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BACKGROUND:Previous studies have successfully constructed erythropoietin-overexpressed umbilical cord mesenchymal stem cells.It was found that the apoptosis of ischemic and hypoxic human neuroblastoma cell line(SH-SY5Y)was significantly reduced by erythropoietin-overexpressed umbilical cord mesenchymal stem cells. OBJECTIVE:To explore the possible neuroprotective mechanisms of erythropoietin-overexpressed umbilical cord mesenchymal stem cells against ischemic-hypoxic SH-SY5Y and their associated epigenetic mechanisms. METHODS:Oxygen-glucose deprivation was applied to ischemia-hypoxia-induced SH-SY5Y cell injury,and multifactorial assays were applied to detect the expression levels of inflammatory factors in the cells before and after hypoxia and co-culture,respectively,with mesenchymal stem cells,as well as lentiviral-transfected null-loaded plasmids of the negative control mesenchymal stem cells and erythropoietin-overexpressed umbilical cord mesenchymal stem cells.The expression levels of supernatant inflammatory factors were detected by multifactor assay after co-culture.Proteomics was used to detect the differentially expressed proteins of negative control mesenchymal stem cells and erythropoietin-overexpressed umbilical cord mesenchymal stem cells.Cleavage under targets and tagmentation sequencing was applied to detect genomic H3K4me2 modification,and joint analysis was conducted with RNA-sequencing.Lentiviral vector infection was applied to construct the stable knockdown of REST in SH-SY5Y cells.qRT-PCR and western blot assay were performed to detect the expression level of REST.The apoptosis was detected by flow cytometry after co-culture of oxygen-glucose deprivation treatment with erythropoietin-overexpressed umbilical cord mesenchymal stem cells.The expression difference of H3K36me3 group proteins was detected by western blot assay,and transcriptome sequencing was performed to analyze the differentially expressed genes. RESULTS AND CONCLUSION:(1)Compared with the control group,monocyte chemotactic protein 1,interleukin-6,interleukin-18,and interleukin-1 beta,interferon α2,and interleukin-23 levels significantly increased in the cerebrospinal fluid supernatant of patients with ischemic-hypoxic encephalopathy(P<0.01).(2)After co-culturing SH-SY5Y cells with erythropoietin-overexpressed umbilical cord mesenchymal stem cells under ischemia and hypoxia,the expression levels of monocyte chemotactic protein 1 and interleukin-6 were significantly reduced.(3)Analysis of protein network interactions revealed significant downregulation of monocyte chemotactic protein 1,interleukin-6 related regulatory proteins CXCL1 and BGN.(4)Transcriptome sequencing analysis found that pro-inflammatory genes were down-regulated,and functional enrichment of histone modifications,and the expression of transcription factors REST and TET3 significantly up-regulated in the erythropoietin-overexpressed umbilical cord mesenchymal stem cell group compared with the negative control mesenchymal stem cell group.(5)Combined analysis of transcriptome sequencing and cleavage under targets and tagmentation revealed changes in epigenetic levels as well as significant activation of the promoter regions of transcription factors REST and TET3.(6)Stable knockdown REST in SH-SY5Y cells was successfully constructed;the transcript levels of REST mRNA and protein expression were both decreased.(7)After the REST knockdown SH-SY5Y cells were co-cultured with erythropoietin-overexpressed umbilical cord mesenchymal stem cells,apoptosis was significantly increased and H3K36me3 expression was significantly decreased.Transcriptome sequencing results showed that the expression of inflammation-related genes Aldh1l2 and Cth,as well as apoptosis-suppressor genes Mapk8ip1 and Sod2 was reduced at mRNA transcription level(P<0.01).(8)It is concluded that erythropoietin-overexpressed umbilical cord mesenchymal stem cells activated the expression of REST and TET3 by altering the kurtosis of H3K4me2 and upregulated the modification level of H3K36me3,which in turn regulated the expression of inflammation-related genes Aldh1l2 and Cth,as well as apoptosis-suppressor genes Mapk8ip1 and Sod2,and facilitated neuronal survival.
ABSTRACT
Objective The nursing treatment ability scale of patients with nuclear radiation damagein the hospital was developed to provide an evaluation basis for improving the nursing ability of nurses with nuclear radiation damage. Methods The scale was prepared by literature review, expert interview and expert consultation, and a total of 330 clinical nurses from a third-class hospital was randomly selected as the research objects. The scales were issued for item analysis and reliability and validity test. Results The scales were divided into 6 dimensions, including basic knowledge of nuclear radiation damage, specialized equipment use ability, specialized ward management ability, basic nursing ability, specialized nursing ability and self-ability recognition, with 51 items. After exploratory factor analysis, there were 6 principal components, and the cumulative interpreted variance was 70.757%. The χ2, df, χ2/df, CFI, IFI, TLI, NFI, PNFI, PCFI, RMSEA fitting indexes of confirmatory factor analysis were all acceptable. Cronbach's α coefficient was 0.976, the retest reliability was 0.823, and the S-CVI (S-CVI/UA) was 0.84. The evaluation content validityS-CVI (S-CVI/AVE) was 0.98, and the content validity I-CVI of the item level was 0.78～1.00. Conclusion The items and dimension Settings of this scale have been tested, and all indicators met the requirements. The reliability and validity test results were good. It can be used as a scale for preliminary evaluation of hospital nursing ability of patients with nuclear radiation damage.
