Animals ; Neural Stem Cells ; metabolism ; Rats ; Rats, Wistar ; Semaphorins ; metabolism

To generate recombinant adenovirus expression vector of human Sema4C gene and observe its expression in mouse myoblasts cell line C2C12 for ensuring easy access to investigate the role of Sema4C gene during myogenesis. The recombinant plasmid was packaged and amplified after being transfected in HEK293 cells through Lipofectamine. After infecting C2C12 myoblasts with recombinant adenovirus vector, the adenoviral infection efficiency was determined by confocal microscope which showed that the expression of green fluorescence could be detected at 12h and then reached peak at 24h after recombinant adenovirus infection. The infection efficiency was almost 100% confirmed by FACS examination. Detection of WB indicated that the expression of Sema4C in C2C12 of recombinant adenoviral infection group was significantly higher than that of the control group (P

Our previous study demonstrates that hypoxia promotes human bone marrow-derived mesenchymal stem cell (hMSC) proliferation. The aim of the present study was to investigate the gene profile involved in this process by using cDNA microarray. Cultured hMSCs were treated with hypoxia (3% O(2)) for 4 h, 12 h, 24 h, 36 h, 48 h and 72 h, respectively. Then these cells were collected to prepare total RNA. Hypoxia-induced gene expression profile was examined and analyzed by GenePix Pro 4.0 software. Some of cDNA microarray results were confirmed by RT-PCR. Microarray analysis identified that 282 genes expressed differentially, of which most were involved in metabolism. The number of differentially expressed genes at different hypoxia time points was different, and most genes were regulated after 24-hour hypoxia. Among the 282 differentially expressed genes, 4 hypoxia-inducible factor 1 (HIF-1) targeted genes and 10 genes that changed at 3 continuous time points were found. The results obtained indicated that 4 HIF-1 targeted genes, i.e., transforming growth factor beta3 (TGFbeta3), phospho-glycerate kinase 1 (PGK1), insulin-like growth factor binding protein 3 (IGFBP3) and BCL2/adenovirus E1B 19 kDa interacting protein 3 (BNIP3), displayed up-regulated pattern at 36 h under hypoxia. BNIP3 displayed a dynamically up-regulated pattern at 12, 36 and 72 h under hypoxia. However, TGFbeta3 and PGK1 were down-regulated at 72 h. In addition, the gene expressions of adenylate kinase 3-like 1 (HAC), neurofilament light polypeptide 68 kDa (NEFL), N-myc downstream regultated gene 1 (NDRG1), discoidin domain receptor family member 1 (DDR1), tribbles homolog 3 (TRIB3), nucleoprotein (AHNAK) and eukaryotic elongation factor selenocyteine-tRNA-specific (EESTS) were up-regulated. Moreover, the gene expressions of EESTS, NEFL were up-regulated at 5 different time points under hypoxia. Furthermore, it was found that the gene expressions of histone cluster 1 (HIS1) and transferring receptor (TFRC) were down-regulated. These results suggest that the proliferation of hMSCs induced by hypoxia is a complex process in which a number of genes may be involved.
Bone Marrow Cells ; cytology ; Cell Hypoxia ; Cell Proliferation ; Cells, Cultured ; Gene Expression Profiling ; Gene Expression Regulation ; Humans ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Oligonucleotide Array Sequence Analysis ; Oxygen ; metabolism ; Transcriptome

Objective: To investigate the differences in clinical characteristics, diagnosis, and treatment of pediatric septic shock in pediatric intensive care unit (PICU) among hospitals of different levels. Methods: This retrospective study enrolled 368 children with septic shock treated in the PICU of Beijing Children's Hospital, Henan Children's Hospital, and Baoding Children's Hospital from January 2018 to December 2021. Their clinical data were collected, including the general information, location of onset (community or hospital-acquired), severity, pathogen positivity, consistence of guideline (the rate of standard attainment at 6 h after resuscitation and the rate of anti-infective drug administration within 1 h after diagnosis), treatment, and in-hospital mortality. The 3 hospitals were national, provincial, and municipal, respectively. Furthermore, the patients were divided into the tumor group and the non-tumor group, and into the in-hospital referral group and the outpatient or emergency admission group. Chi-square test and Mann-Whitney U test were used to analyze the data. Results: The 368 patients aged 32 (11, 98) months, of whom 223 were males and 145 females. There were 215, 107, and 46 patients with septic shock, with males of 141, 51, and 31 cases, from the national, provincial, and municipal hospitals, respectively. The difference in pediatric risk of mortality Ⅲ (PRISM Ⅲ) scores among the national,provincial and municipal group was statistically significant (26(19, 32) vs.19(12, 26) vs. 12(6, 19), Z=60.25,P<0.001). The difference in community acquired septic shock among the national,provincial and municipal group was statistically significant (31.6%(68/215) vs. 84.1%(90/107) vs. 91.3%(42/46), χ²=108.26,P<0.001). There were no significant differences in compliance with guidelines among the 3 groups (P>0.05). The main bacteria detected in the national group were Klebsiella pneumoniae (15.4% (12/78)) and Staphylococcus aureus (15.4% (12/78)); in the provincial group were Staphylococcus aureus (19.0% (12/63)) and Pseudomonas aeruginosa (12.7% (8/63)), and in the municipal group were Streptococcus pneumoniae (40.0% (10/25)) and Enteric bacilli (16.0% (4/25)). The difference in the proportion of virus and the proportion of 3 or more initial antimicrobials used among the national,provincial and municipal group was statistically significant (27.7% (43/155) vs. 14.9% (13/87) vs. 9.1% (3/33), 22.8%(49/215) vs. 11.2%(12/107) vs. 6.5%(3/46), χ²=8.82, 10.99, both P<0.05). There was no difference in the in-hospital mortality among the 3 groups (P>0.05). Regarding the subgroups of tumor and non-tumor, the national group had higher PRISM Ⅲ (31(24, 38) vs. 22 (21, 28) vs.16 (9, 22), 24 (18, 30) vs. 17(8, 24) vs. 10 (5, 16), Z=30.34, 10.45, both P<0.001), and it was the same for the subgroups of in-hospital referral and out-patient or emergency admission (29 (21, 39) vs. 23 (17, 30) vs. 15 (10, 29), 23 (17, 29) vs. 18 (10, 24) vs. 11 (5, 16), Z=20.33, 14.25, both P<0.001) as compared to the provincial and municipal group. There was no significant difference in the in-hospital mortality among the 2 pairs of subgroups (all P>0.05). Conclusion: There are differences in the severity, location of onset, pathogen composition, and initial antibiotics of pediatric septic shock in children's hospitals of different levels, but no differences in compliance with guidelines and in-hospital survival rate.
Female ; Male ; Humans ; Child ; Retrospective Studies ; Shock, Septic/therapy* ; Hospitalization ; Intensive Care Units, Pediatric ; Hospitals, Pediatric