[Association of fimA genotype to kgp genotype in Porphyromonas gingivalis from infected root canals of primary apical periodontitis].

Objective: To investigate the distribution of fimA and kgp genotypes as well as the common genotype combination of Porphyromonas gingivalis (Pg) in infected root canals of primary apical periodontitis for virulent isolates screening in future. Methods: Thirty-four samples harboring Pg were selected from infected root canals of primary apical periodontitis from patients of the Department of Endodontics, Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine from June 2013 to September 2015. FimA type-specific primers were used to amplify the samples, revealing the distribution of various fimA genotypes. The genotypes of kgp were obtained by using Mse Ⅰ restriction endonuclease. The prevalence of each genotype and common genotype combinations were then calculated. Pearson's chi-squared test was performed to analyze the correlation between genotype combinations and clinical symptoms and major signs of apical periodontitis. In addition, the bioflim architectures between Pg isolates with different fimA and kgp genotype combinations were observed compared using confocal laser scanning microscope. Results: Among the 34 Pg-positive samples, fimA Ⅱ was the most prevalent genotype [47% (16/34)] followed by fimA Ⅰ [26% (9/34)], while fimA Ⅴ was detected in only one sample. The prevalence of kgp Ⅰ [56% (19/34)] was slightly higher than that of kgp Ⅱ [44% (15/34)]. Both fimA Ⅱ+kgp Ⅰ and fimAⅡ+kgp Ⅱ were the most prevalent genotype combinations [24% (8/34) each]. No significant correlation was found between specific genotype combination and such major clinical manifestations as gingival swelling and sinus tract of dental origin (P>0.05). Three Pg isolates with different genotype combinations were acquired. Isolate A (fimAⅠ+kgpⅠ) formed densest biofilm, while the biofilm of isolate C (fimAⅤ+kgp Ⅰ) was much looser. The biofilm feature of isolate B (fimAⅢ+kgp Ⅱ) fell in between A and C. Conclusions: Pg with fimA Ⅱ was most frequently detected in infected root canals of primary apical periodontitis. The prevalence of Pg with kgp Ⅰ was slightly higher than that with kgp Ⅱ, and fimAⅡ+kgp Ⅰ as well as fimA Ⅱ+kgp Ⅱ were the commonest genotype combinations. According to the comparison of Pg biofilms formed by clinical isolates, it might be possible that different genotype combinations may lead to distinct biofilm architectures.

An optimized mini-preparation method to obtain high-quality genomic DNA from mature leaves of sunflower.

In order to investigate the mutation characteristics and to further examine the genetic variation of mutant sunflower (Helianthus annuus) obtained in plants grown from seeds exposed to space conditions, only the mature tissues such as leaf and flower could be used for DNA extraction after mutation characteristics were confirmed. The rich contents of polysaccharides, tannins, secondary metabolites, and polyphenolics made it difficult to isolate high-quality DNA from mature leaves of sunflower according to previous reports. Based on the comparison of the differences in previously reported protocols, a modified method for the extraction of high-quality DNA was developed. Using this protocol, the DNA isolated from sunflower was high in quality and suitable for restriction digestion (EcoRI, HindII, BamHI), random amplified polymorphic DNA study and further molecular research. Therefore, the modified protocol was suitable for investigating the genetic variation of sunflower using mature leaf DNA.

[Stable inheritance of hpt gene in transgenic rice plants mediated by biolistic bombardment].

Japonica rice cv 77170 was transformed with pBI222 carrying hygromycin phosphotransferase gene (hpt) by using biolistic bombardment and fertile transgenic rice plants were obtained. In T1 and most T2 generation the performance of hygromycin resistance was normal. Only in T2 progeny of SG-15 some lines showed decreased resistance. Mendelian inheritance of hygromycin resistance was showed as single dominant locus and proved by Southern blotting analysis in T1 and T2 generation of all transgenic plants. Multiple copies of hpt integrated into rice genome. These copies linked closely and inherited stablly. Msp I digested Southern blotting showed that methylation of hpt existed commonly in transgenic plants and their progeny, therefore uncomplete silencing of hpt in some T2 progeny of SG-15 was not associated with hpt methylation probably.

[The factors of improving rice transformation efficiency].

