Table of Content

05 June 2021, Volume 41 Issue 6

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Original Articles

Establishment and evaluation of a two-hit model of mouse schizophrenia

YOU Peng-sheng, WEI Hui, XU Qi

2021, 41(6):  779-785. 

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Objective Prenatal infection and postpartum inflammatory experiences have been associated with increased risk of schizophrenia. To explore whether prenatal immune activation and prepubertal inflammation have cumulative impact on vulnerability to schizophrenia. Methods Polyinosinin-polycytidylic acid was injected into pregnant mice to establish maternal immune activation models on gestation day 9.5. Offspring were treated with poly I:C during puberty or adulthood as the second hit. 1)Using ELISA and quantitative real-time PCR to detect inflammatory response after administration of poly Ⅰ:C. 2)The effects of single or combined exposure to prenatal immune activation and prepubertal inflammation were examined by prepulse inhibition and locomotor reactions to dizocilpine(MK-801). Behavioral assessment took place when the animals reached adulthood. Results 1)Offspring from poly Ⅰ:C -treated mothers showed sensorimotor gating deficiency, as well as in the precipitation of behavioral hypersensitivity to the drugs dizocilpine, which were similar with schizophrenia. The phenotype of female was more pronounced than male. 2)However, double-hit model's sensorimotor gating deficient level was lower than that of prenatal immune activation model, and the phenotype of schizophrenia was relieved after second hit. Conclusions There is no synergistic pathological effect of prenatal immune activation plus pre-pubertal or adulthood inflammation.

Effect of NOTCH1 silencing mediated by spermine-pullulan combined with desloratadine on acute T lymphocyte leukemia cell line Jurkat

WANG Tian, XU Shi-lin, WANG Tao, WEN Tao, MENG Jie, LIU Jian, XU Hai-yan

2021, 41(6):  786-791. 

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Objective Using pullulan-spermine (Ps) as a carrier of NOTCH1 siRNA to find the effect of NOTCH1 silencing mediated by Ps combined with lysosomal penetration enhancer desloratadine (DL) on acute T lymphocytic leukemia cell line Jurkat. Methods A dynamic light scattering instrument was used to analyze the particle size and Zeta potential of siRNA/Ps complexes with different N/P ratios. Jurkat cells were incubated with siNOTCH1/Ps for 6 hours, then the medium was refreshed with RPMI 1640 complete medium containing 10 μmol/L DL and incubated for another 18 hours, real-time fluorescent quantitative PCR was used to detect the silencing efficiency of NOTCH1; flow cytometry was used to detect the cell cycle and apoptosis; Western blot was used to detect the expres- sion of C-MYC protein, which was the downstream of NOTCH1, and the expression of apoptosis-related proteins caspase 3 and cleaved caspase 3. Results The particle size of siRNA/Ps with N/P ratio of 5 was (203.97±1.07)nm, the zeta potential was (46.13±0.07)mV, and the particles were stable in 10% FBS medium. In Jurkat cells, compared with the no DL group, the siNOTCH1/Ps combined with 10 μmol/L DL induced cell cycle arrest in G₁ phase and promoted cell apoptosis (P＜0.05). At the same time, the expression level of cleaved caspase 3 protein in Jurkat cells increased, the expression level of C-MYC and caspase 3 protein decreased (P＜0.05). Conclusions Ps combined with lysosomal penetration enhancer DL effectively mediates the silencing of NOTCH1 in Jurkat cell, thereby inducing the G₁ phase arrest of acute T lymphocyte leukemia cells and promoting cell apoptosis.

STAT3 pathway is involved in interleukin-22 promoting expression of tight junction protein in human colorectal cancer cell line HT29

SONG Qian, HUANG Xue, QIN Meng-bin, ZHANG Jun-hong, JING Jie, LUO Yi-hua, LI Yu-xin

2021, 41(6):  792-798. 

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Objective To investigate the effect of interleukin-22(IL-22) on intestinal epithelial barrier tight junction and its mechanism. Methods Human colon cancer HT29 cells were divided into control group, IL-22 group(100 ng/mL), FLLL32 group(pretreatment with FLLL32 before IL-22 intervention) and colivelin group(pretreatment with colivelin before IL-22 intervention). The transfection treatment groups included: shNC group, shNC+ IL-22 group(infection with negative control virus before IL-22 intervention), shSTAT3 group(infection with LV-STAT3-RNAi virus), and shSTAT3+ IL-22 group(LV-STAT3-RNAi virus infection before IL-22 intervention). Western blot and real-time PCR were used to detect the protein and mRNA expression of STAT3, p-STAT3 tight junction protein ZO-1, occludin, claudin-1 and claudin-2 in HT29 cells. Transmission electron microscopy(TEM) was used to observe the changes in the structure of tight junctions of HT29 cells. Results The protein and mRNA expression level of STAT3, p-STAT3, ZO-1, occludin, claudin-1 and claudin-2 in IL-22 group were significantly higher than those in the control group(P＜0.05) with more compact, tight junction. Compared with IL-22 group, protein and mRNA expression level of the above indexes in FLLL32 group were significantly decreased(P＜0.05), the tight junction was poor, the expression of colivelin group was up-regulated(P＜0.05), the tight junction was intact. The shNC+ IL-22 group significantly increased the expression of all tightly bound proteins and mRNA compared with the shNC group(P＜0.05), the tight junction structure was more dense, while the shSTAT3+ IL-22 group showed no significant difference in protein and mRNA expression level when compared with theose of shSTAT3 group. Conclusions STAT3 signaling pathway is involved in IL-22 to promote the expression of tight junction proteins ZO-1, occludin, claudin-1 and claudin-2 in HT29 cells.

