Objective:To construct the extracellular region of the human TRAIL cDNA expression vector and express and purify the extracellular region of the TRAIL protein. Methods: The mRNA of TRAIL was extracted from CD3 activated normal human PBMC and used as a template for reverse transcription. After PCR amplification, a 730 bp fragment including extracellular region was obtained and cloned into pGEX-2T.The recombinant vector was named pGEX/TRAILex. The pGEX/TRAILex vector was transformed into E.coli DH5a. After IPTIG induced at lower temperature, the collection of the sonicated extract was purified by using the GST agarose 4B. The purified fusion protein was identified by Western blotting with anti-TRAIL McAb.Results:The pGEX/TRAILex was constructed. After IPTG induced,a high level expression of the extracellular region of the TRAIL protein was obtained, SDS-PAGE analysis showed that the recombinant E. coli could express a 54 kD GST fusion protein which accounted for about 28% of the total cellular protein. The study of solubility of expression protein indicated that GST-Tex was expressed predominantly in the soluble form.The purified production was obtained 2.2 mg/L of culture media and the purity of the GST-Tex was more than 95%. GST/TRATLex protein could be recognized by anti-TRAL McAb in Western blot. Conclusion:The expression of recombinant extracellular domain of the human TRAIL protein may be useful for the study of biological functions of TRAIL and it's biotheraphy in tumor.

Objective To identify genomic DNA of Schistosoma ?. Methods Amplification of genomic DNA by the random-amplified polymorphic DNA polymerase chain reaction (RAPD-PCR) with 10-base pair was used. The 50 worms were collected from rabbits infected with cercariae of S.? and Schistosoma japonicum(S.j.) respectively. RAPD-PCR were performed on PCR-2400 according to the manufature's instruction. And 29 primers were adopted from Operon Company. The samples were run on 1.4% sepharose. Results RAPD fragments produced were various in quantity(4-12 bands)and size(0.5-5.2Kb) in S.? and S.j. , most of about 224 bands produced with 27 different primers were common, but 6 differential bands produced with 2 primers (J 01 CCCGGCATAA and L 12 GGGCGGTACT) of the 29 primers were found, 4 and 2 of the 6 differential bands seen in the S.? and S.j. respectively. Conclusion These specific fragments found in S.? and S.j. may be used as molecular markers for the identification of S.? and S.j.

Objective To explore the gray matter changes in aggressive patients with schizophrenia,and the relationship between the gray matter and aggression in patients.Methods Eighteen aggressive patients with schizophrenia (SZ1),18 age-and gender-matched un-aggressive patients with schizophrenia (SZ2) and 18 normal controls (NC) were enrolled in the study.Then a 3.0 T magnetic resonance imaging (MRI) scan was conducted for each participant.The voxel-based morphometry (VBM) approach and the Chinese version of Buss & Perry aggression questionnaire (B&P) were used to explore imaging data and to assess the aggression,respectively.Results Compared with NC,patients with schizophrenia showed changes in gray matter volume (GMV) in the frontal,temporal and the occipital lobes (P＜0.05,AlphaSim corrected).Compared with SZ2,SZ1 showed increased GMV in the right supramarginal gyrus,right postcentral gyms,bilateral insula and orbito-frontal gyri (P＜0.05,AlphaSim corrected).The GMV of the right insula,right postcentral gyms and right supramarginal grus were positively associated with B&P scores in patients with schizophrenia (P＜0.01,AlphaSim corrected),respectively.Conclusions These preliminary findings support that the aggression in schizophrenia is associated with GMV changes of brain regions in patients with schizophrenia.The right postcentral gyrus,the right insula and the right supramarginal gyrus may be involved in the neural mechanism of aggression in schizophrenia.

24 new cDNA sequences encoding cytochrome P450 were amplified respectively from deltamethrin susceptible and -resistant strains of the mosquito, Culex pipiens pallens, with a pair of degenerate primers according to the conservative amino acid sequences of CYP4 in insects by RT-PCR. Studies of molecular systematics show that the 24 new genes (alleles) belong to CYP4C, CYP4D, CYP4H and CYP4J subfamilies of the CYP4 family, and they were named by Cytochrome P450 Nomenclature Committee. Among the new genes (alleles), CYP4C23 may be a pseudogene, CYP4H13 has a retained intron 58 nucleotides in length, and CYP4J4V1 has a stop coden (TAG) in frame near the 3'-end.

