AIM: To get large amounts of pure antigens to raise specific antibodies and to perform quantifications.METHODS: CYP2B6 (cytochrome P) cDNA fragments was ligated into BamHI restricted PGEX-3b to generate recombinants PGEX/2B6. We identified recombinants PGEX/2B6 by EcoRI digestion. The expression of fusion proteins were induced by adding isopropyl-thiogalactoside(IPTG). Several clones showed high-level expression of fusion proteins. Insoluble proteins was isolated from the bacteria and the fusion proteins was recovered and purified from a preparative (2mm) SDS-PAGE. The polyarcrylamide gel containing the fusion proteins glutathione S-transferase(GST-2B6) were used to immunize BALB/C mice from which polyclonal ascites fluid was prepared. The purified fusion proteins GST-1A1(GST fusion protein of CYP1A1 cDNA246～386aa expressed in this library ,purified by preparative SDS-PAGE), GST-2B6 were used to test the specificity of 2B6pAb. RESULTS:Fusion proteins constructed between GST and CYP2B6 was expressed in Escherichia coli DH5?. Mouse antibodies are raised against the fusion proteins GST-2B6. 2B6pAb was fond to be specific antibody.CONCLUSION:Recombinant PGEX/2B6 were constructed and purified fusion proteins GST-2B6, and specific 2B6pAb were obtained.

To increase the conclusion rate of scientific research projects and prevent the phenomena of fine start and poor finish,high input and low output,and heavy application and light management therein,by calculating the conclusion rate of the research projects of all levels expired longitudinally undertaken by Fujian Medical University from 2010 to 2012,this thesis analyzes the root cause for the delay of research projects starting from the three parties which sign the research project contract,namely,the competent authority of science and technology,the regulation department of scientific research,and the research group; meanwhile,it puts forward the countermeasures including perfecting the management system of scientific research,implementing effective secondary Strengthen the team management,management of colleges and universities,and strengthening the building of scientific research management team,etc.,in the hope of providing reference for the increase of the conclusion rate of research projects in institutions of higher learning.

Objective:To evaluate the effects of crown lengthening surgery by apically repositioned flap for the treatment of the teeth with inadequate width of attached gingiva. Methods:17 teeth with inadequate width of attached gingiva were included. The gingiva flap was repositioned apically. The distance between gingival margin to the root surface was recorded, the width of attached gingiva and other indicators were compared before and 6 months after surgery. Results:Edge of the defects the 17 teeth was below gingiva margin before surgery, root surface was exposed to the coronal gingival margin 6 months after surgery in 16 of the 17 cases (94. 11%). Api-cally displacement distance of the gingival margin was (3. 88 ± 0. 49) mm. The width of attached gingiva before and 6 months after sur-gery was (2.35 ±0.61) mm and (2.65 ±0.49) mm respectively(P>0.05). Conclusion: Crown lengthening with apically reposi-tioned flap surgery can effectively expose the root surface and create conditions for restoration of dental defects for teeth with inadequate attached gingiva, and can prevent over removal of the keratinized gingiva.

BACKGROUND: With development of modern medical sciences, the out comes after meningioma surgery cannot be fully assessed using morbidity,livability and mortality merely. Quality of life (QOL) is a health-related multivariable index, which provides integrated inf ormation associated with physiological,sychological and social adaptation status of the patients to doctors. Assessment of the QOL in meningioma patients may be one of a relatively good method for guiding surgery and reducing reoccurrence of tuOBJECTIVE: To evaluate the QOL in meningioma patients and then to provide relevant data for assessing the outcomes of meningioma surgery comprehensively.DESIGN: It was a single-sample investigation. SETTING: Neurosurgery Department, Second People's Hospital of Wuxi City of Nanjing Medical University.PARTICIPANTS: Totally 147 patients underwent meningioma surgery in the Neurosurgery Department of the Second Affiliated Hospital of Wuxi City affiliated to Nanjing Medical University between January 1995 to January 2001 were selected, 61 males and 86 females, with a male/female ratio of 2:3 and an age ranging from 5-77 years old, in average of 43 years old, and the median was 43 years old.METHODS: Based on World Health Organization Quality of Life 100item questionnaire (WHOQOL-100) and Karnofsky Performance Scale (KPS), we designed a questionnaire on QOL in patients with meningioma.Outcomes of the transverse investigation on 147 meningioma patients were compared with healthy controls. Surgery-related QOL curve was used for identifying the threshold satisfactory point of QOL. In addition, with selfcontrol, influence of surgery on QOL was assessed.MAIN OUTCOME MEASURES: Relationship between QOL and clinical histories, radiological analysis, operation grading, histological properties and reoccurrence were investigated.RESULTS: All of the 147 patients entered the statistical analysis. ①In patient with meningioma, Cronbach's coefficient was 0.9521 and the correlated coefficient was 0.8685, which suggested that QOL questionnaire for patients with meningioma had a good reliability and validity. ②QOL ≥70 represented that the patient was satisfied with the QOL. ③In patients underwent surgical treatment, their physiological functions, self-help and self-care except for psychological functions were significantly improved (P ＜ 0.001 ).CONCLUSION: Surgical treatment enables patients to get a satisfied QOL. QOL questionnaire for patients with meningioma is helpful to provide relevant data for meningioma operation.

