A comparison of the volatile compounds in Rhizomes Curcumae (Ezhu) and Radix Curcumae (Yujin) was undertaken using gas chromatography-mass spectrometry (GC-MS).Ultrasonic extraction and GC-MS methods were developed for the simultaneous determination of five sesquiterpenes,namely,α-pinene,β-elemene,curcumol,germacrone and curdione,in Ezhu and Yunjin.Good linearity (r＞0.999) and high inter-day precision were observed over the investigated concentration ranges.The validated method was successfully used for the simultaneous determination of five sesquiterpenes in Ezhu and Yujin.The quantitative method can be effectively used to evaluate and monitor the quality of Chinese curcuma in clinical use.

A comparison of the volatile compounds in Rhizomes Curcumae （Ezhu） and Radix Curcumae （Yujin） was undertaken using gas chromatography mass spectrometi-y （GC-MS）. Ultrasonic extraction and GC-MS methods were developed for the simultaneous determination of five sesquiterpenes, namely, α-pinene, β-elemene, curcumol, germacrone and curdione, in Ezhu and Yunjin. Good linearity （r〉0.999） and high inter-day precision were observed over the investigated concentration ranges. The validated method was successfully used for the simultaneous determination of five sesquiterpenes in Ezhu and Yujin. The quantitative method can be effectively used to evaluate and monitor the quality of Chinese curcuma in clinical use.

In order to study the regularity of shedding virus from infected SPF chickens and the formation of aerosol of H9N2 subtype AIV, SPF chickens were bred in a positive and negative pressure isolator. Aerosol samples were collected by AGI-30 (All Glass Impinger-30) extractor, and simultaneously trachea and cloaca samples were collected by tracheal swabs and cloacal swabs in different periods after challenged with vi- ruses. The above-mentioned samples were detected by HI, Dot-ELISA and RT-PCR methods. The results in- dicated that aerosols were isolated from the 4 days to the 43 days after inoculation. It was proved that H9N2 subtype AIV could copy themselves in respiratory passage and cloaca, and then could formation of aerosols. AIV H9N2 subtype could be isolated from cloacal and tracheal swabs 3 days after inoculation and lasted for 45 days, viruses were detected from all infected SPF chickens on 7 days.

According to the Chinese Pharmacopoeia 2015, only processed Aconitum tubers can be clinically applied, and the effect of processing is unclear. This research aimed to explore the effect of processing on cardiac efficacy of alkaloids in Aconitum tubers. First, the chemical ingredients in unprocessed and processed Aconitum tubers were identified and compared by using high performance liquid chromatography time-of-flight mass spectrometry (HPLC-TOF/MS) and multivariate pattern recognition methods. Then the representative alkaloids in Aconitum tubers, aconitine, benzoylaconine, and aconine, which belong to diester-diterpenoid alkaloids, monoester-diterpenoid alkaloids, and amine-diterpenoid alkaloids, respectively, were selected for further validation of attenuated mechanism. Subsequent pharmacological experiments with aconitine, benzoylaconine, and aconine in SD rats were used to validate the effect of processing on cardiac functions. After processing the Aconitum tubers, it was found that the contents of diester-diterpenoid alkaloids were reduced, and those of monoester-diterpenoid alkaloids and amine-diterpenoid alkaloids were increased, suggesting that diesterditerpenoid alkaloids were transformed into monoester-diterpenoid alkaloids and amine-diterpenoid alkaloids. Through further decocting the aconitine in boiling water, it was confirmed that the three alkaloids could be progressively transformed. Pharmacological experiments with aconitine, benzoylaconine, and aconine in SD rats showed that aconitine at a dose of 0.01 mg/kg and aconine at a dose of 10 mg/kg enhanced the cardiac function, while benzoylaconine at a dose of 2 mg/kg weakened the cardiac function. The effect of processing is attributed to the transformation of the most toxic diester-diterpenoid alkaloids into less toxic monoesterditerpenoid alkaloids and amine-diterpenoid alkaloids.

