Pulsed field gel electrophoresis(PFGE)is a new type technique of gel electrophoresis which can be used to separate large DNA molecules.It has been widely applied to the karyotype analysis,identification of species groups,genetic orientation and genetic analysis for fungi.This article describes the principle,development and general manipulative procedure of PFGE,and elaborates the application in the molecular research of fungi.

OBJECTIVE:To optimize the extraction technology of Bushen huoxue capsules. METHODS:Central composite de-sign-response surface method was used to optimize the extraction technology with the amount of water and boiling time as main fac-tors using normalized value of the contents of icariin and salvianolic acid B,the yield of dry extract as index. Validation test was al-so conducted. RESULTS:The optimal extraction technology was as follows as 15-fold water,reflux extracting for 75 min,extract-ing for 2 times. The deviation of measured value and predicted value was 0.000 3% in validation test. CONCLUSIONS:The cen-tral composite design-response surface method can be used to optimize the extraction technology of Bushen huoxue capsules with good prediction,and the technology is simple and stable.

Study a method for the detemination of the content of polysaccharides in Gentiana farreri, and analysis of the content of polysaccharides from different producing areas. The results showed that using the anthrone-sulfuric acid method, simple operation, accurate result. Sample was measured at 620 nm absorbance after anthrone-sulfuric acid color, at this wavelength, solution absorption and glucose showed a good linear relationship; The linearity was in the range of 0.01-0.07 g x L(-1) (r = 0.996 7). The recovery rate was 99.41%, with RSD of 2.0%. Considering the experimental conditions, to determine the solid-liquid ratio 1:60, extracting time 50 min, concentration of ethanol 80%. The mass fraction of polysaccharides was the highest to reached 0.743% in G. farreri from Gansu Xiahe. This experiment has laid a good foundation for further study on G. farreri.
Anthracenes ; chemistry ; Chemistry Techniques, Analytical ; methods ; Gentiana ; chemistry ; growth & development ; Geography ; Linear Models ; Polysaccharides ; analysis ; Reproducibility of Results ; Sulfuric Acids ; chemistry ; Time Factors

12E7 Antigen ; Adolescent ; Adult ; Antigens, CD ; metabolism ; Biomarkers, Tumor ; metabolism ; Brain Neoplasms ; secondary ; Cell Adhesion Molecules ; metabolism ; Child ; Child, Preschool ; Diagnosis, Differential ; Extremities ; Female ; Follow-Up Studies ; Humans ; Immunohistochemistry ; In Situ Hybridization, Fluorescence ; Infant ; Ki-67 Antigen ; metabolism ; Male ; Neuroectodermal Tumors, Primitive ; metabolism ; pathology ; Oncogene Proteins, Fusion ; metabolism ; Repressor Proteins ; metabolism ; Sarcoma, Ewing ; metabolism ; pathology ; Sarcoma, Synovial ; diagnosis ; metabolism ; pathology ; surgery ; Soft Tissue Neoplasms ; diagnosis ; metabolism ; pathology ; surgery ; Vimentin ; metabolism ; Young Adult

Objective To establish a method for studying molecular mechanism of Rhubarb inhibiting anti-Yersinia pesti based on DNA microarray.Methods A whole genome DN A microarray containing 4005 annotated genes of Yersiniapesti Was used.The minimal inhibitory concentration(MIC)of Rhubarb to Yersiniapestiwas determined by liquid dilution method.The gene expression profile of Yersinia pesti was performed after the exposure to Rhubarb at a concentration of 10×MIC for 30 minutes.The total RNA extracted and purified from Yersinia pesti Was reversely transfected to cDNA and labeled by Cy3-Cy5 dye.The labeled probes were hybridized to the microarray anti the results were obtained by a laser scanner and the microarray data was confirmed by real-time quantitative RT-PCR.Results The platform of the DNA microarray-based bacteria transcriptional profile was established.A total of 498 genes of Yersinia pesti changed significantly in response to Rhubarb.Among them.358 genes were up-regulated,140 down-reguated.Conclusions The whole genome DNA microarray can be used in the studying of molecular anti-Yersinia pesti mechanism of Rhubarb.

To study the clinical skills training methods and formulate a training objective for pediatric long-year-program students, to let the students, through three years of training program, reach the clinical ability as a chief resident, with better laboratory research skills, teaching awareness and responsibility, and also with good comprehensive quality and excellent English, and finally become clinical talents with great potential and innovation, this report summarized our study results of training two terms of long-year-program pediatric graduates, who fully meeted the training objectives in the department's practice.