Macrophage J774A.1 exosome promotes the proliferation and migration of colorectal cancer cell lines MC38 and CT26

YANG Xu-dong, YU Hong-liang, LI Jing, ZHAO Chun-hua

2021, 41(6):  799-804. 

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Objective To investigate the effects of exosome secreted by macrophage J774A.1 on the proliferation and migration of mouse colorectal cancer cell lines MC38 and CT26 and potential mechanism. Methods The phenotype of J774A.1 cells was identified by flow cytometry.The morphological characteristics, size and surface markers of the exosome of J774A.1 cells were detected by transmission electronic microscopy, nanoparticle tracking analysis(NTA) and Western blot.The uptake of exosome by MC38 and CT26 cells was observed by fluorescence microscopy. Proliferation of MC38 and CT26 was detected by CCK8 assay. Migration ability of MC38 and CT26 was detected by wound healing assay. The expression of phosphorylated proteins p-MEK1/2 and p-ERK1/2 in proliferation-related pathways was detected by Western blot. Results J774A.1 cells are M2 macrophages expressing the cell surface marker molecules CD68 and CD206. J774A.1 exosomes can be taken up by colorectal cancer cells MC38 and CT26, and can promote their proliferation(P＜0.05), enhance their migration and activate MEK-ERK signaling pathway. Conclusions Macrophage J774A.1 exosomes can promote the proliferation and migration of colorectal cancer cell lines MC38 and CT26, which is potentially attributed to the increased phosphorylation of MEK-ERK signaling pathway.

Carboxyamidotriazole enhances the effects of mouse spleen lymphocytes on promoting the apoptosis of colon cancer cell lines MC38 and C26

GAO Hong-ting, HUANG Yu-qing, YANG Li-xing, MA Rui, WANG Yu-cheng, JU Rui, GUO Lei

2021, 41(6):  805-810. 

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Objective To investigate the direct effects of carboxyamidotriazole(CAI)on the survival and apoptosis of colon cancer cells(MC38 and C26)and its indirect effects through enhancing the anticancer effects of spleen lymphocytes in mice. Methods MC38 and C26 cells were cultured in vitro,spleen lymphocytes of mice were isolated,MC38 and C26 cells were co-cultured with spleen lymphocytes of mice,and the cells were treated with different concentrations of CAI. CCK-8 method was used to detect cell survival,flow cytometry was used to detect cell apoptosis and the proportion of CD4⁺CD3⁺ T cells and CD8⁺CD3⁺ T cells in spleen lymphocytes of mice. Results The number of living cells of MC38 and C26 cells decreased(P＜0.05,P＜0.01 and P＜0.001)and the apoptosis rate was increased(P＜0.01 and P＜0.001)after being treated with different concentrations of CAI. The number of living cells of MC38 and C26 cells was decreased(P＜0.001)upon being co-cultured with spleen lymphocytes of mice for 48 h, and the apoptosis rate was increased(P＜0.001). The number of living cells of MC38 and C26 cells in co- culture groups treated with different concentrations of CAI for 48 h further decreased(P＜0.001)and the apoptosis rate increased(P＜0.05 and P＜0.001)as compared with those co-cultured with spleen lymphocytes of mice without CAI. The proportion of CD4⁺CD3⁺ T cells and CD8⁺CD3⁺ T cells in spleen lymphocytes of mice was significantly increased(P＜0.05 and P＜0.01)after 48 h treatment with different concentrations of CAI. Conclusions CAI can inhibit the survival and promote apoptosis of MC38 and C26 cells,and enhance the effects of spleen lymphocytes in mice on inhibiting the survival and promoting apoptosis of MC38 and C26 cells through a potential mechanism of increasing the proportion of CD4⁺CD3⁺ T cells and CD8⁺CD3⁺ T cells in spleen lymphocytes of mice.

hnRNPA3 regulates hESCs pluripotency through modulating mRNA export

YANG Jia-bin, CHEN Zhong-yang, ZHOU Fan-qi, YU Jia, MA Yan-ni

2021, 41(6):  811-817. 

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Objective To explore the biological function of RNA binding protein heterogeneous nuclear ribonucleoprotein A3(hnRNPA3) in the maintenance of pluripotency of human embryonic stem cells(hESCs) and to reveal its new mechanism in RNA transport. Methods The interaction of hnRNPA3 with nuclear pore and localization were examined by immunoprecipitation of nuclear pore complex and cytoplasmic separation in hESCs. After inhibiting the endogenous expression of hnRNPA3, the expression of the pluripotency/differentiation marker genes was detected by qPCR, and the effect on the nucleoplasmic distribution of polyA⁺ RNA and OCT4 mRNA was tested by RNA fluorescence in situ hybridization to predict the binding of hnRNPA3 to known RNA export associated proteins. Results hnRNPA3 was localized in the nucleus and interacted with the nuclear pore. Inhibiting the endogenous expression of hnRNPA3 significantly inhibited the cloning-formation ability of hESC(P＜0.05) and resulted in the increase of the expression of cell differentiation markers(P＜0.001) and significantly inhibited the export of polyA⁺ RNA as well as OCT4 mRNA(P＜0.001). The protein interaction network predicted that hnRNPA3 may bind to about one-third of the known RNA export associated proteins. Conclusions hnRNPA3 regulates hESC pluripotency by impacting on mRNA export.

