Tropane alkaloids,such as hyoscyamine (or its more stable racemate atropine) and scopolamine,are remembered as the oldest drugs in medicine with wide pharmaceutical applications clinically for their mydriatic,antispasmodic,anticholinergic,analgesic and sedative properties.Presently,the supply of tropane alkaloids are entirely rested on their isolation from plant materials,yet the low contents of tropane alkaloids in plants raise the requirement to breed new varieties of high-yield tropane alkaloids and improving the production of tropane alkaloids in hairy root cultures or plants by biotechnology has be a research focus in the field of secondary metabolism.For the past decade,there have been important progresses on biosynthesis of tropane alkaloids and their molecular biology,some new pathway genes have been identified and many achievements have also been acquired in metabolic engineering of tropane alkaloids.This review summarized the recent advances in above aspects,and the problems and orientations of future research are also discussed and proposed.

Objective: To establish an accelerated solvent extraction(ASE)-HPLC method to determine chrysophanol and aurantio-obtusin in Cassia obtusifolia L.Methods: The optimal extraction conditions were defined by orthogonal tests using ASE.The method was carried out on an ACE Excel C18-PFP column (75 mm×2.1 mm,2.5 μm) with the mobile phase consisting of 0.1% phosphoric acid solution-acetonitrile with gradient elution.The column temperature was 40 ℃,the flow rate was 0.4 ml·min-1, and the detection wavelength was 284 nm. Results: The best process parameters of ASE were as follows:the extraction solvent was methanol, the extraction temperature was 120 ℃ and the static extraction duration was 5 minutes for three cycles.The ASE method needed only 1/9 of the time as the pharmacopoeia method,while the extraction efficiency of the ASE method was higher.The linear ranges of cassia obtusifolia L.and Chrysophanol were at 0.73~58.57 μg·ml-1(r=0.999 7) and 1.09~87.29 μg·ml-1(r=0.999 6).The average recoveries were 102.7%(RSD=0.8%) and 98.2%(RSD=1.5%).Conclusion: The method is simple, rapid and sensitive, which can be used for the rapid determination of aurantio obtusin and chrysophanol in Cassia obtusifolia L.

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.

To meet the requirement of curriculum design about medical imaging field to undergraduate in medical college,the course of medical imaging radiation protection was developed by compiling a new textbook,establishing curriculum planning,implementing and evaluating teaching system.The teaching system of “one aim and two follows” was set up by use of three stages and three cycle feedbacks research and the course construction was promoted and developed by its teaching assessment,summary in time and the innovation of the teaching method.

Bilirubin ; blood ; Breast Feeding ; Evoked Potentials, Auditory, Brain Stem ; Female ; Humans ; Infant, Newborn ; Jaundice, Neonatal ; physiopathology ; therapy ; Male

Objective To explore the effect of reconstructing unilateral cleft lip by changing the arc-shaped incision, combined with the 3D reconstruction of upper lip muscles.Methods Twenty unilateral cleft lip patients were treated by using a new surgical operation, the 3D reconstruction of upper lip muscle, to restore normal anatomy and stress of the mucous membrane, muscle and skin.Operation scar was designed for straight line, located on the philtral ridges of the contour line;phitrum and philtral ridges were rebuilt, and postoperative scar reduced.Results A lot of 20 patients had no local infection, hemorrhage, complex crack, and were stage I incision healing.Followed up for 1-8 months postoperatively, the patient's lip bow line continuity was good, with symmetrical shape and good phitrum and philtral ridges;scar was hidden on the philtral ridges of the contour line, and no obvious upper lip scar contracture found through the follow-up period.Conclusions This improved method is simple in the incision design, and less scar hidden on the philtral ridges of the contour line after operation, which can maximize the recovery of the appearance of nose and upper lip with satisfactory effect.It is a feasible improvement method of repairing unilateral cleft lip.