Three-orientation medical talent training mainly include humanistic quality cultivation, professional quality and physical and mental quality. With professional ability as the guidance of profes-sional quality education requires students not only to grasp skilled medical technology, but also have the ability to enhance the medical technology. Based on the orientation of training target, this research mainly from habit set, authority set, set the multitude and empirical mind-set four aspects in this paper, the critical thinking, and put forward problems from finding problems, and expounds the innovative thinking of concise problem.

Objective To explore the reversion effect of Gemcitabine-resistance A549 cell (A549/Gem) by silencing CXCR4.Methods A549 cell was induced by continuous stepwise exposure to Gemcitabine in order to obtain Gemcitabine-resistance A549 cell ( A549/Gem) in vitro.The CXCR4 expressions level of A549 and A549/Gem were detected by Quantitative RT-PCR ( RT-qPCR) and Western blot analyses.The CXCR4 shRNA vector was transfected into the A549/Gem cell by targeting silence CXCR4.Furthermore, MTT assay was used to explore the IC50 and RI in A549, A549/Gem and A549/Gem-CXCR4 cells.Moreover, Western blot analysis was performed to detect the expressions of phospho-JNK, phospho-p38 and phospho-ERK 1/2 in A549, A549/Gem and A549/Gem-CXCR4 cells.Results Gemcitabine-resistance A549 cell ( A549/Gem) was successful constructed by using continuous stepwise exposure to Gemcitabine in vitro.The expression level of CXCR4 was up-regulated in A549/Gem cell than in A549 cell.The CXCR4 shRNA vector could significantly decrease CXCR4 expression in A549/Gem cell.The IC50 values of Gemcitabine in A549, A549/Gem and A549/Gem-CXCR4 cell were (0.08 ±0.01)μmol/L, (14.01 ±0.21)μmol/L and (1.84 ±0.61)μmol/L, respectively.The RI value of Gemcitabine was (127.12 ±12.28) in A549/Gem cells, while the value of RI was (27.3 ±0.98) in A549/Gem-CXCR4 cells.The expression level of phospho-JNK, phospho-p38 and phospho-ERK 1/2 were also markedly inhibited in A549/Gem-CXCR4 cell than in A549/Gem cell.Conclusion CXCR4 is up-regulated in A549/Gem cell.Targeting silence CXCR4 can successfully reverse drug-resistance of Gemcitabine in A549/Gem cells, which hints CXCR4 is associated with lung cancer radiation therapy as an effective molecular target.