Objective To construct the bp26 deletion mutant of Brucella vaccine strain M5-90 (M5-90Δbp26),to compare its immunogenicity with parental strain(M5-90),and to develope a new candidate vaccine strain of Brucella melitensis with reduced virulence that can be used to distinguish vaccinated livestock from infected animals.Methods Mutant vaccine strain of Brucella melitensis was constructed by conventional molecular biology techniques then the genetic stability of mutant M5-90Δbp26 was tested and its conventional bacteriological nature was identified; 1.0 × 109 CFU/2 ml doses of M5-90Δbp26 strain and the parental strain were used to vaccinate 3 sheep; sera were analyzed for reactivity against BP26 by Western blotting and for agglutination activity; to analyze the virulence of mutant and parental strain,mice were injected with 1.0 × 106,6.0 × 106 and 2.0 × 107 CFU/0.2 ml doses of M5-90 and M5-90Δbp26,respectively,and clinical symptoms were monitored and the death of mice was recorded.Results The M5-90Δbp26 was successfully generated and reversion was not observed in 15 generations.The size of PCR products was 629 bp while the parental strain was 1279 bp.The sequence analysis showed a 650 bp missing in M5-90Δbp26.The conventional bacteria identification tests confirmed that the mutant was depth variant strain,including mono-specific antiserum M type transformed into R,and the BK2 phage based splitting assay converted from the positive to the negative.Western blotting showed the purified BP26 protein was recognized by the serum against the parental strain while not by the serum against M5-90Δbp26 strain.Agglutination test showed the level of the serum antibody induced by M5-90Δbp26 strain(1:50) was significantly lower than that of serum induced by parental strain(＞ 1:800).Virulence test showed that M5-90Δbp26 strain was less virulent than parental strain.Conclusions M5-90Δbp26 has been successfully constructed.M5-90Δbp26 of Brucella melitensis has the characteristic of reduced virulence and has a potential as brucellosis candidate vaccine strain permitting serological discrimination between diseased and vaccinated livestock.

BACKGROUND:Due to the importance of pedicle adjacent structures, once the screw replacement appears a deviation, adjacent structures may be damaged, leading to extremely serious consequences. Although the security of screw placement in thoracic vertebral pedicle-rib complex is significantly greater than that of pedicle screws, the mechanics of the pedicle-rib complex at different cross-sectional areas of the screw are rarely reported.
OBJECTIVE:To observe mechanical property of different cross-sectional area screws in the middle and upper thoracic vertebral pedicle-rib complex.
METHODS:Five specimens of adult cadaveric thoracic spine (T 1-T 10 ) and adjacent rib segment (50-60 mm long) were used. The bone density of specimens was measured using difunctional bone density testing machine, and osteoporotic vertebral body was excluded. The position of the screws was detected with CT images. The maximal withdrawal force of the pedicle screw was measured with biomechanical force test machine.
RESULTS AND CONCLUSION:Thirty-eight specimens at normal bone density were implanted with 25 screws (5.5 mm), 25 screws (6.0 mm) and 26 screws (6.5 mm). Because the pedicle screws destroyed the pedicle-rib complex and perforated the vertebral body, we final y obtained the withdrawal force of 68 screws. The axial withdrawal force of pedicle screws at different diameters was (812.36±147.22) N, (868.64±160.48) N and (946.48±157.58) N, respectively. There were significant differences between the 5.5 mm screws and the 6.5 mm screws (P<0.05). Experimental findings indicate that, the pedicle screws (diameter>5.5 mm) are suitable in the middle and upper thoracic vertebral pedicle-rib complex due to strong internal fixation and clinical requirement.

Objective In order to improve the understanding of Kimura's disease,the clinical features and the pathological changes of 12 patients were analyzed.Methods Twelve cases with Kimura's disease ad- mitted to Qilu Hospital of Shandong University were retrospectively reviewed.Results All 12 patients were in relatively good condition and presented as subcutaneous nodules or swelling lymph nodes.Peripheral blood eosinophilia did not occur in 5 cases,4 out of 6 patients had high-level serum IgE.Biopsies were taken in all cases and the characteristic histological presentations were discovered.Only one patient developed pulmonary inflammation and acute myocardial infarction which were rare in Kimura's disease.Conclusions Definite di- agnosis of Kimura's disease mainly relies on biopsy.A patient with Kimura's disease can suffer from severe pulmonary and cardiac diseases,but the relationship between them should be studied further.

Objective To study the clinicopathologic features of brain tumors in patients with medically intractable epilepsy. Methods The clinical, radiologic and pathologic features of brain tumors in thirty-six patients with intractable epilepsy encountered during the period from 2008 to 2014 in the Epilepsy Center of Haidian Hospital were retrospectively reviewed. Results There were 18 males and 18 females in thirty-six patients. The mean age of seizure onset and disease duration were (14.05 ± 1.67) years and (10.04 ± 1.19) years respectively. The histological types of brain tumors included ganglioglioma (12/36, WHO gradeⅠ,1/36, WHO gradeⅡ), dysembryeplastic neuroepithelial tumor (2/36, WHO gradeⅠ), pleomorphic xanthoastrocytoma (1/36, WHO gradeⅡ), angiocentric glioma (1/36, WHO gradeⅠ), astrocytoma (4/36, WHO gradeⅡ), oligoastrocytoma (1/36, WHO gradeⅡ, 2/36, WHO gradeⅠ-Ⅱ), oligodendroglioma (1/36, WHO gradeⅠ-Ⅱ,1/36, WHO grade Ⅱ), cavernous hemangioma (4/36) and Sturge-Weber syndrome (1/36). Most of these tumors were located in temporal lobe (25/36, 69.4%). Patients were followed up for 0.5-7 years after operation. One patient was lost for follow up. Seizure outcome after the epilepsy operation revealed that 28 patients (77.8%) had Engel gradeⅠ, 4 patients (11.1%) had Engel gradeⅡ,2 patients (5.6%) had Engel gradeⅢ,1 patient (2.8%) had Engel gradeⅣ. Conclusion Brain tumors in patients with medically intractable epilepsy are almost low grade tumors of the nervous system. Focal cortical dysplasia is existed in most brain tissues from the epilepsy operation. Low grade tumors of the nervous system have close relation with focal cortical dysplasia in patients with medically intractable epilepsy. It is possible that the classifications of pathology diagnosis has connection with prognosis.