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

OBJECTIVETo evaluate the reliability, validity, and responsiveness of traditional Chinese medicine (TCM) clinical outcomes rating scale for heart failure (HF) based on patients' report.

METHODSTCM clinical outcomes rating scale for HF (TCM-HF-PRO) were evaluated based on 340 HF patients' report from multiple centers. The completion of the investigation was recorded. Cronbach's α coefficient and split-half reliability were used for reliability analysis, and factor analysis was used to assess the construct validity of the rating scale. Pearson correlation analysis was then used for criterion validity analysis. Discriminant analysis was used to assess the responsiveness of the scale. All 340 HF patients having complete TCM-HF-PRO data were assigned to the treatment group and the control group by central randomization. The total TCM-HF-PRO scores of the two groups were compared using paired t-test to reflect the longitude responsiveness of the scale before treatment and at week 2 after treatment.

RESULTS(1) The recycling rate of the scale was 100.0%. One of them was not filled completely, which was rejected thereby. So the completion rate was 99.7%. The completion time for TCM-HF-PRO scale ranged 15 to 25 min. (2) The Cronbach's α coefficient of rating scale was 0.903, split-half reliability was 0.844 and 0.849. (3) Confirmatory factor analysis showed that 7 factors and items formed according to maximum load factor basically coincided with the construct of the rating scale, 7 factors accumulated contribution rate was 43.8%. TCM clinical outcomes rating scale for HF based on patients' report was relatively better correlated with the Minnesota living with HF questionnaire (r = 0.726, P < 0.01). (4) Discriminant analysis showed that the rating scale correctly classified more than 78.8% of case studies having confirmed initial differential diagnosis by experts. The total scale of the rating scale decreased more in the two group after treatment, with significant difference as compared with before treatment (P < 0.01.

CONCLUSIONTCM clinical outcomes rating scale for HF based on patients' report had good reliability, validity and responsiveness, hence it could be used to assess clinical efficacy for HF patients.

Diagnosis, Differential ; Discriminant Analysis ; Factor Analysis, Statistical ; Heart Failure ; diagnosis ; Humans ; Medicine, Chinese Traditional ; methods ; standards ; Reproducibility of Results ; Surveys and Questionnaires

Objective To explore the CT manifestations and study the value in diagnosing type Ⅲb aortic dissection.Methods CT scanning was performed in 16 cases with type-Ⅲb aortic dissection.The imaging findings were analyzed retrospectively.Results All the cases survived beyond 5 years,12 cases were showed crescent-shaped valve,the false lumen in left was obtained in 15 cases,thrombosis in 9 cases were demonstrated in false lumen and 6 cases showed leakage complications.Conclusion The rate of five years survival is high in cases with type-Ⅲb aortic dissection.CT feature of type-Ⅲb aortic dissection is the crescent-shaped valve with the false lumen in the left.

Background Researches demonstrated that corneal dystrophy is associated with the mutation of transforming growth factor beta induced gene(TGFBI)located at chromosome 5q31 domine.Recent study showed that the gene mutation location is in R124H of TGFBI gene. Objective This study was to identify the mutation characteristics of TGFBI gene in a Chinese family with Avellino corneal dystrophy. Methods This Chinese family with Avellino corneal dystrophy were determined and surveyed in Peking University Third Hospital.Periphery blood from 8 patients with Avellino corneal dystrophy and 2 unaffected subjects were collected from a Chinese family with corneal dystrophy for the extraction of DNA.Exons 4,11,12 of the TGFBI gene were amplified by polymerase chain reaction(PCR),and the amplified products were sequenced directly and compared the gene sequence with that of TGFBI in GenBank.Written informed consent was obtained from each Subject prior to any medieal process. Results This family included 27 members of consecutive 4 generation.The hereditary pattern W88 in accordance with the autosomal dominant inheritance.Directly sequencing of 8 affected members revealed a G tO A transition at codon 124 (CGC to CAC),producing R124H mutation of TGFBI gene.Two synonymous single nucleotide polymorphism(SNP)of TGFBI gene occurred in the family.including a C to T transition at eodon 472(CTC to CTT)in 8 members,and a T to C transition at codon 540(TTT>TTC)in 9 members,which wag unrelated with disease. Conclusion R124H mutation of the TGFBI gene is found in this Chinese family with Avellino corneal dystrophy.

Objective To explore the expression and its implication of angiogenesis and invasiveness related factor in primary and recurrent glioma. Methods Expressions of vascular endothelial growth factor (VEGF), matrix metalloproteinase-2 (MMP-2) and MMP-9 were detected by immunohistochemical technique. The morphological characteristics of ultrastructure of glioma were observed by transmission electron microscope (TEM). Results The expressions of VEGF, MMP-2 and MMP-9 varied in different grades of primary glioma. With the elevation of the malignant degree of the primary glioma, positive staining rates of VEGF, MMP-2 and MMP-9 increased significantly. The expression of VEGF correlated with both MMP-2 and MMP-9 expression. Compared with the primary glioma, the immunoreactivities of VEGF, MMP-2 and MMP-9 in recurrent glioma increased, especially in those with more severe malignancy. Under transmission electron microscope, endothelial cells markedly proliferated and protruded from the deficiency of basemembrane, concomitantly with edema of the extracapillary gap, plasma extravasation as well as some small worm-eaten caverns in the basemembrane.Conclusion VEGF, MMP-2 and MMP-9 play important roles in glioma angiogenesis and invasiveness.Inhibition of their expressions may be a useful therapy to glioma.

