Pig copper zinc superoxide dismutase (CuZnSOD) is an important antioxidant enzyme. Some studies focused on the function of CuZnSOD gene, but the transcriptional regulation of the CuZnSOD gene is not yet fully elucidated. Therefore, the aims of the study were to determine the core promoter region and to explore its mechanism of transcriptional regulation. The 853 bp DNA sequence of 5'-flanking promoter was amplified by performing PCR. A series of CuZnSOD promoter fragments with gradually truncated 5'-end were produced by nested PCR and inserted into pGL3-Basic vector. The activities of the promoters were measured by the dual-luciferase assay system after transient transfection into the NIH/3T3 cells. The results demonstrated that there were 2 potential transcription start sites in the regions from initiation codon to -87 bp and -266 bp, respectively. The region from -383 bp to +67 bp in CuZnSOD gene promoter showed higher activity than other regions, and further deletion analysis demonstrated that the region from -75 bp to -32 bp contained an essential promoter sequence for pig CuZnSOD gene transcription. In addition, several potential transcription factor binding sites were predicted with bioinformatics method. These results suggest that these transcription factor binding sites may be involved in the transcriptional regulation of CuZnSOD gene.
Animals ; Cloning, Molecular ; Gene Expression Regulation ; Genetic Vectors ; Mice ; NIH 3T3 Cells ; Promoter Regions, Genetic ; Superoxide Dismutase ; genetics ; Swine ; Transfection

Objective The aim of this study was to investigate the relationship between phosphotyrosine interac-tion domain containing 1 ( PID1 ) gene and variation in intramuscular fat ( IMF ) content and the possibility to generate transgenic animals by testicular injection .Methods Expression vector pIRES2-acGFP-PID1 carrying pig PID1 gene was incubated with transfection reagents and injected into the testes of male New Zealand rabbits .We examined the F1 genera-tion by fluorescence detection , PCR, Western blotting and measuring the IMF content .The F1 generation gave birth to the F2 generation.Then we examined the F2 generation through detecting the positive rate and the IMF content .Results The exogenous PID1 gene and fluorescent protein gene were expressed at different levels both in the F 1 generation and the F2 generation, and the positive rates were 35.88%and 34.33%, respectively.The IMF content was significantly elevated (P<0.05) in the transgenic positive individuals compared with the negative ones and the control group , and the PID1 protein expression was similarly higher .Conclusions The results of this study demonstrate that PID 1 gene affects intramuscular fat content significantly .Moreover, the results of our analysis provide further evidence that transgenic animals can be gener -ated by testicular injection , and the exogenous gene can be inherited steadily .

Objective To investigate the expression profile of human genes in response to acute sodium arsenite treatment by cDNA microarray. Methods The RNA was purified from the L-02 cells without and with arsenite sodium induction for 2 hours, 15 hours and 24 hours, respectively. Results The hybridization patterns were different between every interval of arsenite induction. Expression of hCYR61 increased after 2 hours' induction, but decreased after 15 hours and 24 hours. Expression of metallothionein Ⅳ and Ⅲ elevated at the whole induction phase. HSP86 was up-regulated after 15 hours and 24 hours' induction, but it did not alter at two hours' induction. Conclusion When exposed to arsenite, the cells are under a meet-an-emergency situation to synthesize the most necessary protein and inhibit synthesis of unessential proteins.

Objective To investigate the expression and significance of activator protein-1 (AP-1)and matrix metalloproteinases (MMPs) in acute myocardial infarction (AMI) subjects. Methods Immunohistochemical techniques were used to detect the subunit of AP-1 (c-Jun), MMP-2 and MMP-9 in human AMI and normal heart tissue and the expressions of c-Jun and MMPs were measured with computer image analysis system. Results (1) There were expressions of c-Jun, MMP-2 and MMP-9 in normal heart tissue, mainly in myocardial cells and cardiac fibroblasts, and their expressions in AMI myocardial tissues were all significantly higher than those in normal myocardial tissues (P <0.05). (2) The level of MMP-9expression was significantly and positively correlated with c-Jun in AMI heart tissue (r=0.773, P < 0.01).Conclusions The expressions of AP-1 and MMPs increase in human myocardial infarction. These findings suggest that AP-1 transcription activation pathway and MMPs may play an important role in ventricular remodeling of myocardial infarction.

According to the designed specific primers of gene fragment based on the Salvia miltiorrhiza transcriptome data, with the method of reverse transcription polymerase chain reaction (RT-PCR), this study cloned full-length cDNA sequence of 1-hydroxy-2-methyl-2-(E)-butenyl-4-diphosphate synthase gene from Salvia miltiorrhiza bge.f.alba, this sequence is named as SmHDS and its GenBank registration number is KJ746807. SmHDS, 2 529 bp long, contains an ORF of 2 229 bp, encodes 742 amino acids, including 5' UTR 170 bp and 3' UTR 130 bp. Using bioinformatics software, having made a homology analysis of the obtained sequence, we can have a conclusion that SmHDS have a close genetic relationship with HDS of Salvia miltiorrhiza. Analysis result of prokaryotic expression revealed that in Escherichia coli, SmHDS expressed target proteins which in size are comparable with the protein predicted. Meanwhile, the 4 factors which can influence the protein expression were optimized, the 4 factors are inducing temperature, inducing time, IPTG concentrations and density of inducing host bacterium (A600). The optimal expression conditions of SmHDS were 30 degrees C until the A600 is 0.6, and add IPTG to a final concentration of 0.2 mmol x L(-1), and the induction time of 20 h. It provides theoretical basis for the further study of the function of 1-hydroxy-2-methyl-2-(E)-butenyl-4-diphosphate synthase in the biosynthesis of tanshinone compounds.