The factors influencing the rice transformation frequency have been investigated by using 8 indica and japonica rice cultivars during the transformation procedures, such as induction of calli, selection, and regeneration. The following is the results: The immature embryos were pretreated in 4 degrees C for 4 or 7 days before inoculation, and the regeneration frequency of calli would be increased. High osmotic treatment may increase the possibility of transformation, especially for calli of 6 months or older. The transformation frequency was improved when selection pressure was removed during differentiation period. The V-type selection may be better than the normal selection. There is a difference of differentiation ability among the calli of three periods and it is found that the calli of two or three months are the best for transformation. Proline and DMSO can decrease browning rate of calli.

[Improvement of transformation frequency of rice mediated by Agrobacterium].

The factors influencing the frequency of rice transformation mediated by Agrobacterium have been investigated by using 16 commercially important indica and japonica rice cultivars or lines. The main results were as following: For most rice CC medium was the best for both callus initiation and subculture. With supplement of 2.5-5 mg/L ABA the quality of calli can be improved. The concentration of selective agent for Indica rice callus was lower than that for japonica rice callus. Agrobacterium tumefaciens strain EHA105 was more efficient than LBA4404 and AGL1 for rice transformation. The inhibitive effect of cefotaxime to Agrobacterium was better than that of carbenicillin. The partial desiccation treatment after co-cultivation was beneficial to inhibit the growth of Agrobacterium and increase transformation efficiency. A stable and efficient Agrobacterium-mediated transformation system has been established in ten different rice cultivars and fertile transgenic plants have been obtained.

[Differential expression of cardiac immediate early and late response gene in exercise-induced and hypertensive cardiac hypertrophic remodeling].

Our previous studies have shown that the myocardial phenotypes between exercise-induced and hypertensive cardiac hypertrophy are different. However, alteration of cardiac gene expression in the two kinds of hypertrophy is not clear. We therefore studied the expression of immediate early and late responsive gene in Wistar rats subject to a 12-weeks swimming program and spontaneously hypertensive rats (SHR) by Northern blot. The autoradiographs were analyzed by scanning densitometry. When compared with matched sedentary control, swimming rats and SHR increased their heart/body weight by 29% and 61% respectively (P < 0.01). After a series of swimming program, levels of c-fos and atrial natriuretic factor (ANF) mRNA were instantly increased 203% and 214% respectively in the swimming rats. However in 24 h after swimming, ANF mRNA was decreased to the level of control but c-fos mRNA was still higher than than in control. The levels of c-fos and ANF mRNA were increased by 80% and 298% in SHR respectively. The expression of beta-myosin heavy chain (MHC) mRNA was increased by 117% in SHR but decreased by 12% in swimming rats. The expression of alpha-MHC mRNA was decreased by 42% in SHR but unchanged in swimming rats. The abundance alpha/beta-MHC mRNA in swimming rats is 336% higher than in SHR. These results showed differential expression of the immediate early and late responsive gene in exercise-induced and SHR hypertrophied heart remodeling. It is suggested that (1) the differential expression of c-fos gene may differently regulate remodeling of these two cardiac hypertrophies; (2) the differential expression of ANF gene may be a molecular marker for distinguishing the two kinds of cardiac hypertrophy; and (3) the differential expression of alpha/beta-MHC gene ratio may regulate exercise-induced and hypertensive hypertrophied myocardial cell changing to contractile phenotype and synthetic phenotype respectively, at the gene transcription level.

Thin sectional anatomy, three-dimensional reconstruction and visualization of the heart from the Chinese Visible Human.

The research aimed to provide sectional anatomic and three-dimensional (3D) virtual anatomic bases for imaging diagnosis and surgical operation by the use of data from the heart of the first Chinese digitized Visible Human. Data from the series of thin sections of the heart were analyzed and input into an SGI workstation, and 3D reconstruction and virtualization of the heart were performed. Each image of sectional anatomy was clear and the 3D structures of the heart were reconstructed in their entirety. All reconstructed structures can be displayed by multiple structural and color modes, individually or jointly, and can be rotated continuously in any plane. The model of the virtual heart clearly showed fine structures of the heart in random orientation. The dataset of the sectional anatomy provides a fine and integrated morphologic base for imaging diagnosis. The 3D reconstructed images clearly show the internal and entire structures of the heart.

Bilateral fetal grafts for Parkinson's disease: 22 months' results.