The presence of a large palatal or maxillary defect after partial or total maxillectomy for tumor, trauma or congenital deformation poses a challenge to prosthodontists, particularly when the use of an implant cannot be considered. This case report described the use of an air valve in a hollow silicone obturator to manufacture an inflatable obturator that could be extended further into undercut area to retain itself. The inflatable obturator exhibited adequate retention, stability and border sealing, thereby improving the masticatory,pronunciation and swallowing functions of patients. It may be a suitable alternative treatment option to an implant-retained obturator.
Dental Impression Technique ; Dental Prosthesis Design ; Dental Prosthesis Retention ; Denture Retention ; Denture, Complete, Upper ; Fibrous Dysplasia of Bone ; rehabilitation ; surgery ; Humans ; Male ; Maxilla ; surgery ; Orthognathic Surgical Procedures ; instrumentation ; methods ; rehabilitation ; Palatal Obturators ; Silicones ; Treatment Outcome ; Young Adult

OBJECTIVETo investigate the suppressive effect and its frequency selection of dopamine on the cochlear auditory afferent nerve activity. To offer an important step in understanding the modulation of dopamine in the inner cell synaptic complex.

METHODSForty guinea pigs were randomly divided into four groups and the whole intracochlear perfusions were performed: (1) perfused with artificial perilymph solutions; (2) perfused with artificial perilymph solutions containing 10 mmol/L dopamine; (3) perfused with artificial perilymph solutions containing 30 mmol/L dopamine; (4) perfused with artificial perilymph solutions containing 50 mmol/L dopamine. Compound action potential (CAP) evoked by different frequencies (250 Hz, 500 Hz, 1000 Hz, 2000 Hz, 4000 Hz, 8000 Hz, 16 000 Hz) and cochlear microphonics (CM) evoked by 4000 Hz tone burst were recorded from the round window of guinea pigs before perfusion and 1 hours, 2 hours after perfusions.

RESULTSThere was no significant difference in CAP threshold before and after perfusion in the artificial perilymph solutions group (P > 0.05). An increase of CAP threshold of most detecting frequencies were observed in the three dopamine-perfused groups (P < 0.05). The inhibition effect intended with the increasing of the concentration of dopamine in the perfusion solution. There was significant difference in CAP threshold shift between different frequencies, especially in the group perfused with 30 mmol/L dopamine. The maximal threshold shift was found at 4000 Hz and 8000 Hz. No significant changes of CM amplitude and non-linearity of input-output function were observed before and after perfusion with artificial perilymph solutions and dopamine.

CONCLUSIONSDopamine can inhibit the cochlear auditory afferent nerve, but there is no obvious influence on outer hair cell produced by dopamine. The inhibition effect has frequency selection, the suppression on high frequencies is more stronger than low frequencies.

Animals ; Cochlear Nerve ; drug effects ; physiology ; Dopamine ; pharmacology ; Evoked Potentials, Auditory ; drug effects ; Female ; Guinea Pigs ; Male

Objective To study the clinical application anatomy of interventricular foramen and offer a base for operation.Methods Interventricular foramens were observed in 15 adult cadaveric brainThirty- two patients of obstructive hydrocephalus were operated to observe the structure of interventricular foramen un- der neuroendoscope.Results Interventricular foramen was a poriform structure which consists of fornixan- terior pole of thalamencephalon and choroid plexus and was a oval shape in most of themThe plane of the fo- ramen was a included angle with the median sagittal planeThe septal veinthalamostriate veinthalamen- cephal and even the floor of third ventricle could be observed clearly in endoscope.At the same timewe found the foramen had a significant change in obstructive hydrocephalus.Conclusion The interventricular foramen has a simple relatively structure but a variation on size and shape especially in obstructive hydroceph- alusA clearly comprehension of it's structure and adjacent is a base to microsurgery and endoscopic surgery on the foramen.

Fractional Flow Reserve, Myocardial ; Humans ; Male ; Middle Aged ; Tomography, X-Ray Computed ; methods

BACKGROUNDMutations in PAX6 gene have been shown to be the genetic cause of aniridia, which is a severe panocular eye disease characterised by iris hypoplasia. However, there is no study to do genetic analysis of aniridia, although there are several case reports in China. Here, we describe a mutation analysis of PAX6 in a large Chinese family with aniridia.

METHODSGenomic DNA from venous blood samples was prepared. Haplotype analysis was performed with two genetic markers (D11S904 and D11S935). Fourteen exons of the PAX6 gene were amplified from genomic DNA. Polymerase chain reaction (PCR) products of each exon were analysed by single strand conformational polymorphism (SSCP). The PCR products having an abnormal pattern were sequenced to confirm the mutation.

RESULTSSignificant evidence for allele sharing in affected patients was detected suggesting that PAX6 mutation links to aniridia in this family. An extra band corresponding to exon 9 in PAX6 was found by single strand conformational polymorphism analysis in all the aniridia patients in this family, but not detected in the unaffected members. A mutation of C to T was detected by sequencing at the nucleotide 1080 that converts the Arg codon (CGA) to the termination codon (TGA).

CONCLUSIONSAniridia is caused by a nonsense mutation of PAX6 gene in the large Chinese kindred. Genetic test is important to prevent the transmission of aniridia to their offsprings in the kindred by prenatal diagnosis.

Aniridia ; genetics ; Eye Proteins ; genetics ; Female ; Homeodomain Proteins ; genetics ; Humans ; Male ; Mutation ; PAX6 Transcription Factor ; Paired Box Transcription Factors ; Pedigree ; Repressor Proteins ; genetics