Objective To learn the latest progress in the research of ω-3 unsaturated fatty acids and provide references for the related investigator through visualized analysis of the research of ω-3 unsaturated fatty acids published in our country.Methods China Biology Medicine disc was searched by computer from the beginning until December 31,2017.Bibliographic Item Co-occurrence Mining System (BICOMS) was used to extract and summarize the data of age,author,organization,province and key words and to produce a co-occurrence matrix.NetDraw of Ucinet 6.0 was employed to draw the social network diagram of the author,organization,province and key words.And cluster analysis of the key words was performed by gCLUTO 2.0.Results A total of 1 165 studies involviug 30 provinces and cities,854 research units,3 789 authors and 1 016 significative key words were included.The cooperation of author,organization and province needed to be further strengthened.The study focused on 3 aspects:the effects of unsaturated fatty acids on apoptosis/lipid in mice/rats with related genes of diabetes/cardiovascular disease/coronary artery disease;meta-analysis of immunonutrition to improve tumor/inflammation/sepsis;effect of fish oil fat emulsion on interleukin/C reactive protein in pneumonia/lung injury.Conclusions The research of ω-3 unsaturated fatty acids develops rapidly in China,but it is mainly concentrated in a few centers of cooperation agencies.There is little cooperation among different provinces,cities,areas,and organizations and the research topics needs to be further expanded.

Background/Aims: Roxadustat, an oral medication for treating renal anemia, is a hypoxia-inducible factor prolyl hydroxylase inhibitor used for regulating iron metabolism and promoting erythropoiesis. To investigate the efficacy and safety of roxadustat in patients undergoing peritoneal dialysis (PD) with erythropoietin hyporesponsiveness. Methods: Single-center, retrospective study, 81 PD patients (with erythropoietin hyporesponsiveness) were divided into the roxadustat group (n = 61) and erythropoiesis-stimulating agents (ESAs) group (n = 20). Hemoglobin (Hb), total cholesterol, intact parathyroid hormone (iPTH), brain natriuretic peptide (BNP), related indicators of cardiac function and high-sensitivity C-reactive protein (hs-CRP) were collected. Additionally, adverse events were also recorded. The follow-up period was 16 weeks. Results: The two groups exhibited similar baseline demographic and clinical characteristics. At baseline, the roxadustat group had a mean Hb level of 89.8 ± 18.9 g/L, while the ESAs group had a mean Hb level of 95.2 ± 16.0 g/L. By week 16, the Hb levels had increased to 118 ± 19.8 g/L (p < 0.05) in the roxadustat group and 101 ± 19.3 g/L (p > 0.05) in the ESAs group. The efficacy of roxadustat in improving anemia was not influenced by baseline levels of hs-CRP and iPTH. Cholesterol was decreased in the roxadustat group without statin use. An increase in left ventricular ejection fraction and stabilization of BNP were observed in the roxadustat group. Conclusions For PD patients with erythropoietin hyporesponsiveness, roxadustat can significantly improve renal anemia. The efficacy of roxadustat in improving renal anemia was not affected by baseline levels of hs-CRP0 and iPTH.

