Objective:To amplify full-length expressing sequence of human granzyme B(GrB) and perforin(PFP) firstly,then to construct a co-expression vector of pVAX1-PIG and to analyze the expression of human GrB and PFP in Hep-2 cells.Methods:The human PFP and GrB expressing sequences were amplified by RT-PCR.The expression vector pVAX1-PIG was constructed.The recombinant vector was transfected into human Hep-2 cells and their expressions were detected by RT-PCR and indirect immunofluorescent antibody assay.Results:The GrB and PFP cDNA fragment were cloned into the vector of pVAX1 in the right direction.The target proteins were detected in the transfected Hep-2 cells.Conclusion:The vector of pVAX1-PIG was constructed successfully and expressed in Hep-2 cells line.The perforin/granzyme B protein can induce cell apoptosis.These results provide some foundation in gene therapy for tumor by making use of PFP and GrB gene.

Antigens, Bacterial ; genetics ; metabolism ; Bacterial Proteins ; genetics ; metabolism ; Child ; Child, Preschool ; DNA, Bacterial ; genetics ; Gastritis ; pathology ; Gene Expression Regulation, Bacterial ; Helicobacter Infections ; microbiology ; pathology ; Helicobacter pylori ; genetics ; Humans ; Immunohistochemistry ; Peptic Ulcer ; microbiology ; pathology ; Polymerase Chain Reaction

Fifty five patients with alcohol induced-mental disorder (study group) and 43 local inhabitants without history of alcohol abuse (control group) were surveyed with family environment scale (FES-CV) and generic quality of life inventory-74 (GQOLI-74). The total score and the scores of all dimensions except material life in GQOLI-74 of study group were significantly lower than those of control group(P ＜0. 05). Compared with control group, the scores in FES of study group were lower for factors of cohesion, expressiveness, active-recreational orientation, moral-religious emphasis and organization in the patient's family, while the scores for conflict and control were higher( P ＜ 0. 05 or P ＜ 0. 01 ). The results indicate that family environment is closely correlated with quality of life in patients with alcohol-induced mental disorder, and family therapy would improve their quality of life.

Nitric oxide(NO)is a vital signal messenger in human and generated by nitric oxide synthase including endothelial nitric oxide synthase(eNOS), inducible nitric oxide synthase(iNOS)and neuropathic nitric oxide synthase(nNOS). NO has important regulatory function on the immune system, nervous system and many other physiological systems. Endogenous NO can enhance the function of the vascular system and endothelial cell survival, and inhibit platelet accumulation and leukocyte infiltration. Excessive production of NO may damage tissues by cytotoxic and cytostatic effects. Thus, NO plays a dual role in physiology and pathology. This paper gives a brief review on regulatory effects of NOS-NO system by traditional Chinese medicine during the recent five years, so as to provide some clues or references for the treatment of related diseases and scientific evidene for reasonable and effective clinical application.

BACKGROUND:In clinical application,the therapeutic effect of transcranial magnetic stimulation depends on the ability to accurately target the areas of the brain that need to be stimulated.In recent years,with the development of neuronavigation systems,mobile augmented reality technology,and the new methods of processing magnetic resonance imaging(MRI)data,the accuracy of stimulus target localization and the optimization of target selection are expected to improve further. OBJECTIVE:To review the principle of MRI-based image navigation and its application in transcranial magnetic stimulation and summarize the roles of different modal MRI data analyses in guiding the selection of target areas for transcranial magnetic stimulation. METHODS:An online computer search for relevant literature was performed in PubMed,CNKI database and WanFang database,with the keywords"transcranial magnetic stimulation,coil positioning,neuronavigation,augmented reality,magnetic resonance,theory."Finally,63 documents were included for review. RESULTS AND CONCLUSION:Among the traditional methods of positioning transcranial magnetic stimulation coils,the"5 cm rule"and the international electroencephalogram 10-20 positioning method are the most commonly used.These methods have the advantages of simplicity and economy,but they rely too much on the operator's experience and there were technical differences between operators.The neuronavigation system,which is based on stereotactic technology,is the guiding method for positioning transcranial magnetic stimulation coils with the highest visual degree and accuracy.It achieves visual positioning through MRI data acquisition,3D brain reconstruction,head model registration and stereogeometric positioning.It has high application value in clinical treatment and scientific research,but it cannot be promoted in medical institutions due to its high cost.For various medical institutions,mobile augmented reality is a cost-effective and efficient alternative to the neuronavigation system,which achieves visual positioning of brain tissue under the scalp through MRI data acquisition,2D/3D image construction,virtual image and real brain image superposition.It has the advantages of directly visualization and low cost,and is expected to be popularized and applied in primary medical units.Although the superiority of clinical efficacy of visual coil positioning over the electroencephalogram 10-20 localization strategy has not yet been fully demonstrated,with the progress of brain MRI data analysis,visual positioning is expected to further optimize the target selection strategy of transcranial magnetic stimulation therapy and to improve the response rate and individuation degree of transcranial magnetic stimulation treatment.This is a promising and challenging research direction in the future.

