Objective To determine the effect of Vibrio vulnificus cytolysin (VVC) inducing ap-optosis in human umbilical vein endothelial cell(HUVEC) and its possible mechanism. Methods The en-tire vvhA gene that encoding VVC from V. vulnificus strain GTC333 was amplified by PCR and sequenced af-ter T-A cloning. E. coli BL21DE3pET-42a-vvhA, a prokaryotic expression system of the vvhA gene, was then con-structed. Ni-NTA affinity chromatography was applied to purify the target recombinant protein rVVC, and SDS-PAGE plus Bio-Rad Agarose Image Analyzor were used to measure the output of rVVC and to determine the purity of rVVC extract. The activity of rVVC dissolving rabbit erythrocytes was detected by hemolysis test. DPNH chromotometry and TphBNa chromotometry were performed to examine the contents of LDH and K+ in the supernatants of rVVC-treated HUVEC cultures, respectively. The effect of rVVC inducing apepto-sis of HUVEC was detected by flow cytometry, rVVC was labeled with FITC and the location of FITC-labe-ling rVVC in HUVEC was observed by laser canfocal microscopy. Results The cloned whA gene had 96.09% and 98.26% similarities of nucleotide and amino acid sequences compared to the corresponding se-quences in GenBank. rVVC, with a dosage of 1 μg/ml, could dissolve rabbit erythrocytes (P<0.01). 10 μg/ml rVVC was able to promote the increases of K+ content (P<0.01) but no change of LDH content could be found in the cell supernatants. HUVEC was apoptotic after the cell was treated with 1～100 μg/ml of rVVC for 2 h. In the 5～240 min duration of co-incubation of FITC-labeling rVVC and HUVEC, the rV-VC gradually moved from surface to inner side of the membrane and then entered the cytoplasms. When FITC-labeling rVVC treated HUVEC for 30 min, most of the rVVC was found to be intracellular location. Conclusion rVVC has cytolytic activity. VVC has an ability to enter HUVEC and causes injury of HUVEC via inducing apoptosis, which may be the major pathogenic mechanism of VVC.

As an advanced conformal therapy, Intensity-modulated radiotherapy (IMRT) can increase the gain ratio of radiotherapy and improve the local control of the tumor. This paper applies genetic algorithm to optimization of IMRT inverse planning. The algorithm model and simulation result are introduced.

ObjectiveTo assess the current situation of postgraduates knowledge about medical research design and optimize the curriculum setting of research design.MethodsAn investigation was carried out in the postgraduates of 2011 using questionnaires in a medical university..The questionnaire ineluded basic information of participants and cognition of basic methods of research design.ResultsA total of 473 postgraduates participated in the investigation.Among them,311 systematically learned medical statistics before enrollment,and 275 ( 58.14％ ) once participated in scientific researches.Most of them ( ＞80％ ) knew about the 10 basic methods of research design listed in the questionnaire,but only a few of them were familiar with them,and some even didn't know about the methods.ConclusionWe should pay attention to the culture of scientific research thought in statistical design,strengthen the practice of research design teaching,and the curriculum of research design should be led into undergraduate course.

By using the fuzzy min-max neural network, the quantitative structure-activity relationship (QSAR) of mutagenicity is studied. With the established QSAR model, the mutagenicity is predicted and the results showed that QASR is superior to linear-regression model. Further discussion on the models and the results is presented in this paper.
Algorithms ; Cluster Analysis ; Fuzzy Logic ; Models, Chemical ; Neural Networks (Computer) ; Quantitative Structure-Activity Relationship

Conformational restriction has become one of the important strategies for modification and optimization of lead compounds in structure-based drug design. In this review, we select some recent practical examples from literature to present applications of conformational restriction in structure-based drug design. By introducing fused rings, macrocycles, spiral rings, cyclopropyl groups, bridging rings, methyl groups into molecules, we can find that the conformational restriction strategy is widely used in improving pharmacokinetic properties, enhancing the activity and selectivity, and increasing the novelty of compounds. By using these conformational restriction strategies, the drug-like properties of lead compounds can be significantly improved. This paper can help to provide theoretical guidance and practical experience for innovation in drug design and development.

Objective To observe the change in IL-1β,IL-13mRNA expression in drowning rat lungs and serum,so as to investigate the significance of IL-1β and IL-13 mechanism in the development of drowning.Methods SD rats were randomly divided into control group,drowning group.Then using TaqMan probe method to determine the expression of IL-1β and IL-13 mRNA in Right lower lobe of lung tissue and the serum of right ventricle,which were extracted respectively from each group of rats.Results (1) The lung tissue morphological changes:Typical appearance signs and anatomy of drowning group meet ante-mortem drowning feature.(2) The expression of IL-1β,IL-13 in lung tissue:compared with the control group,the expression of IL-1β and IL-13 were slightly decreased,which has no statistical significance.(3) The expression of IL-1β and IL-13 in serum:compared with the control group,the expression of IL-1β and IL-13 were significant increased,both of which has statistical significance.Conclusion (1)The expression of IL-1β and IL-13 were decreased in lung tissue may be due to drowned rats present compensatory anti-inflammatory response syndrome which causes immune incompetent performance.(2) The expression of IL-1β and IL-13 were significant increased in serum may be relate to drown stress and drowning associated acute lung injury after traumatic stress.