Effect of Dicer1 ablation on early neurodevelopment of mice

LIU Gao-ao, PENG Xiao-zhong, SHU Peng-cheng

2021, 41(6):  818-824. 

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Objective To explore the role of microRNAs(miRNAs) during early neurodevelopment in mammals with Dicer1 conditional knockout(cKO)mouse by RNA-seq. Methods The animals were divided into Ctrl group and cKO group. HE and immunofluorescence staining were used to observe the histological morphology; Extract total RNA of E14.5 D6-Cre-Dicer1 mice dorsal cortex, using RNA-seq technique to analyze differences at transcriptional level. Results Animals in cKO groups died within a month or so, abnormal development of cortical and hippocampal was recorded at P3. In cKO group transcription expression was different significantly,45 differentially expressed genes were screened, including 11 up-regulated and 34 down-regulated genes(P＜0.05)mainly enriched in Wnt signaling pathway, DNA damage response signaling pathway and others. Conclusions At E14.5 cortex, Dicer1 is involved in the regulation of multiple signaling pathways, affecting partial cortical laminar genes expres- sion, inhibiting the expression of lncRNA Xist and regulating cortical development.

Dynamic expression of RNA-binding proteins in spermatogenesis based on RNA-seq

LI Kai, MANG Xin-yu, ZOU Ding-feng, LI Meng-zhen, MIAO Shi-ying, WANG Lin-fang, SONG Wei

2021, 41(6):  825-830. 

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Objective To systematically characterize the dynamic expression pattern, stage specificity and co-expression pattern of RNA-binding protein (RBPs) and to predict their potential regulatory role in mouse spermatogenesis. Methods The whole transcriptome sequencing data of six spermatogenic cells types were integrated for analyzing the differentially expressed RBPs during spermatogenesis;STEM was used to analyze the dynamic expression pattern of RBPs; WGCNA was used to identify the RBPs co-expressed pattern; The differentially expressed and co-expressed RBPs were analyzed by the ClusterProfiler tool for GO function enrichment analysis. Results A total of 519 stage-specific RBPs were identified during spermatogenesis, and there were 7 dynamic expression patterns, of which RBPs at the meiotic stage accounted for the highest proportion; GO enrichment analysis showed that RBPs were mainly involved in the selective splicing, processing or translation of mRNA; WGCNA analysis showed that 246 RBPs were co-expressed, among which RBPs at the meiotic stage accounted for the highest proportion. Conclusions RBPs exhibit stage specificity and play a regulatory role in spermatogenesis with a coordinated expression mode. It functions mainly in the process of RNA processing or splicing in the early stage of spermatogenesis, and is have significant impact on the process of ribosome assembly or RNA translation in the later stage.

Identification and analysis of key genes in association with development and progression of adenocarcinoma at the gastroesophageal junction

WANG Yu-qian, CHEN Ya-mei, YANG Jie, LIN Yuan, LUO Ying-ying, ZHANG Shao-sen, WU Chen

2021, 41(6):  831-836. 

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Objective To explore the transcriptomic changes of adenocarcinoma at the gastroesophageal junction (ACGEJ) and identify key genes in the occurrence and progression. Methods RNA sequencing data of ACGEJ tumors and adjacent normal tissues samples from 58 patients who were recruited at the Linzhou Cancer Hospital and Linzhou Esophageal Cancer Hospital. The gene expression level of paired ACGEJ and adjacent normal tissue samples were compared to identify dysregulated genes. And the enriched pathways of these genes were analyzed by using gene set variation analysis (GSVA). LASSO regression model was used to test the association between the expression of dysregulated genes and the prognosis of ACGEJ patients, and the clinical features and prognosis associated gene set were applied to establish the nomogram model. Results Through transcriptomic profiling of tumor and non-tumor ACGEJ samples, 737 significantly differentially expression genes (DEGs) in ACGEJ were identified(｜log2FC｜＞1.2, Q＜0.05). The DEGs were related to tumor proliferation, growth, metastasis and metabolism. Among these DEGs, a total of 9 genes (ASF1B,ACTN1,KNL1,SAPCD2,TP53I11,DMBT1,CNFN,ID2,DPT) were identified to associate with the prognosis of ACGEJ patients by LASSO regression model. These 9 genes were further selected to build a prognostic model. The patients could be divided into two groups by this prognostic model, and the overall survival (OS) of the low-risk group was significantly higher than that of the high-risk group (P＜0.01). A nomogram was established which included the age, clinical stage and prognosis associated gene set for eventual clinical translation. Conclusions Several genes associated with the occurrence and progression of ACGEJ are identified. These genes may be potential treatment targets and prognostic biomarkers of this disease.

Relationship between metabolically healthy obesity and the prevalence of non-alcoholic fatty liver disease

XU Ke, HU Xiang, YU Wei-hui, YANG Li-juan, GU Xue-jiang

2021, 41(6):  837-841. 