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Objective:To explore the correlation between serum C1q, thyroid hormone (serum FT 3, FT 4, TSH) and depression by detecting the difference of serum C1q, thyroid hormone (serum FT 3, FT 4, TSH) between depression patients and normal people. Methods:A total of 275 depressive patients(depression group) and 275 healthy controls(healthy group) were recruited.The serum levels of C1q, FT 3, FT 4 and TSH were compared between the two groups.The serum levels of C1q, FT 3, FT 4 and TSH in depression patients with different age, gender, course of disease and HAMD were compared.Further regression analysis was used to evaluate the effects of serum C1q, FT 3, FT 4 and TSH levels on the incidence and severity of depression in patients with depression. Results:Serum C1q, FT 3, TSH of depression group(C1q: 203(165, 239)mg/L, FT 3: (4.39±0.70)pmol/L, TSH: 1.69(1.17, 2.46)mIU/L) were significantly lower than those of healthy group(C1q: (236.25±27.06)mg/L, FT 3: 4.61(4.29, 4.95)pmol/L, TSH: 2.04(1.42, 3.01)mIU/L)(all P<0.01). Compared with the serum indexes in depression group, the level of C1q and TSH in men(C1q : 188.00 (164.00, 221.00) mg/L, TSH: 1.52(1.13, 2.16)mIU/L) were significantly lower than those in women(C1q : 213.00 (168.25, 247.75) mg/L, TSH: 1.85(1.28, 2.57)mIU/L)( P<0.05), and the level of FT 3 and FT 4 in men(FT 3: 4.64 (4.23, 5.06) pmol/ L, FT 4: 16.76(15.05, 18.20)pmol/L) were significantly higher than those in women (FT 3: 4.34 (3.82, 4.72) pmol/L, FT 4: 15.92(14.35, 17.40)pmol/L). Serum C1q ( B=-0.020, P<0.01, OR95% CI: 0.980 (0.975, 0.985)), FT 3 ( B=-0.576, P<0.01, OR95% CI: 0.562 (0.408, 0.775)), TSH ( B=-0.274, P<0.01, OR95% CI: 0.761 (0.648, 0.893)) level were the influencing factors of depression. Conclusion:Serum C1q and thyroid hormone may be involved in the pathogenesis of depression, and may be affected by gender factors.
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By reverse transcription-polymerase chain reaction (RT-PCR), an open reading frame of pathogenesis-related protein 1 (PR1) was isolated from Panax notoginseng and named as PnPR1. Molecular and bioinformatic analyses of PnPR1 revealed that an open reading frame of 501 bp was predicted to encode a 166-amino acid protein with a deduced molecular mass of 18.1 kD. Homology analysis showed that the deduced amino acid sequence of PR1 protein of Panax notoginseng had a high similarity with other higher plants had the same conservative structure domain of cysteine-rich secretory protein (CAP). The recombinant expressed plasmid pET28a(+)-PnPR1 was expressed in Escherichia coli BL21. The expression conditions were optimized by induction at different times, different temperatures, different IPTG concentrations and different giving times. The optimum expression condition was 0.4 mmol.L-1 IPTG at 28 degrees C for 20 h. The successful expression of PnPR1 provides some basis for protein purification and preparation of the monoclonal antibody.
ABSTRACT
Stepping up clinical consultation control is an effective way to improve the level of patient care and medical quality. A retrospective analysis of relevant clinical data led to the discovery that among the leading causes of clinical misdiagnosis and mistreatment were limitation in the scope of knowledge on the part of specialized physicians, inadequate collection of medical record data, over-reliance on accessory examinations, and one-sided clinical thinking. It is pointed out that stepping up clinical consultation control is of great importance as it can help solve clinical difficulties and raise the rate of accuracy in diagnosis; promote professional exchange and raise the rate of success in emergency treatment; cultivate and enhance young doctors expertise in diagnosis and treatment; protect the interests of patients and reduce medical disputes. The paper also puts forward measures for improving and implementing the system of clinical consultation: revising consultation criteria, standardizing consultation procedure, strengthening consultation supervision, and setting up a system of consultation result follow-up and retrospective case discussion.