Objective To use evidence-based nursing on patients with artificial airway to the practice of stomach tube, and to evaluate the self made guidance for difficult gastric tube placement in patients with artificial airway.Methods Forty patients with artificial airway and were difficult to put the tube, and admitted to Department of Critical Care Medicine of Harrison International Peace Hospital Affiliated to Hebei Medical University from April to December in 2016 were enrolled as observation group. Through the evidence-based nursing strategy, the related literatures at home and abroad were collected to search clinical evidence and formulate and implement the nursing program, the gastric tube was inserted into the stomach tube under the direct vision of the laryngoscope. Thirty-six patients of difficult gastric tube placement with artificial airway straightly under the direct vision of the laryngoscope from August 2015 to March 2016 were retrospectively analyzed as the control group. The success rate of first catheterization, indwelling time, throat edema and bleeding of pharyngeal mucosa were compared between the two groups.Results All patients were enrolled in the final analysis. In the control group, 28 patients were successfully placed once, while 8 failed. Only 1 patient in observation group failed to catheterize, and successful placed after symptomatic treatment, the one-time success rate of catheter was significantly higher than that of control group (97.5% vs. 77.8%), and catheter time was significantly shortened (minutes: 4.8±1.2 vs. 5.1±1.0), the difference was statistically significant (bothP < 0.05). There were 2 patients with laryngeal edema in the control group and 4 patients of pharyngeal mucosal hemorrhage. In the observation group, there was no laryngealedema and laryngeal edema occurred in the patients with laryngoscopy, and only 1 patient had a hemorrhage of pharyngeal mucosa.Conclusion Using self made guiding device can effectively insert the difficult gastric tube in patients with artificial airway, and increase the one-time success rate of intubation, shorten the catheter time, and have a trend in reduce complication as compared with traditional gastric tube placement.

Objective To explore the quantity and ultrastructure of dopaminum (DA) neuron in nigra-striatum, pathological basis of the successful animal model of cerebral palsy (CP) by hyperbilirubinemia, to provide the evidence for clinical therapy and rehabilitation. Methods Thirty homozygous newborn rabbits (2~5 d) were randomly divided into 2 groups: a control group(n=10) and a model group(n=20). The rabbits in the control group were administrated bilirubin intraperitoneally, with a total dose of 300 mg/kg, while those in the model group were administrated with saline at the same dose. All the animals were fed well for 45 days until they were sacrificed for observation of the ultrastructure of brain tissue and quantification of DA neurons. Results The ultrastructure of nigra-striatum of the control group demonstrated significant changes; The quantity of nigra-striatum DA neurons of the control group decreased. Conclusion The ultrastructure of nigra-striatum of the control group has had obviously changes. The nigra-striatum DA neuron of CP rabbits was impaired and the quantity of nigra-striatum DA neuron of CP rabbits was reduced.

ObjectiveTo study change of aminoacid content in the brain of newborn rabbits with bilirubin encephalopathy. MethodsNewborn rabbits (3-5 days after birth) with bilirubin encephalopathy were induced by administration intra peritoneally with bilirubin (200mg/kg). Aminoacid content in brain were analysed by Hitachi 835 50 type aminoacid analysis instrument.ResultsThe content of glutamate and aspartate in brain tissue with bilirubin encephopathy decrease; GABA, alanine, tyrosine, valine and pheny lalanine increase. ConclusionsThe change of content of neurotransmitters in brain indicates that excitotoxicity of EAAs may contribute to the bilirubin encephalopathy. The increase of non neurotransmitters level proves that bilirubin may inhibit the biologic composition of protein in brain.

ObjectiveTo explore the noxious effect and the extent of damage in nigra-striatum neuron of bilirubin.Methods30 newborn rabbits were divided into 3 groups randomly:control group(group C,12 rabbits) which were injected normal salt intraperitoneally,model group 1(group N1,12 rabbits) which were injected bilirubin 100mg/kg intraperitoneally, model group 2(group N2,12 rabbits) which were injected bilirubin 200mg/kg intraperitoneally. All the rabbits in group C and 6 rabbits in group N1(group N1a) and group 2(group N2a) were killed 6 hours after injection.Other 6 rabbits in group N1(group N1b) and group 2(group N2b) were killed 16 hours after injection. The quantity of neuron in nigra-striatum were determined,and the changes of ultrastructure were observed by electron microscope.ResultsThe neuron in nigra-striatum in group N2b were decreased compared with group C and group N1a(P<0.05),as well as with group N1b and N2a. The ultrastructure of the neuron was changed.ConclusionsThe changes in utrastructure and the quantity of nigra-striatum neuron were associated with the concentration and time of exposure under bilirubin. It is consisted with the changes of ultrastructrue and quantity in nigra-striatum.