ObjectiveTo explore the potential mechanism of Dingchuan Granule （定喘颗粒， DG） in the treatment of respiratory syncytial virus （RSV） pneumonia. MethodsA total of 60 male Sprague Dawley （SD） rats were randomly divided into control group， model group， ribavirin group， DG low-dose group， DG middle-dose group， and DG high-dose group， with 10 rats in each group. Except for the control group， rats were administrated with RSV via intranasal drip. After model establishment， the DG low-， middle-， and high-dose groups were administrated via oral gavage with DG at 3.47， 6.93， and 13.86 g/（kg·d） respectively， while the ribavirin group was administrated via oral gavage with ribavirin at 15.75 mg/（kg·d）. The drug was given once daily for one week. The rats in the control group and the model group were not given any drug， only subjected to the grasping action. Twenty-four hours after the last administration， the pathological changes of lung tissues were observed and scored using HE staining. The levels of serum inflammatory factors， including tumor necrosis factor-α （TNF-α）， interleukin-1β （IL-1β）， and interleukin-6 （IL-6）， were detected by colorimetry. The protein levels of nuclear factor （erythroid derived 2）-like 2 （Nrf2）， Kelch-like ECH-associated protein 1 （Keap1）， heme oxygenase 1 （HO-1）， and NAD（P）H quinone dehydrogenase 1 （NQO1） in lung tissues were measured by Western Blot. The RSV load as well as the gene expression levels of Nrf2， Keap1， HO-1， and NQO1 in lung tissues were determined by qRT-PCR. The level of reactive oxygen species （ROS） in rat lung tissues was detected using chemiluminescence. The levels of glutathione （GSH） and malondialdehyde （MDA） in rat lung tissues were measured by a microassay. ResultsCompared with the control group， other groups had significant increases in pathological score of lung tissue， RSV load， levels of ROS， MDA， serum TNF-α， IL-1β， and IL-6； decrease in GSH level， increases in expression level of Keap1 protein and its mRNA in lung tissue， and significant decrease in levels of Nrf2， HO-1， expression level of NQO1 protein and its mRNA （P＜0.05）. Compared with the model group， all the above-mentioned indicators in the DG low-， middle-， and high-dose groups and the ribavirin group were improved to varying degree （P＜0.05）. The levels of serum TNF-α， IL-1β， and IL-6 in rats of DG dose groups showed a dose-dependent pattern， the DG high-dose group exhibiting the best effect （P＜0.05）. The DG high-dose group was superior to the DG low- and middle-dose groups in reducing the levels of ROS and MDA， and increasing the level of GSH in lung tissues （P＜0.05）. The DG high-dose group and the ribavirin group had better effect than the DG middle-dose group in reducing the RSV load （P＜0.05）. The DG high-dose group was superior to the ribavirin group in improving the protein levels of Nrf2， Keap1， HO-1， and NQO1 （P＜0.05）. ConclusionDG could inhibit oxidative stress by regulating the Nrf2/Keap1/HO-1/NQO1 signaling pathway to improve pulmonary inflammation and treat RSV pneumonia， with the DG high-dose group showing the best effect.