OBJECTIVE: To develop a multiplex allele-specific PCR (AS-PCR) assay with three-color fluorescence labeling for mitochondrial DNA (mtDNA) SNP typing. METHODS: Based on the principle of AS-PCR, the primer sets were designed for 20 SNP located on the coding region of mtDNA and divided into 2 groups labeled with FAM and HEX fluorescence, respectively. A primer set included two forward (reverse) allelic specific primers with different sizes and a generic reverse (forward) primer. Blood samples from 200 unrelated individuals were analyzed by AS-PCR and capillary electrophoresis. Three random samples at least for each SNP site were examined and verified by direct sequencing. The haplotype frequency was investigated. RESULTS: Distinct electropherograms of 200 blood samples were obtained successfully. The typing results of direct sequencing were identical to those obtained from AS-PCR. The minimum detectable DNA concentration was 0.2 pg under the system of 10 microL. The sensitivity of the DNA concentrations ranged from 0.5 to 5 pg. The 200 individuals were assigned into 15 haplotype, and the haplotype diversity was 0.906 0. CONCLUSION AS-PCR is a simple, rapid and efficient method for mtDNA SNP typing, and can be applied to forensic practice.
Alleles ; DNA ; DNA Primers ; DNA, Mitochondrial/analysis* ; Electrophoresis, Capillary ; Haplotypes ; Humans ; Mitochondria ; Polymerase Chain Reaction/methods* ; Polymorphism, Single Nucleotide ; Sequence Analysis, DNA

Objective-To assess the clinical value of the computer-assisted three-dimensional reconstruction technique design and evaluate the climcal experience of manufacture artificial bone precision to repair the mandibular defect.Methods· From 2001 to 2016,163 computer-assisted reconstruction surgeries had been performed in Craniofacial Department,Ninth People's Hospital,Shanghai Jiao Tong University School of Medicine.During six months followup,the measurement data was conducted and compared with three-dimensional CT result.Random measurement of the three key anatomical points pre-and post-operative carried out with statistical error was used to evaluate the accuracy of computer-assisted three-dimensional reconstruction in mandibular defects repairation and to investigate the clinical application value of the operation time and postoperative complication rate.Results· From July 2001 to July 2016,a total of 163 patients underwent computer-assisted three-dimensional reconstruction of artificial bone repair for mandibular defects;149 patients met the statistical criteria in which preoperative design and postoperative actual effect's average distance error (1.27±0.15) mm,operation time (2.5±1.2) h.Conclusion· Threedimensional design of artificial bone to repair the mandibular defect is a valuable technology,by relying on quantitative design and preoperative simulation to simplify the difficulty and improve the accuracy of surgery.The patients showed high satisfaction rate with low surgical complications and long-term efficacy.

Mast cells participate in allergies and inflammation by secreting a variety of pro-inflammatory mediators. Curcumin, the active component of turmeric, is a polyphenolic phytochemical with anti-tumor, anti-inflammatory, anti-oxidative, and anti-allergic properties. The effects of curcumin on compound 48/80-induced mast cell activation and passive cutaneous anaphylactoid reactions are unknown. In this report, we investigated the influences of curcumin on the passive cutaneous anaphylactoid response in vivo and compound 48/80-induced mast cell activation in vitro. The mechanism of action was examined by calcium uptake measurements and cAMP assays in mast cells. Curcumin significantly attenuated the mast cell-mediated passive cutaneous anaphylactoid reaction in an animal model. In agreement with this in vivo activity, curcumin suppressed compound 48/80-induced rat peritoneal mast cell (RPMC) degranulation and histamine release from RPMCs. Moreover, compound 48/80-elicited calcium uptake into RPMCs was reduced in a dose-dependent manner by curcumin. Furthermore, curcumin increased the level of intracellular cAMP and significantly inhibited the compound 48/80-induced reduction of cAMP in RPMCs. These results corroborate the finding that curcumin may have anti-allergic activity.
Animals ; Calcium ; Curcuma ; Curcumin ; Histamine ; Histamine Release ; Hypersensitivity ; Inflammation ; Mast Cells ; Models, Animal ; Rats