Objective:To explore the transcriptional regulation mechanism of transforming growth factor β 1 (TGF-β 1) on Meox1 and its effect on cell migration of adult human dermal fibroblasts (HDF-a). Methods:(1) HDF-a cells were cultured in RPMI 1640 complete medium (hereinafter referred to as routinely cultured). The cells were divided into TGF-β 1 stimulation group and blank control group. The cells in TGF-β 1 stimulation group were stimulated with 10 μL TGF-β 1 in the mass concentration of 1 mg/μL, while the cells in blank control group were stimulated with the equal volume of phosphate buffer solution. After 72 hours in culture, partial cells in both groups were collected for transcriptome sequencing. The genes with differential expression ratio greater than or equal to 2 and P<0.01 between the two groups were selected to perform enrichment analysis and analysis of metabolic pathways of the Kyoto Gene and Genome Encyclopedia with, and the expression value of Meox1 per million transcripts (TPM) was recorded ( n=3). Partial cells from the two groups were used to detect the Meox1 mRNA expression by real-time fluorescent quantitative reverse transcription polymerase chain reaction (RT-PCR) ( n=3). (2) Cultured HDF-a cells in the logarithmic growth phase (the same growth phase of cells below) were divided into empty plasmid group, Smad2 overexpression (OE) group, Smad3 OE group, and Smad4 OE group, which were transfected respectively with 2 μg empty pcDNA3.1 plasmid and pcDNA3.1 plasmids separately carrying Smad2, Smad3, and Smad4 for 6 hours, and then were routinely cultured for 48 hours. The Meox1 mRNA expression in the transfected cells of each group was detected by real-time fluorescent quantitative RT-PCR ( n=3). (3) HDF-a cells were routinely cultured and grouped the same as in experiment (1). After 72 hours in culture, the enrichment of Smad2, Smad3, and Smad4 protein on the Meox1 promoter in the cells of each group was detected by chromatin immunoprecipitation-quantitative PCR (ChIP-qPCR) ( n=3). (4) HDF-a cells were routinely cultured and divided into negative interference group, small interference RNA (siRNA)-Smad2 group, siRNA-Smad3 group, siRNA-Smad4 group, empty plasmid group, Smad2 OE group, Smad3 OE group, and Smad4 OE group, which were transfected respectively with 50 μmol/L random siRNA, siRNA-Smad2, siRNA-Smad3, siRNA-Smad4, 2 μg empty pcDNA3.1 plasmid and pcDNA3.1 plasmids separately carrying Smad2, Smad3, and Smad4 for 6 hours and then routinely cultured for 48 hours. The enrichment of Smad2, Smad3, and Smad4 protein on the Meox1 promoter in the cells of corresponding group was detected by ChIP-qPCR ( n=3). (5) Two batches of HDF-a cells were cultured and divided into negative interference group, siRNA-Meox1 group, empty plasmid group, and Meox1 OE group, which were transfected respectively with 50 μmol/L random siRNA, siRNA-Meox1, 2 μg empty pcDNA3.1 plasmid and pcDNA3.1 plasmid carrying Meox1 for 6 hours and then routinely cultured for 24 hours. One batch of cells were subjected to scratch test with the scratch width being observed 24 hours after scratching and compared with the initial width for scratch wound healing; the other batch of cells were subjected to Transwell assay, in which the migrated cells were counted after being routinely cultured for 24 hours ( n=3). (6) From January 2018 to June 2019, 3 hypertrophic scar patients (2 males and 1 female, aged 35-56 years) were admitted to the First Affiliated Hospital of Army Medical University (the Third Military Medical University) 8-12 months after burns. The scar tissue and normal skin tissue along the scar margin resected during surgery were taken, and immunohistochemical staining was performed to observe the distribution of Meox1 protein expression. Data were statistically analyzed with one-way analysis of variance and independent sample t test. Results:(1) After 72 hours in culture, a total of 843 genes were obviously differentially expressed between the two groups, being related to tissue repair, cell migration, inflammatory cell chemotaxis induction process and potential signaling pathways such as tumor necrosis factor, interleukin 17, extracellular matrix receptor. The TPM value of Meox1 in the cells of blank control group was 45.9±1.9, which was significantly lower than 163.1±29.5 of TGF-β 1 stimulation group ( t=6.88, P<0.01) with RNA-sequencing. After 72 hours in culture, the Meox1 mRNA expression levels in the cells of blank control group was 1.00±0.21, which was significantly lower than 11.00±3.61 of TGF-β 1 stimulation group ( t=4.79, P<0.01). (2) After 48 hours in culture, the Meox1 mRNA expression levels in the cells of Smad2 OE group, Smad3 OE group, and Smad4 OE group were 198.70±11.02, 35.47±4.30, 20.27±2.50, respectively, which were significantly higher than 1.03±0.19 of empty plasmid group ( t=31.07, 13.80, 13.12, P<0.01). (3) After 72 hours in culture, the enrichment of Smad2, Smad3, and Smad4 protein on the promoter of Meox1 in the cells of TGF-β 1 stimulation group was significantly higher than that of blank control group respectively ( t=12.99, 41.47, 29.10, P<0.01). (4) After 48 hours in culture, the enrichment of Smad2 protein on the promoter of Meox1 in the cells of negative interference group was (0.200 000±0.030 000)%, significantly higher than (0.000 770±0.000 013)% of siRNA-Smad2 group ( t=11.67, P<0.01); the enrichment of Smad2 protein on the promoter of Meox1 in the cells of empty plasmid group was (0.200 000±0.040 000)%, significantly lower than (0.700 000±0.090 000)% of Smad2 OE group ( t=8.85, P<0.01). The enrichment of Smad3 protein on the promoter of Meox1 in the cells of negative interference group was (0.500 0±0.041 3)%, significantly higher than (0.006 0±0.001 3)% of siRNA-Smad3 group ( t=17.79, P<0.01); the enrichment of Smad3 protein on the promoter of Meox1 in the cells of empty plasmid group was (0.470 0±0.080 0)%, which was significantly lower than (1.100 0±0.070 0)% of Smad3 OE group ( t=9.93, P<0.01). The enrichment of Smad4 protein on the promoter of Meox1 in the cells of negative interference group was similar to that of siRNA-Smad4 group ( t=2.11, P>0.05); the enrichment of Smad4 protein on the promoter of Meox1 in the cells of empty plasmid group was similar to that of Smad4 OE group ( t=0.60, P>0.05). (5) Twenty-four hours after scratching, the scratch healing width of cells in siRNA-Meox1 group was narrower than that of negative interference group, while that of Meox1 OE group was wider than that of empty plasmid group. After 24 hours in culture, the number of migration cells in negative interference group was significantly higher than that in siRNA-Meox1 group ( t=9.12, P<0.01), and that in empty plasmid group was significantly lower than that in Meox1 OE group ( t=8.99, P<0.01). (6) The expression of Meox1 protein in the scar tissue was significantly higher than that in normal skin of patients with hypertrophic scars. Conclusions:TGF-β 1 transcriptionally regulates Meox1 expression via Smad2/3 in HDF-a cells, thus promoting cell migration.