and IL-23.Results Compared with EAM group,the myocardial histopathology score(2.34 ±0.60 vs 3.40 ±0.35,P ＜0.05),the expression ofRORC mRNA(2.13 ±0.28 vs 2.93 ±0.36,P ＜0.05) and the serum level of IL-17 (114.38 ± 8.29 vs 148.70 ± 5.04,P ＜ 0.05) and IL-23 (107.00 ± 7.69 vs 136.98 ± 23.16,P ＜ 0.05) were significantly lower in CD40 siRNA group.Conclusions It is suggested that CD40 siRNA expression vector might reduce myocardial injury by inhibiting Th-17 activation and down-regulating the expression of IL-17 and IL-23.

Objective To evaluate the efficacy of 131I treatment for bone metastases from DTC and analyze the survival rates after 131I treatment and prognostic factors. Methods One hundred and six DTC patients with bone metastases treated by 131I during January 1991 and January 2009 were retrospectively analyzed. Treatment efficacy was assessed based on serum Tg change, bone pain palliation and changes on medical imaging. Univariate analysis was performed for defining the factors affecting 131I treatment efficacy. Survival curves were estimated using the life table method. Survival analysis was performed using Kaplan-Meier method. Results Serum Tg decreased dramatically in 37/106 (34.9%) patients treated with131I. Thirty-nine of 61 patients (63.9%) with bone pain had pain relief. Age, tumor subtype and presence of non-osseous distant metastases were significant factors affecting 131I treatment efficacy based on serum Tg change (χ2=6.443, 11.455, 6.756, all P<0.05), but had no significant effects on bone pain palliation (χ2=0, 0, 0.060, all P>0.05). There were no imaging changes of bone metastases in 77.4% of patients after 131I treatment. The overall 5-year and 10-year survival rates from initial diagnosis of bone metastases was 86.47% and 57.90%, respectively. Univariate analysis showed that number of metastases, presence of non-osseous distant metastases and pre-131I treatment surgery were significant factors for survival (Log-rank values were 4.05, 5.98, 4.22, all P<0.05). Conclusions 131I treatment for bone metastases from DTC is effective for lowering serum Tg and palliation of bone pain. Single metastasis, absence of non-osseous distant metastases and pre-131I therapy surgery are favorable predictors of prognosis.

OBJECTIVETo report the serious complications of microvascular decompression and (or) selective neurectomy in cerebellopontine angle.

METHODSTo review 322 cases of microvascular decompression and (or) facial nerve splitting, selective neurectomy for hemifacial spasm, trigeminal neuralgia and glossopharyngeal neuralgia respectively. Among 322 cases, hemifacial spasm 164, which 96 underwent facial nerve splitting, 68 underwent microvascular decompression and facial nerve splitting. The selective neurectomy and neurectomy were performed in 128 cases of trigeminal neuralgia and 30 cases for glossopharyngeal neuralgia respectively.

RESULTSOf the serious complications 2 died (0.6%), The mild to severe hearing impairment occurred in 21 cases (6.5%), in which 3 cases presented total hearing loss (0. 9%); Postoperative cerebellospinal leakage in 20 (6. 2%).

CONCLUSIONSIt was demonstrated that, even though the microvascular decompression and /or neurectomy was a safe procedure for posterior neuropathy, mortal and serious complications might occur. It was essential to pay great attention to the entire decompression procedure in avoiding cerebellar injury. Using of intraoperative ABR and avoiding of impairment of the acoustic nerve might contribute to the prevention of the postoperative hearing loss.

Adult ; Aged ; Cerebellopontine Angle ; surgery ; Decompression, Surgical ; adverse effects ; Female ; Hearing Loss ; etiology ; Humans ; Male ; Middle Aged ; Neurosurgical Procedures ; adverse effects ; Postoperative Complications

BACKGROUNDIn the past 5 years, new treatment materials and techniques offering a different concept in endovascular treatment have been described for cerebral arteriovenous malformations (CAVMs). The aim of this study was to assess the endovascular treatment of CAVMs by using a liquid embolic material, Onyx (Micro Therapeutics. Inc., Irvine, CA, USA).

METHODSFrom September 2003 to September 2004, Onyx was used to treat 22 patients with CAVMs. Ten AVMS were located in functional areas, 8 in deep cerebral areas, and 4 in the cerebellar hemisphere. The size of CAVMs was about 3 cm in diameter in 5 patients, 3-6 cm in 11, and more than 6 cm in 6.

RESULTSIn the 22 patients, Onyx embolization was successful. Nidus occlusion was complete in 3 patients, > 90% in 8, > 80% and < 90% in 6, and > 50% and < 80% in 5. Complications included transient neurological deficits in 2 patients, and adherence of microcatheter to the site of injection in 2.

CONCLUSIONSBeing non-adhesiveness, Onyx is a safe and satisfactory embolic material in the treatment of CAVMs. But its long-term efficacy awaits further follow-up.

Adolescent ; Adult ; Aged ; Child ; Dimethyl Sulfoxide ; administration & dosage ; Embolization, Therapeutic ; adverse effects ; methods ; Female ; Humans ; Intracranial Arteriovenous Malformations ; therapy ; Male ; Middle Aged