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Objective To investigate the role of metabolically normal obesity in non-alcoholic fatty liver disease(NAFLD). Methods From 2011 to 2012, a cross-section survey with 4 757 participants enrolled from four communities in Wenzhou was conducted. Data collection was conducted through questionnaire survey, physical examination, laboratory examination, and multiple logistic regression analysis was used to comprehensively identify the risk factors of NAFLD. Results 1 068 individuals(22.4%) were diagnosed as metabolically healthy obesity. The prevalence of NAFLD was 43.5%. Among the various obesity phenotypes, the proportions ranked from high to low were metabolically unhealthy obese(MUO), metabolically unhealthy nonobese(MUNO), metabolically healthy obese(MHO) and metabolically healthy nonobese(MHNO). Obesity was defined by waist circumference and BMI, the risk of NAFLD in MHO population was 3.38 times(95% CI:2.13-5.37) and 4.69 times(95% CI:3.01-7.16) higher than that of MHNO, respectively. Conclusions MHO is associated with an increased risk of NAFLD in the Chinese population. It is necessary to screen for NAFLD in obese subjects regardless state of metabolic health.

Hedyotis diffusa inhibits the proliferation, migration and invasion of bladder cancer cell line KU-19-19 through up-regulating miR-485-5p

XU Lin-fei, ZHANG Hai-tao, LIU Sheng, CAI Hai-rong, HU En-ping

2021, 41(6):  842-847. 

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Objective To explore the effect and possible molecular mechanism of Hedyotis diffusa on the proliferation, migration and invasion of bladder cancer cells. Methods U-19-19 cells were divided into control group, Hedyotis diffusa group, miR-NC group, miR-485-5p group, anti-miR-NC group, anti-miR-485-5p group, Hedyotis diffusa +anti-miR-NC group, Hedyotis diffusa+anti-miR-485-5p group, Hedyotis diffusa+anti-miR-485-5p+LY294002 group. Cell counting kit (CCK-8), Transwell experiment, and real-time fluorescent quantitative PCR (RT-qPCR) were used to detect cell proliferation, migration, invasion and miR-485-5p expression. Western blot was performed to detect the phosphorylation levels of phosphatidylinositol-3-hydroxykinase (PI3K) and protein kinase B (AKT). Results After the intervention of Hedyotis diffusa, the proliferation rate, and the cell numbers of migrat and invade of KU-19-19 cells were significantly reduced, and the expression of miR-485-5p was significantly increased (P＜0.05). After overexpression of miR-485-5p, the proliferation rate, and the cell numbers of migrat and invade of KU-19-19 cells were significantly reduced (P＜0.05). After inhibiting the expression of miR-485-5p, the proliferation rate, and the cell numbers of migrat and invade of KU-19-19 cells were significantly increased (P＜0.05). Inhibition of miR-485-5p could reverse the inhibitory effect of Hedyotis diffusa on KU-19-19 cell proliferation, migration and invasion (P＜0.05). LY294002 could reverse the promoted effect of inhibiting miR-485-5p on proliferation, migration and invasion of KU-19-19 cells treated with Hedyotis diffusa (P＜0.05). Conclusions Hedyotis diffusa can inhibit the proliferation, migration and invasion of bladder cancer cells by up-regulating miR-485-5p. The mechanism may be related to the inhibition of PI3K/AKT signaling pathway.

Etomidate alleviates LPS-induced rat myocardial cell line H9C2 injury by upregulating miR-290-5p

LEI Jia-xiu, ZHU Ya-ping, ZHANG Yu

2021, 41(6):  848-853. 

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Objective To investigate the protective mechanism of etomidate (Eto) on lipopolysaccharide (LPS) induced myocardial injury. Methods H9C2 cells were divided into control (Co) group, LPS group (1 μg/mL LPS treatment for 6 h), Eto+LPS group, miR-con+LPS group, miR-290-5p+LPS group, Eto+anti-miR-con+LPS group, and Eto+anti-miR-290-5p+ LPS group. Tetramethylazozolium blue (MTT) method and flow cytometry were used to detect cell viability and apoptosis; real-time fluorescent quantitative PCR (RT-qPCR) was used to detect miR-290-5p expression. Enzyme-linked immunosorbent assay (ELISA) was used to detect tumor necrosis factor alpha (TNF-α) and interleukin 6 (IL-6) in cell culture fluid. Results Compared to the control group, the expression of miR-290-5p in H9C2 cells in the LPS group reduced (P＜0.05), the cell survival rate reduced(P＜0.05), the apoptosis rate increased (P＜0.05), and the level of TNF-α and IL-6 significantly increased (P＜0.05); compared with the LPS group, the expression of miR-290-5p in H9C2 cells in the Eto+LPS group was increased (P＜0.05), and the cell survival rate was decreased (P＜0.05), apoptotic rate was decreased (P＜0.05), TNF-α and IL-6 content were decreased (P＜0.05); compared with the miR-con+LPS group, the survival rate of H9C2 cells in the miR-290-5p+LPS group increased (P＜0.05), the apoptosis rate reduced (P＜0.05), the level of TNF-α and IL-6 reduced (P＜0.05); when compared with the Eto+anti-miR-con+LPS group, the survival rate of H9C2 cells in the Eto+anti-miR-290-5p+LPS group decreased(P＜0.05), and apoptosis rate increased (P＜0.05), the level of TNF-α and IL-6 increased(P＜0.05). Conclusions Etomidate reduces LPS-induced myocardial cell apoptosis and inflammatory response by up-regulating miR-290-5p.

Resveratrol inhibits kidney injury in renal transplantation rats

WANG Kai, LI Ming, QIAO Liang-wei, QU Qing-shan

2021, 41(6):  854-858. 

