Hyoscyamine 6 beta-hydroxylase (H6H) is the last rate-limiting enzyme directly catalyzing the formation of scopolamine in tropane alkaloids (TAs) biosynthesis pathway. It is the primary target gene in the genetic modification of TAs metabolic pathway. Full-length cDNA and gDNA sequences of a novel H6H gene were cloned from Datura arborea (DaH6H, GenBank accession numbers for cDNA and gDNA are KR006981 and KR006983, respectively). Nucleotide sequence analysis reveals an open reading frame of 1375 bp encoding 347 amino acids in the cDNA of DaH6H, while the gDNA of DaH6H contains four exons and three introns, with the highest similarity to the gDNA of H6H from D. stramonium. DaH6H also exhibited the most identity of 90.5% with DsH6H in amino acids and harbored conserved 2-oxoglutarate binding motif and two iron binding motifs. The expression level of DaH6H was highest in the mature leaf, followed by the secondary root, and with no expression in the primary root based on qPCR analysis. Its expression was inhibited by MeJA. DaH6H was expressed in E. coli and a 39 kD recombinant protein was detected in SDS-PAGE. Comparison of the contents of scopolamine and hyoscyamine in various TAs-producing plants revealed that D. arborea was one of the rare scopolamine predominant plants. Cloning of DaH6H gene will allow more research in the molecular regulatory mechanism of TAs biosynthesis in distinct plants and provide a new candidate gene for scopolamine metabolic engineering.

Eurycoma longifolia,also known as Tongkat Ali in Malaysia,as one of traditional herbal medicines,is used for centuries in South-East Asia.With the discovery of anticancer and anti-HIV properties,this herbal medicine has attracted great attention recently.In this review,the following information on E.longifolia,including chemistry,bioactivities,pharmacokinetics,clinical studies,and side effects and safety,was introduced.Our results,to a certain extent,will provide scientific base for commercial utilization and clearance of the Tongkat Ali products with regard to consumers' safety.

AIM: To observe the curative effect of titanium wiring in treatment of the fractures of Neer Ⅱ type distal clavicle and dislocation of Tossy Ⅲ type acromioclavicular joint. METHODS: From March 2004 to December 2006, 37 cases with Neer Ⅱ type distal clavicle fractures or Tossy Ⅲ type acromioclavicular joint dislocation were treated with titanium wiring in Changzheng Hospital, Second Military Medical University of Chinese PLA. Titanium wiring (produced by Sofamor-Danek Company in USA) had good histocompatibility, intensity of tension and flexibility. The broken coracoclavicular ligament was repaired directly by titanium wiring in all cases. The outcomes were evaluated according to Herscovici's criteria. RESULTS: All the cases were followed up for 6 to 24 months. All the dislocated acromioclavicular joints returned to normal 2 weeks after operation. Excellent and good rate of shoulder function was 100% according to Herscovici standard. X-ray imaging showed all clavicle fractures united, acromioclavicular joint had no redislocation or titanium wiring breaking. The incisions of all cases had no obvious swelling and exudation. CONCLUSION: When Neer Ⅱ type distal clavicular fracture and Tossy Ⅲ type acromioclavicular joint dislocation are treated with titanium wiring, the acromioclavicular joint can exercise earlier and recover quickly.

Objective To observe the abnormality of gray matter volume and density in patients undergoing postoperative delirium (POD).Methods Forty-seven cases of aged patients, 26 males and 21 females, aged 60-75 years, ASA physical status Ⅱ or Ⅲ, were selected.On the third day after operation, cognitive function estimation was performed.The patients were assigned into group POD and group C according to whether POD occurred and brain magnetic resonance imaging (MRI) scanning was implemented.The discrepancy in gray matter volume and density between the two groups were compared using voxel-based morphometry method (VBM).Correlation analysis was performed between the corresponding parameters in the regions where notable differences between the two groups existed and minimum mental state examination (MMSE) score were found.Results Global gray matter volume of group POD was notably lower than that of group C (P<0.01).Cerebrospinal fluid volume of group POD was significantly higher than that of group C (P<0.01).Gray matter volume of bilateral frontal gyrus and right parahippocampus was remarkably reduced in group POD (P<0.001).Gray matter density of bilateral hippocampus and right parahippocampus decreased significantly (P<0.001).Right parahippocampal gray matter volume was positively correlated with MMSE score in POD patients (P<0.05).Conclusion Structural abnormality in frontal regions, hippocampus and parahippocampus may play an important role in pathogenetic and developing process of POD.Gray matter volume in the right parahippocampus may be one of reference index for POD severity.

Objective To investigate the alteration of serotonin-producing gastric enterochromaffin (EC)cell in patients with functional dyspepsia(FD).Methods Fifteen healthy volunteers and 33 patients with FD were enrolled.Proximal gastric mucosal EC cells were countered after immunohisto- chemistry staining.The ultrastructure of EC cell was observed by electromicroscope.Results The EC cells in proximal gastric mucosa in patients with FD were significant higher than that in controls(12.5?2.1 vs 8.3?1.4,t=2.353,P＜0.05),and the staining intensity of EC cell in patients with FD was also stronger than that in controls(3.72?0.42 vs 2.61?0.57,t=2.078,P＜0.05).The more sever the gastric mucosal inflammation was,the more number of EC cells and the stronger staining intensity were.Under the electromicroscopy,more Golgi apparatus,mitochondria and endoplasmic reticulum were found in EC cells.Special secreting particles were also found in cytoplasm.Conclusions EC cells may be involved in the pathogenesis of FD.The number of EC cell is related with the severity of gastric mucosal inflammation.