ObjectiveTo investigate clinical features and outcomes in acute ischemic stroke patients with remote symptomatic intracranial hemorrhage (sICHr) after intravenous thrombolysis.MethodsThe acute ischemic stroke patients with sICHr after intravenous thrombolysis therapy were enrolled retrospectively.The clinical data were collected and the related literature was analyzed and summarized.ResultsA total of 6 acute ischemic stroke patients with sICHr were enrolled, including 4 males.Three patients had a history of using antiplatelet agents, 2 with atrial fibrillation, 4 with hypertension, 3 with previous stroke history, 4 with smoking history, and 4 had sICHr at 2 h after intravenous thrombolysis.Of the 14 hemorrhagic foci (except in the infarct areas), 10 were in the cerebral cortex.Three patients died within 1 week, and 1 was in a persistent vegetative state.Conclusions SICHr after intravenous thrombolysis in patients with acute ischemic stroke is mainly located in the cerebral cortex.The outcomes in acute ischemic stroke patients with SICHr after intravenous thrombolysis are poor, and the mortality is high.

OBJECTIVETo establish individualized prenatal diagnosis program for families affected with Duchenne/Becker muscular dystrophy (DMD/BMD) and different clinical background using a variety of methods.

METHODSMultiplex ligation-dependent probe amplification (MLPA) was performed on 50 patients suspected for DMD/BMD. For single exon deletions of the DMD gene, PCR was used for validating the results. For those without any deletion or duplication, Sanger sequencing was used to screen for DMD gene mutations in the children and their mothers. Prenatal genetic testing was provided to female carriers using chorionic villus, amniocentesis or cord blood samples. To ensure the accuracy of diagnosis, all prenatal specimens were also subjected to linkage analysis.

RESULTSAmong the 50 patients with DMD/BMD, 23 harbored large deletions, 11 only had single exon deletions, 10 harbored duplications, and 5 had small scare mutations. No mutation was detected in one family. For 37 women undergoing prenatal diagnosis, 10 fetuses were identified as affected males, 6 were female carriers, while 21 were not found to carry any mutation. Testing of creatine kinase was consistent with the results of prenatal diagnosis. For a patient harboring exon 51 deletion, the same mutation was found in a fetus but not in their mother. The proband and fetus had inherited the same haplotype, which suggested that the mother probably has germline mosaicism for the mutation.

CONCLUSIONApplication of individualized methods for analyzing pregnant women with different clinical background can minimize the risk for giving birth to further children affected with DMD/BMD.

Adolescent ; Child ; Child, Preschool ; DNA Mutational Analysis ; Exons ; Female ; Humans ; Male ; Multiplex Polymerase Chain Reaction ; Muscular Dystrophy, Duchenne ; diagnosis ; genetics ; Mutation ; Pedigree ; Pregnancy ; Prenatal Diagnosis

Ethical sensitivity is a behavioral concept emerging in recent years, which has attracted high attention from all sectors of society. Ethical sensitivity not only affects the clinical decision-making, doctor-patient relationship and medical quality of medical staff, but also provides the possibility to identify, predict and solve ethical problems in clinical. Combined with previous studies, this paper analyzed the current situation of ethical sensitivity of medical personnel in China, and took pediatric medical staff as an example to analyze the factors affecting the ethical sensitivity of medical staff from clinical practice and related theories, including age, working years, occupational categories, education level, work environment, empathy ability, etc. At the same time, this paper analyzed the causes of the ethical sensitivity of medical staff from the perspective of clinical practice and related systems, and put forward corresponding countermeasures, so as to improve the ethical sensitivity of medical staff and help to build a harmonious doctor-patient relationship.

Ethical sensitivity is a behavioral concept emerging in recent years, which has attracted high attention from all sectors of society. Ethical sensitivity not only affects the clinical decision-making, doctor-patient relationship and medical quality of medical staff, but also provides the possibility to identify, predict and solve ethical problems in clinical. Combined with previous studies, this paper analyzed the current situation of ethical sensitivity of medical personnel in China, and took pediatric medical staff as an example to analyze the factors affecting the ethical sensitivity of medical staff from clinical practice and related theories, including age, working years, occupational categories, education level, work environment, empathy ability, etc. At the same time, this paper analyzed the causes of the ethical sensitivity of medical staff from the perspective of clinical practice and related systems, and put forward corresponding countermeasures, so as to improve the ethical sensitivity of medical staff and help to build a harmonious doctor-patient relationship.