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Objective To explore the effect and mechanism of resveratrol on improving kidney injury in renal transplantation rats. Methods Forty rats were divided into sham group, model group (allogeneic kidney transplantation), high and low dose resveratrol groups [40 and 20 mg/(kg·d), intraperitoneal injection, for 7 consecutive days]. Twenty four hours after operation, the serum level of Cr, BUN, SOD and MDA was detected, the expression of Bax and Bcl-2 mRNA in renal tissues was tested by RT-qPCR, and the protein expression of IKKα, p-IκBα, and NF-κB p65 was detected by Western blot. Results Compared to the sham group, serum Cr, BUN and MDA level in the model group increased, SOD activity decreased, Bax mRNA expression in renal tissue increased, Bcl-2 mRNA expression decreased, and the expression of IKKα, p-IκBα, NF-κB p65 protein increased (P＜0.05); compared with the model group, resveratrol significantly reduced serum Cr, BUN and MDA content, while increased SOD activity, reduced Bax mRNA and IKKα, p-IκBα, NF-κB p65 protein expression, and increased Bcl-2 mRNA expression (P＜0.05). Conclusions Resveratrol has a protective effect on kidney injury in kidney transplantation rats, and the underlying mechanism is potentially related to the inhibition of activation of NF-κB signaling pathway.

Influence of DFG-STYK1/NOK on cell proliferation and G₁/S distribution in mouse fibroblast cell line NIH3T3

CHEN Jian-miao

2021, 41(6):  859-864. 

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Objective To investigate the effects of NOK added DFG motif on proliferation and G₁/S distribution of NIH3T3 cells. Methods SWISS MODEL homologous modeling was used to confirm the location of DFG addition; we used CCK-8 and MTT methods to measure the cell proliferation of NIH3T3 transiently transfected pcDNA3.0 or pcDNA3.0-DFG-NOK; Flow cytometry was used to measure cell cycle; Western-blot was used to exam the phosphorylation of Akt,Erk,STAT1,STAT3,JAK3 and the expression of cyclin D1. Results Compared with the pcDNA3.0 transient transfection: 1)The proliferation of pcDNA3.0-DFG-NOK over-expression NIH3T3 cells was inhibited. 2)The entrance of G₁/S in NIH3T3 cells transiently transfected pcDNA3.0-DFG-NOK was obstructed. 3)Transient transfection of pcDNA3.0-DFG-NOK could inhibit phosphorylation of Erk,STAT1,STAT3,JAK3 and expression of CyclinD1. Conclusions DFG-NOK inhibits proliferation of NIH3T3 cells and obstructs cell cycle G₁ phase entering into S phase.

Bioinformatic analysis of systemic lupus erythematosus based on miRNA microarray

KONG Jie, GAO Ying-ying, WANG Xue-qin

2021, 41(6):  865-870. 

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Objective To analyze the differential expressions of miRNA in monocyte derived dendritic cells (moDCs) from systemic lupus erythematosus (SLE) patients and healthy people by bioinformatic analysis. Methods The GSE79240 dataset was downloaded from the gene expression omnibus (GEO) database, differential miRNAs were screened by R language. Funrich software was used to analyze the gene ontology (GO) enrichment, Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment and transcription factors. MiRTarBase database was used to predict target genes. Results A total of 19 differential miRNAs were screened out,of which 11 were up-regulated and 8 were down-regulated.Biological process of GO enrichment was enriched in signal transduction and cell communication; cellular component of GO enrichment was enriched in nucleus and cytoplasm; molecular function of GO enrichment was expressed in transcription factor activity, ubiquitin-specific protease activity and receptor signalling complex scaffold activity. KEGG pathway was shown mainly in glypican signalling pathway, TRAIL signalling pathway and S1P signalling pathway. The transcription factors related to miRNA mainly included: SP1, SP4, EGR1 and POU2F1. Forteen miRNAs have been experimentally verified to have target genes. Conclusions The differential miRNAs in moDCs of SLE patients may play an important role in the pathogenesis of SLE and provide new clue for the diagnosis of SLE.

Effect of miR-26a on drug resistance of chronic lymphocytic leukemia cells by regulating PTEN gene

MAO Hong-bin, HE Ming

2021, 41(6):  871-875. 

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Objective To observe the regulation of microRNA-26a (miR-26a) on the drug resistance of chronic lymphocyte leukemia (CLL) cells through the phosphatase and tensin homolog deleted on chromosome ten (PTEN). Methods The experiment was divided into 4 groups: control group (L1210 cells), resistance group (L1210/DDP cells), NC group (transfected with empty plasmid+L1210/DDP cells), inhibitor group (transfected with miR-26a-inhibitor plasmid+L1210/DDP and inhibitor group (transfected with miR-26a-inhibitor plasmid+L1210/DDP cells). miR-26a and PTEN mRNA expression were detected by RT-qPCR. Dual luciferase experiment was used to detect the targeting of miR-26a to PTEN. MTT method was used to detect cell resistance to cisplatin (DDP). Flow cytometry was used to detect cell apoptosis rates. Western blot was used to detect of PTEN, phosphatidyl inositol-3-kinase (PI3K), serine-threonine protein kinase (AKT) protein expressions in cells. Results miR-26a inhibited the expression of PTEN in cells (P＜0.05). miR-26a targeted PTEN. Inhibiting the expression of miR-26a reduced the IC₅₀ value of cells to DDP, increased the rates of apoptosis, and up-regulated the expression of PI3K protein and p-AKT/AKT (P＜0.05). Conclusions Inhibiting the expression of miR-26a can reduces the effect of cell resistance through up-regulating the expression of PTEN.