In this study, we demonstrated that immobilized fibronectin (FN) enhanced LAK activity, and that the enhanced LAK activity was completely abrogated by an anti-VLA-5 monoclonal antibody and RGD peptide. Fresh -spleen cells expressed VLA-4, VLA-6 and vitronectin receptor, whereas VLA-5 was expressed only on the spleen cells activated with IL-2. LAK cells showed increased adhesion to immobilized FN compared with that to control BSA, and the increased adhesion of LAK cells to immobilized FN was inhibited by anti-VLA-5 monoclonal antibody. Conjugate-formation assay showed that the LAK cells cultured on immobilized FN bound to target cells more efficiently than the control LAK cells, and that anti-LFA-1 monoclonal antibody inhibited the LAK-target cell binding. Immobilized type IV collagen and laminin, as well as FN, enhanced LAK activity. All these results suggest that the interaction of inte-grins expressed on LAK cells with extracellular matrix proteins act as co-stimulator for the enhancement of LAK activity , and that anchorage is necessary for full activation of LAK cells.

AIM:To investigate the effects of ox-LDL on TLR2 and TLR4 expression and production of TNF-?,IL-10,IL-12,NO and MDA in macrophages and to observe intervention effect of GW1929 in above procedure.METHODS:The mouse peritoneal macrophages were pretreated with ox-LDL(50 mg/L,100 mg/L)and GW1929(20 ?mol/L)respectively for 24 h.The concentrations of MDA,NO-2/NO-3,TNF-?,IL-10 and IL-12 in the culture fluid were detected.Flow cytometry was used to observe TLR2 and TLR4 expressions after the mouse peritoneal macrophages were pretreated with ox-LDL(50 mg/L)and GW1929(20 ?mol/L)respectively for 6 h,12 h,and 24 h.RESULTS:The concentrations of MDA,NO-2/NO-3,TNF-? and IL-10 in ox-LDL(50 mg/L,100 mg/L)group were higher than those in control and GW1929 group obviously,but the concentrations of above index in ox-LDL(50 mg/L,100 mg/L)+GW1929 group were lower than those in ox-LDL(50 mg/L,100 mg/L)group apparently.No IL-12 in every group was detected.Expressions of TLR-2 in ox-LDL+GW1929(6 h,12 h,24 h)group were lower than those in ox-LDL(6 h,12 h,24 h)group respectively.TLR-4 expressions in ox-LDL+GW1929(12 h)were lower than those in ox-LDL(12 h)apparently.CONCLUSION:ox-LDL up-regulates TLR2 and TLR4 expressions and promotes the production of ROX,NO,TNF-? and IL-10 in macrophages.GW1929 is capable of inhibiting the above ox-LDL effects.

Objective: To understand the parenting styles of junior high school students in Beijing, to explore the relationship between parenting styles with students academic performance and the role of mental health in this association. Methods: From October 2019 to January 2020, a stratified random cluster sampling method was used to select 5 196 students in the first and second grades of 11 public junior high schools in urban and rural areas in Beijing, and Slort Egna Minnenar Barndoms Uppfostran-China,Middle School Students Mental Health Scale-60 and self designed questions were used in the questionnaire survey. Results: The average score of parenting style rejection dimension was (9.44±3.88), and the average score of emotional warmth dimension was (19.85±5.66). The average score of comprehensive score of students mental health was ( 1.92 ±0.73). Mental health score of girls was higher than boys (1.97±0.74)(1.87±0.71), and the differences were of statistical significance( t=5.06, P <0.01). Parental rejection and emotionally warm parenting styles were significantly correlated with students academic performance （ β =-0.54，1.15， P <0.01). Mental health played a negative moderating role between parenting style and students academic performance. The better students mental health were, the greater the influence of parenting style on academic performance. Conclusion Parenting style is related to children s academic performance. Parents should give their children more affirmation, encouragement and praise, in order to improve their academic performance.

Key healthcare resources are doubtlessly critical in emergency management. How to balance the supply and demand, use limited resources scientifically and rationally, and maintain a healthy public service system, have emerged an important challenge in emergency command. Based on the theory of supply-demand balance of public goods, the article took Wuhan as an example to analyze and establish a model on key healthcare resources allocation, in order to provide reference and evidence for global health governance and other similar public health emergencies.

Upholding the overall leadership of the Party is an important cornerstone for promoting development of public hospitals. Party building leadership for high-quality development of public hospitals must adhere to the value orientation of public welfare, establish a modern hospital management system, and promote the mutual promotion and integration of both Party building and medical service. The practice path of Party building in leading the high-quality development of public hospitals in a tertiary hospital won the support of patients, employees and medical students at large, gave full play to the leading role of the Party, and made breakthroughs in terms of medical service, discipline construction, talent training and internal management quality improvements. By strengthening top-level design and institutional system construction, the hospital has effectively transformed the political and organizational advantages led by Party building into an endogenous momentum for the reform and development of public hospitals. The practice solved such important questions as to the beneficiaries of the high-quality development of public hospitals, whom to depend on and how to govern, hence providing reference for further promoting the high-quality development of public hospitals in China and realizing the deep integration of Party building and medical service.