The bear bile has been used as a traditional drug medicine and has been known to have anti-tumor, anti-inflammatory and anti-oxidant effects. The purpose of this study is to investigate the inhibitory effect of bear bile on compound 48/80-induced mast cell activation in vitro and anti-dinitrophenyl (DNP) IgE-mediated vascular permeability in vivo. For this, the effects of bear bile on the degranulation, histamine release, calcium influx and the change of the intracellular cAMP levels of rat peritoneal mast cells (RPMCs) and the influences of the oral treatment of bear bile on IgE-mediated cutaneous vascular permeability were studied. the results were as follows; the compound 48/80-induced degranulation, histamine release and calcium influx of RPMCs were inhibited by pretreatment with bear bile, the cAMP levels of RPMCs were increased by pretreatment with bear bile, and bear bile inhibited anti-DNP IgE-mediated cutaneous vascular permeability. From the above results, it is suggested that bear bile contains some substances which inhibit anti-DNP IgE-mediated vascular permeability and mast cell activation. Bear bile potentially may serve as an effective therapeutic agent for allergic diseases.
Animals ; Antioxidants ; Bile ; Calcium ; Capillary Permeability ; Histamine Release ; Immunoglobulin E ; Mast Cells ; Rats ; Ursidae

Midbrain dopamine (DA) neurons play an essential role in modulating motor control. Defects in central DA neurons affect a wide range of neurological disorders including Parkinson's disease (PD). The greatest motivation in the field has been the potential use of DA neurons for cell transplantation therapy in Parkinsonian patients. Recent studies indicated that BMSCs could differentiate into DA neurons in vitro as neural stem cells (NSC) and embryonic stem cells (ESC) could. However, there are no direct evidences about functional DA neurons derived from BMSCs. According to the protocols which had been applicated in inducing neuronal stem cells and embryonic stem cells differentiate into DA neurons in vitro, the present study provides a protocol by using 50 micromol/L brain derived neurotrophy factor (BDNF), 10 micromol/L forskolin (FSK) and 10 micromol/L dopamine (DA) to induce BMSCs differentiate into DA neurons. After 2 weeks of differentiation, the cells expressed the character of neurons in ultrastructure. RT-PCR discovered mRNA of NSE (neuron specific enolase), Nurr1, Ptx3, Lmx1b and Tyrosine hydroxylase (TH) were positive. Immunocytochemistry staining indicated the ratio of TH-positive neural cells was significantly increased after induced 2 weeks (24.80 +/- 3.36) % compared to that of induction of 3 days (3.77 +/- 1.77) %. And the DA release was also different between differentiated and undifferentiated cells detected by high performance liquid chromatography (HPLC). That is to say BDNF and FSK and DA can induce BMSCs differentiate into DA neurons in vitro, and the transdifferentiated cells express mature neurons characters. BMSCs might be a suitable and available source for the in vitro derivation of DA neurons and cell transplantation therapy in some central neural system diseases such as PD.
Adult ; Aged ; Bone Marrow Cells ; cytology ; metabolism ; ultrastructure ; Brain-Derived Neurotrophic Factor ; pharmacology ; Cell Transdifferentiation ; drug effects ; genetics ; physiology ; Cells, Cultured ; Chromatography, High Pressure Liquid ; Colforsin ; pharmacology ; Dopamine ; metabolism ; pharmacology ; Female ; Humans ; Immunohistochemistry ; Male ; Mesenchymal Stromal Cells ; cytology ; metabolism ; ultrastructure ; Microscopy, Electron, Transmission ; Middle Aged ; Neurons ; cytology ; metabolism ; ultrastructure ; Phosphopyruvate Hydratase ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Tyrosine 3-Monooxygenase ; genetics ; metabolism ; Young Adult

A pairs of primers were designed according to the fusion protein(F)gene sequences of canine distemper virus(CDV)in GenBank.A 369 bp fragment aimed signal peptide fragment of F gene was amplified.The PCR products from viscera samples,blood,urine of fur animals including foxes,minks and raccoon dogs,which collected in the years 2005-2007,were cloned to pMD18-T Vector and sequenced.We obtained 13 positive signal peptide fragments from wild-type strains.The results indicated there was obviously genetic diversity between the wild-type strains and CDV3 and other vaccine strains.The homology with CDV3 is 80.7%-83.2%in nucleotide,and 64.8%-71.3%in amino acid.The analysis for the hydrophobic regions indicated the function of signal peptide fragment may be changed.This study can offer aca- demic data to research of CDV genetic variation and epidemiology.