Meteorological factors potentially cause spontaneous epistaxis in Xining area

DONG Chun-hua, ZHANG Ying, WANG Yi, MI Yu, ZHENG Hui-yuan, JIA Juan, SHI Kai-fang

2021, 41(6):  876-880. 

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Objective To identigy the relationship between the morbidity of epistaxis and the meteorological factors in Xining, Qinghai province. Methods The medical records of patients with spontaneous epistaxis from the department of otorhinolaryngology emergency and outpatient department from Jan 1, 2017 to Dec 31, 2017 were collected and retrospectively reviewed. Patients with a clear etiology for the epistaxis (traumatic, iatrogenic, tumor, foreign body,coagulopathic illness) were excluded. The monthly climatic data were collected from the Xining Meteorological Administration. The relationship between the incidence of epistaxis and meteorological factors were statistically analyzed with Pearson's correlation co-efficient. Results There were 1 219 cases of epistaxis in a year period. A significant negative correlation was found between temperature and epistaxis morbidity (P＜0.05). A significant negative correlation between humidity and epistaxis was also found(P＜0.05). Conclusions This is the first report about potential relationship beween meteorological factors and epistaxis in Xining. The spontaneous epistaxis is potentially attributed to meteorological factors in Xining.

Clinical Sciences

Clinical characteristics and treatment of 49 cases of testicular cancer

MEI Ru-qi

2021, 41(6):  881-884. 

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Objective To explore the clinical characteristics and treatment outcome of testicular cancer. Methods To review retrospectly the clinical data of 49 patients who were diagnosed with testicular cancer and treated with operation in the Urology Department of Peking University Cancer Hospital. Results Among the 49 patients, 11 had elevated preoperative alpha fetal protein(AFP) levels, all have the age as≤ 44 years; 21 had elevated β-human chorionic gonadotropin(β-hCG) level, including 19 cases with age as ≤ 44 years; 20 had elevated lactate dehydrogenase(LDH), including 18 cases with the age as ≤ 44 years. Two weeks after operation, there were 2 cases of abnormal AFP, both ≤ 44 years old; 6 cases of abnormal β-hCG, including 5 cases ≤ 44 years old; LDH were all normal. All patients underwent radical resection of testicular cancer, including 6 patients with retroperitoneal lymph node dissection(RLND), and 23 patients with postoperative radiotherapy/chemotherapy followed. Conclusions The significance of AFP, β-hCG and LDH is greater for the diagnosis and evaluation of patients < 45 years old. Markers with higher sensitivity and specificity are needed for evaluation. RLND can be an option for initial treatment.

Difference between uric acid stones and non-uric acid stones in conventional CT scanning

LIU Huan, YANG Xue-dong, SHI Feng-xiang, WU Zhe, SUN Hao, JIN Zheng-yu

2021, 41(6):  885-889. 

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Objective To investigate methods for predicting uric acid stones with conventional CT by measuring the CT value of stones, mean diameter size, density of stones, stone to kidney CT value ratio, and body width. Methods Patients underwent conventional CT scanning and were diagnosed postoperatively before uric acid stones and non-uric acid stones were detected by dual-energy CT.A total of 50 patients with uric acid stones and 60 patients with non-uric acid stones were enrolled, and after stones were detected by conventional CT scan, uric acid stones and non-uric acid stones were distinguished by dual-energy CT. The selected stones were ＞ 7 mm in long diameter. Data acquisition was performed on enrolled patients by a radiologist blinded to stone composition. The predictive value of each index was evaluated by independent sample t test, chi-square test, and receiver operating characteristic curve (ROC). Results There was great value in distinguishing uric acid stones from non-uric acid stones in stone CT value (P＜0.01), significant statistical difference in uniform stones or not and in size (P＜ 0.05). The CT scan showed a high predictive value for the diagnosis of uric acid stones, with AUC=0.948, sensitivity as 94%,and specificity as 85%.According to the ROC curve, the CT scaning value of 730 HU was adopted as the threshold for the diagnosis of uric acid stones, with a positive predictive value of 83.9%, a negative predictive value of 94.4%, and an accuracy of 80.1%. The size of the stone also had a predictive value, AUC=0.63, sensitivity 36%, and specificity 88.3%, and the mean diameter of the stone greater than 18.05 mm was defined as a diagnosis standard of uric acid stones according to the ROC curve, which had a positive predictive value of 72%, a negative predictive value of 62.4%, and an accuracy of 64.5%. Using a stone CT value of 730 HU as the threshold combined with a uniform stone density to predict uric acid stones, its specificity was 100% and the positive predictive value was 100%. Conclusions Conventional CT is of high value in differentiating uric acid calculi from non-uric acid calculi by measuring the mean diameter size and density of calculi. When evaluate CT result with or without homogenous stone density as the diagnostic criteria, the specificity and positive predictive value of diagnosing uric acid calculi are significantly higher.

Effects of celecoxib combined with IMRT on immune function and serum tumor markers in elderly patients with advanced non-small cell lung cancer

REN Meng-xian, LIANG Wei, LIU Yan

2021, 41(6):  890-894. 