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.
Cloning, Molecular ; DNA, Complementary ; Datura ; enzymology ; genetics ; Escherichia coli ; Hyoscyamine ; chemistry ; Mixed Function Oxygenases ; genetics ; Plant Leaves ; enzymology ; Plant Roots ; enzymology ; Recombinant Proteins ; genetics ; Scopolamine Hydrobromide ; chemistry

Objective To explore the cause, influencing factor and possible solving method of transient adverse reactions during the operation. Methods Sixteen healthy beagles were undergone virtual PLDD. The changes of vertebral discs and the surrounding tissues were observed by high resolution CT and MRI at different periods after the operation, and the same investigation procedure was carried out after the sacrifice of beagles. Results The beagles of the conventional vaporization group occasionally had limb tic and whining in the operations. Pieces of necrosis and edema could be found in the tissues of intervertebral foramen at the paracentetic side. The histological changes in the negative pressure suction group were less than those in the conventional group. Conclusion The reversible damages of the surrounding tissuses were observed in the conventional group and continuing negative pressure suction during the operations can prevent the damages to the surrounding tissues, all of the changes could be clearly displayed by CT and MRI scan.

Objective To explore the operation opportunity of cholecystectomy for senile patients with acute cholecystitis combined with underlying disease.Methods 82 elderly patients with acute cholecystitis complicated with underlying diseases underwent cholecystectomy were selected,they were divided into two groups according to the different operation time:the early operation group and the delayed operation group,41 cases in each group.The patients in early operation group were given operation in 72h,while the delayed operation group was given operation more than 72h after attack.The operation time,intraoperative blood loss,intraoperative tissue adhesions,intraoperative exudation,hospitalization time,cost of hospitalization and complications of the two groups were observed and com-pared.Results The operation time of the two groups were (45.32 ±1 2.23)min,(50.89 ±1 3.34)min,there was no significant difference (P >0.05),the intraoperative tissue adhesions of the delayed operation group was more than the early operation group,and the intraoperative hemorrhage and exudation in the early operation group were significantly higher than those of the delayed operation group (P <0.05),and the hospital stay and cost of hospitalization were sig-nificantly lower than those of the delayed operation group (P <0.05).1 case in the early operation group occurred pulmonary infection,3 cases with incision infection,the incidence of complications was 9.76%;1 case in the delayed operation group underwent laparotomy operation,2 cases in the delayed operation group had pulmonary infection,inci-sion infection in 4 cases,the incidence rate of complication was 1 4.63%,there was no significant difference between the two groups (P >0.05).Conclusion The optimal operation timing of acute cholecystitis combined with basic dis-eases in the elderly patients underwent cholecystectomy is within 72h.For patients who can not be immediately given operation due to various reasons,delayed cholecystectomy is safe and feasible.

OBJECTIVETo explore the effect of microneedle combined with Lauromacrogol on skin capillary network.

METHODS24 male Leghone (1.5-2.0 kg in weight) were randomly divided into three groups as group A (microneedle combined with Lauromacrogol), B (microneedle combined with physiological saline) , and C(control). The cockscombs were treated. The specimens were taken on the 7th, 14th, 21th , and 28th day postoperatively. HE staining, immunohistochemical staining and special staining were performed for study of the number of capillary and collagen I/III , as well as elastic fibers.

RESULTSThe color of cockscombs in group A became lightening after treatment. The number of capillary decreased as showing by HE staining. The collagen I and III in group B was significantly different from that in group A and C (P < 0.05). Special staining showed proliferation of elastic fibers in group B.

CONCLUSIONSIt indicates that microneedle combined with Lauromacrogol could effectively reduce the capillary in cockscomb without any tissue fibrosis. Microneedle can stimulate the proliferation of elastic fiber, so as to improve the skin ageing process.

Animals ; Capillaries ; anatomy & histology ; Chickens ; Comb and Wattles ; blood supply ; drug effects ; Male ; Needles ; Polyethylene Glycols ; pharmacology ; Punctures ; instrumentation ; methods ; Random Allocation ; Skin Aging