Background The identification of mesenchymal stem cells(MSCs) have provided exciting prospects for cell-based regeneration after myocardic infraction.However cell therapy have inherent limitations such as low survival rate of transplanted cells and insufficient cell number.It is known that cell-matrix adhesion plays a key role in cell proliferation,differentiation and survival,and chemokine CXCL12 may involved in these prcesses.Transfected mesenchymalstem cells with CXCL12 for local secretion of CXCL12 and then explored CXCL12 triggered adhesion of mesenchymal stem cells to extracellular matrix proteins.Mesenchymal stem cells was transfected with CXCL12.?V and ?3 integrins content was evaluated by Western blot analysis.Cell adhesion to extracellular matrix was examined in vitro and cell prolife-ration after transplantation in vivo.Transfection of CXCL12 resulted increased CXCL12 in situ.Increased CXCL12 induced elevated adhesion to fibronectin in vitro and higher survival in vivo.CXCL12 mediated adhesion and proliferation was established by ?V and ?3 integrin subunits.Chemoattractive mechanisms are involved in adhesion processes of mesenchymal stem cells.Increased CXCL12 leads to enhanced expression of ?V and ?3 integrins,which may augment cell survival,proliferation and differrentiation.

Current occupational health and occupational medicine teaching content focused on the detection of harmful factors,and more belonged to validation experiments.Closed-end management of experimental teaching,experimental report-based performance evaluation methods and single model of teaching methods reduced the students' study interesting,lack of problem-solving abilities.Therefore,it is necessary to optimize the experimental curriculum,adopt an open experimental teaching,build a reasonable experimental evaluation system and promote the diversification of teaching model.

Objective To investigate the mitochondrial energy metabolism in D-galactoseinduced cell ageing model.Methods MRC-5 cells were cultivated for 72 hours in a medium containing 55 mmol/L D-galactose.The analysis of cell proliferation capacity by CCK8 method,β-galactosidase staining and detection of p21 protein expression level were performed for identifying cell senescence.The cell oxidation-reduction state was evaluated by an analysis of cellular ROS levels,SOD activity,MDA content and oxidative damage level of mitochondrial DNA(mtDNA).For purpose of detecting mitochondrial function and its impairment,mitochondrial morphology was observed by electron microscope,mitochondrial quantity was analyzed by flow cytometry,mitochondrial membrane potential(△Ψm) was measured by JC-1 staining,and ATP content was analyzed by HPLC,and mitochondrial oxygen consumption rate was detected by Seahorse cell energy metabolism detection system.Results The decreased MRC-5 cell proliferation,up-expression of p21 protein,increased β-galactosidase activity were observed in D-Gal-treated cells,which indicated the cell premature senescence.When treated with D-Gal,the significantly increased ROS and MDA level,decreased SOD activity and increased oxidized mtDNA proved that the cells kept higher oxidative stress.D-Gal induced-mitochondrial impairment was evidenced by the dimming of mitochondrial cristae and double membrane structure,decrease of transmembrane potential and ATP synthesis,and decrease of its oxygen consumption rate(OCR).Conclusions The 55 mmol/L D-Gal causes an impairment of mitochondrial structure and a decrease of function of energy metabolism,which is associated with cellular senescence induced by D-Gal.

Referring to the rules for agricultural seed testing (GB /T 3543-1995) issued by China, the test of sampling, seed purity, weight per 1 000 seeds, seed moisture, seed viability and germination rate had been studied for screening seed quality test methods of Pesudostellaria heterophylla. The seed quality from different collection areas was measured. The results showed that at least 6.5 g seeds should be sampled and passed through 10-mesh sieve for purity analysis. The weight of 1 000 seeds was determined by using the 500-seed method. The phenotypic observation and size measurement were used for authenticity testing. The seed moisture was determined under the higher temperature (130 ± 2) degrees C for 5 hours. The seeds were dipped into 0.2% TTC sustaining 1 hour at 40 degrees C, then the viability could be determined. The break dormancy seeds were cultured on sand at 10 degrees C. K cluster analysis was applied for the data analysis, the seed quality from different collection areas grading of P. Heterophylla was described as three grades. The seed quality of each grade should reach following requirements: for first grade seeds, germination rate ≥ 86%, 1 000-grain weight ≥ 2.59 g, purity ≥ 87%, moisture ≤ 13.1%; for second grade seeds, germination rate ≥ 70%, 1 000-grain weight ≥ 2.40 g, purity ≥ 77%, moisture ≤ 14.3%; for third grade seeds, germination rate ≥ 41%, 1 000-grain weight ≥ 2.29 g, purity ≥ 76%, moisture ≤ 15.8%. The seed testing methods for quality items of P. heterophylla had been initially established, as well as the primary P. heterophylla seed quality classification standard.
Caryophyllaceae ; chemistry ; growth & development ; Germination ; Quality Control ; Seeds ; chemistry ; growth & development ; Water ; analysis