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Objective To analyze the effect of intensity modulated radiation therapy IMRT(intensity modulated radiation therapy) combined with celecoxib on immune function and serum tumor markers in patients with non-small cell lung cancer(NSCLC). Methods Sixty-two patients with NSCLC in Taihe County People's Hospital from January 2017 to December 2019 were selected and randomly divided into two groups; thirty patients with radiotherapy alone were set as the control group, and thirty-two patients with IMRT combined with celecoxib were set as the experimental group. Fasting venous blood was collected before and 4 weeks after treatment to detect the serum tumor markers, including carbohydrate antigen 50(CA50), carbohydrate antigen 125(CA125), carcinoma-embryonic antigen(CEA), squamous cell carcinoma associated antigen(SccAg) and T lymphocyte subsets(CD3⁺, CD4⁺, CD8⁺) in the two groups. The short-term efficacy of the two groups was evaluated, and the survival curve was drawn by Kaplan Meier. Results The total effective rate of the experimental group was 54.55%, which was significantly higher than that of the control group(31.82%)(P＜0.05); after treatment, the serum CA50, CA125, CEA, SCCAg levels of the two groups were significantly lower than those before treatment(P＜0.05), and the experimental group was significantly lower than that of the control group(P＜0.05); after treatment, the test value of CD3⁺, CD4⁺ in the experimental group was significantly higher than those before treatment(P＜0.05), and the value of CD8⁺ was significantly higher than those before treatment. Counting of CD3⁺ and CD4⁺ in the control group was significantly lower than that before treatment(P＜0.05), but CD8⁺ counting was significantly higher than at before treatment(P＜0.05). There was significant difference in T lymphocyte subsets between the two groups(P＜0.05). The 3-year survival rate of the experimental group was 38.64%, which was significantly higher than that of the control group(18.18%)(P＜0.05). Conclusions Intensity modulated radiation therapy combined with celecoxib in the treatment of elderly patients with advanced NSCLC significantly reduces the level of serum tumor markers, effectively enhances the immune function, and significantly improves the treatment outcome as well as enhances survival rate.

Mini Reviews

Progress in the diagnosis and treatment of cognitive frailty

ZHAO Rui-xue, MA Ya-jun, LIU Hui, LI Xiao-dong, WANG Xiao-feng, JIANG Xiao-yan, LI Shu-juan

2021, 41(6):  895-898. 

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Cognitive frailty, which increases the risk of dementia, is a geriatric syndrome characterized by both physical frailty and cognitive impairment. It is associated with adverse health events such as functional disability, hospitalization, falls, all-cause mortality and the incidence of dementia, vascular dementia, neurocognitive disorders, with potential reversibility. A clear and unified operational definition of cognitive frailty in the future is of great significance for early recognition and intervention of cognitive frailty and secondary prevention of early or asymptomatic dementia.

Research progress on the role of epigenetic regulation in obstructive sleep apnea syndrome

CHEN Feng, LENG Yu-fang, REN Yi-xing, ZHANG Jian-min, LIU Xin, SHI Ya-jing

2021, 41(6):  899-903. 

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Chronic intermittent hypoxia plays an important role in the mechanism of various adverse reactions of obstructive sleep apnea syndrome (OSA). Epigenetic process participates in the occurrence and development of OSA and complications by affecting adaptive potential and phenotypic variation under chronic intermittent hypoxia. Therefore, understanding the effect of epigenetic regulation on OSA is helpful to provide new ideas for clinical phenotype and targeted therapy of OSA.

Research progress on novel molecules and new targets for the control of invasive fungi

GUO Xi, LAI Xin-xiu, YANG Mei-yan, LI Qiao-zhen, WANG Si-hang, SONG Zhang-yong

2021, 41(6):  904-908. 

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At present, the drug treatment for invasive fungal infection is challenged by serious adverse effects and drug resistance. Therefore, the research and development of safe and effective new antifungal drugs is a great importance to support the clinical treatment of invasive fungal diseases. This article reviewed new targets of invasive fungi, and explores the underlying mechanisms of fungal biofilm formation, cell wall,key metabolic enzymes, mitochondria in the aspects of small molecule inhibitors and secondary metabolites, which may provide reference for the research and development of new drugs.

Research progress on immune mechanism of exercise improving sarcopenia

DENG Yu-jie, LI Wei-xin

2021, 41(6):  909-913. 

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Sarcopenia occurred in aged population has become a research hotspot in recent years. Chronic low-level inflammation is an important factor leading to muscle loss. Exercise training is an effective countermeasure to antagonize skeletal muscle loss, and its mechanism is potentially explained by enhancing the response of immune system and Stimulating an in vivo anti-inflammatory response.The immune mechanisms which improves clinical outcome of sarcopenia by different exercise methods need more research for the prevention and treatment of sarcopenia in aged population.

Medical Education

Application value of computer clinical thinking training in clinical thinking teaching of rare diseases

CHEN Shi, LUO Yun-yun, BAI Xi, PAN Hui, ZHU Hui-juan

2021, 41(6):  914-917. 

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Computer clinical thinking training is a teaching method that uses the computer technology to simulate real clinical scenarios. Compared with the traditional clinical thinking training mode, it has its unique advantages. This article introduces the current status of computer clinical thinking training as well as its application values in rare diseases teaching, existing problems and solutions. And we emphasize its potential in clinical thinking teaching. This new method guaranteeing the homogeneity of physician training will advance the progress of medical teaching reformation.

Evaluation of hands-on instruction versus video demonstration for vacuum delivery training

ZHONG Yi-feng, SONG Ying-na, TANG Ping-ping, YIN Jie

2021, 41(6):  918-922. 