Five patients with severe Parkinson's disease underwent bilateral multiple graft implants of nondissociated fetal mesodiencephalic tissues. Graft implantation was performed in China following CT-guided stereotactic placement of a novel delivery system. Follow-up has demonstrated substantially reduced levodopa requirements and clinical improvements of motor, postural functions and reduction of freezing and on-off phenomenon. PET utilizing [18F]-dopa, at 14 months in the first case, suggested graft-induced restoration of dopaminergic transmission in the striatum.

Computerized 3D-reconstructions of the ligaments of the lateral aspect of ankle and subtalar joints.

3D-reconstruction images of the structures of lateral aspect of the ankle and subtalar joints were produced using plastination to make equidistant serial sections of 1.2 mm in thickness. A SGI workstation was employed to reconstruct the structures of the ligaments of the lateral aspect of ankle and subtalar joints in three dimensions. Reconstructed structures were displayed singly, in groups or as a whole, and these were rotated continuously at different velocities in 3D space. Different diameters and angles of the reconstructed structures could be measured easily. Improved results could be achieved with the use of a special sectional anatomical technique, i.e. contours + marching cubes algorithm.

Visualization of the temporal bone of the Chinese Visible Human.

Temporal bone anatomy is very difficult to understand. After dataset acquisition of the first Chinese Visible Human, we processed the two-dimensional images to build a digitized visible model of the temporal bone and explore the role of virtual endoscopy in the inner ear. On a SGI workstation three-dimensional computer reconstructions of the ear were generated from the Chinese visible human dataset, viewing the middle and inner ear imitating the traditional otoscopy. The three-dimensional data of the temporal bone were then converted to STL format, and the temporal bone replica were fabricated with rapid prototyping by laminated object manufacturing. The virtual model of the ear was successfully completed, and the virtual endoscopy improved three-dimensional visualization of the middle and inner ear. Physical replica of the temporal bone were built with paper; the accuracy was +/-0.2 mm. The reconstructed model and the replica of the temporal bone can be used to make preoperative plans in the complicated otoneurosurgical operations, allowing various surgical exercises to be carried out on the three-dimensional stereophysical model. The virtual endoscopy stands as a promising new visualization technique for elucidation of the middle and inner ear and reveals a tremendous potential in both clinical and educational settings, providing morphological data for the image diagnosis and otoneurosurgery.

The pre-styloid compartment of the parapharyngeal space: a three-dimensional digitized model based on the Chinese Visible Human.

To build a digitized visible model of the parapharyngeal space of the Chinese Visible Human and to provide a sectional anatomic basis for radiological and clinical diagnosis of the parapharyngeal space, sectional anatomy data of the parapharyngeal space were selected from the Chinese Visible Human male and female to compare with MR imaging findings in the axial planes. From these data the parapharyngeal space and surrounding structures were segmented. They were then reconstructed in three dimensions on PC. In the axial planes of the sectional anatomy and MR imaging, the shape, content and relations of the parapharyngeal space were clearly displayed and the dominant plane for showing the parapharyngeal space was elicited. The three-dimensional reconstructed images displayed perfectly the anatomic relationships of the parapharyngeal space, parotid, muscles, mandible and vessels. All reconstructed structures can be displayed singly, in groups or as a whole; any diameter or angle of the reconstructed structures can be easily measured. The Chinese Visible Human male and female data set can provide complete and accurate data. The digitized model of the parapharyngeal space and its surroundings offers unique insights into the complex anatomy of the area, providing morphologic data for imaging diagnosis and surgery of the parapharyngeal space.

An optimized mini-preparation method toobtain high-quality genomic DNA from mature leaves of sunflower

In order to investigate the mutation characteristics and to further examine the genetic variation of mutant sunflower (Helianthus annuus) obtained in plants grown from seeds exposed to space conditions, only the mature tissues such as leaf and flower could be used for DNA extraction after mutation characteristics were confirmed. The rich contents of polysaccharides, tannins, secondary metabolites, and polyphenolics made it difficult to isolate high-quality DNA from mature leaves of sunflower according to previous reports. Based on the comparison of the differences in previously reported protocols, a modified method for the extraction of high-quality DNA was developed. Using this protocol, the DNA isolated from sunflower was high in quality and suitable for restriction digestion (EcoRI, HindII, BamHI), random amplified polymorphic DNA study and further molecular research. Therefore, the modified protocol was suitable for investigating the genetic variation of sunflower using mature leaf DNA.