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Objective To evaluate two methods applied to simulation training of vacuum delivery. Methods A total of 49 residents were randomly divided into two groups. One group was given on-site instruction by directors and hands-on demonstration (referred to as “tutor group”) and the other group was given demonstration through video (referred to as “video group”). Before and after the training, the two groups were surveyed with a self-confidence questionnaire. Written examination was implemented and compared results of group trained by vedio demonstration and the group of on-site instruction by tutors. The practical operation was evaluated by the Objective Structured Assessment of Technical Skills (OSATS). Results The on-site instructed group had a significantly higher score than that of the video group (4.96±0.20 vs 4.68±0.48, P＜0.05). The score of written examination of on-site instructed group was significantly higher than that of the video group (83.00±7.18 vs 70.56±10.37, P＜0.001). In the OSATS, the score of equipment integrity in on-site instrcted group was significantly higher (0.96±0.20 vs 0.52±0.51 P＜0.01), the correct placement of the flexion point (0.50±0.51 vs 0.08±0.28, P＜0.01) and the Objective total score (6.79±1.02 vs 5.32±0.95, P＜0.001) of on-site instructed group were all The same phenomenon was also found in the soft tissue clamping (0.63±0.50 vs 0.32±0.48, P＜0.05). Conclusions The on-site and tutor instructed simulation training has shown a better outcome comparing to the video demonstration . However,both groups can enhance the students'confidence in learning and using operative vaginal delivery skills.

Investigation of training mode and curriculum construction of medical leadership

JIA Ying-hua, BAI Xi, HE Feng-yu, PAN Hui

2021, 41(6):  923-926. 

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Objective To investigate the current situation of medical students' cognition and cultivation of medical students on medical leadership, providing the basis for the leadership cultivation and curriculum construction in medical colleges. Methods Taking medical students of Peking Union Medical College and one Chinese Medical College as the research objects, 256 valid data were collected through the network questionnaire, and the survey contents was conducted analysis statistically with descriptive study. Results The analysis result of survey data suggest that there is little difference between clinical and non-clinical students in the cognition of the importance of medical leadership. Peking Union Medical College achieved significant results in cultivation of medical leadership on the exploration and practice. In terms of medical leadership training and curriculum construction, it is suggested to adopt the form of practice-based or case study. Meanwhile, communication ability and the ability of organization and coordination are the abilities that need to be improved mostly. Conclusions The independent opinion and communication skill training for leadership in Peking Union Medical College should be extensively adopted by other medical schools. It is necessary to continuesly strengthen the training of medical leadership and construction of education courses focusing on the capacity building for communication, organization and team spirit and other core skills of medical leadership.

Necessity and evaluation of teaching for medical postgraduates in the view point of knowledge needed as physician-scientists

WANG Yi, YANG Ai-rong, YANG Yong-li, YU Zhang-bin, ZHANG Ying, MA Xin-chun

2021, 41(6):  927-930. 

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Objective To evaluate the necessity and effect of knowledge needed for research in the education of research-oriented medical graduates. Methods In March 2021, medical experts introduced the knowledge needed for the first-year medical graduates in Qinghai University Affiliated Hospital. Questionnaire survey was conducted to understand the current situation of graduates' understanding of the aim of training for the physician-scientists before the training and evaluate its necessity. The learning outcomes were evaluated by the comparison before and after the training. Results Only 55.16% of graduates understood the difference between traditional clinicians and physician-scientists before training, and 54.37% knew the difference between traditional clinical reasoning and clinical research logics, which indicated that it was very necessary to carry out teaching of knowledge as physician-scientists to medical graduates. After training, 95.78% of graduate students knew the difference between traditional clinicians and physician-scientists, and 86.75% of them knew the difference between traditional clinical reasoning and physician-scientists logics, which was significantly higher than that before training (P＜0.01). Conclusions Medical graduates have insufficient knowledge needed by physician-scientists. Training can significantly improve their understanding of physician-scientists.

Investigation and analysis of the current teaching situation of young physicians in internal medicine department at Peking Union Medical College Hospital(PUMCH)

SHI Wen, QU Xuan, SHI Xiao-chun, LI Hang, ZHANG Feng-chun

2021, 41(6):  931-934. 

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Objective To investigate the current situation, motivation, difficulties and needs of young physicians participating in teaching activities in internal medicine department at PUMCH. Methods We surveyed young attendings and residents who worked at internal medicine wards of Peking Union Medical College Hospital from November 2018 to October 2019 about their current tasks, motivation, difficulties and needs in terms of teaching. Results Based on visual Analog Scale, the mean score of attendings'interest in teaching is 8.0±1.8, and teaching work accounted for 25.3%±14.1% of all working hours. The most important motivation for teaching is self-improvement via teaching. The biggest difficulty faced by attendings is high pressure on clinical and research work (72.4%, 42/58). The most voted need is to increase the impact of teaching on promotion (48.3%, 28/58). The mean score of residents'interest in teaching is 7.5±1.8,and teaching work accounted for 24.8%±15.2% of all working hours. The most important motivation for teaching is self-improvement via teaching. The biggest difficulties faced by residents include high pressure on clinical and research work (50.6%, 41/81) and lack of teaching skills (25.9%, 21/81), etc. The most voted need is teaching skill trainings (42%, 34/81). Conclusions Young physicians are highly motivated to teaching with self-improvement as the main driving force. Measures should be taken to support young physicians to better balance clinical, research and teaching, and to improve their teaching results.