Table of Content

05 January 2020, Volume 40 Issue 1

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Original Articles

CX3CR1 mediates neuronal apoptosis by regulating Ca²⁺ influx to affect the prognosis of ischemic stroke mice

LIU Chang, ZHANG Shen, ZHOU Fei-hui, XIA Yang, LI Zhi-gao, WANG Jin-kun, TANG Zhi-wei

2020, 40(1):  1-8. 

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Objective To observe the changes of chemokine receptor 1(CX3CR1) expression in neurons after cerebral ischemia, and to explore its function mechanism. Methods Wild-type C57/BL6 mice and CX3CR1 gene knockout mice were set up as control group (sham operation group) and permanent middle cerebral artery occlusion(pMACO) model, respectively. Cerebral ischemia range and infarction area were detected after 30min by MRI, and apoptosis was detected by immunofluorescence triple staining.Western blot was used to detect the expression of CX3CR1 protein.In vitro, primary neurons were cultured to establish an oxysaccharide dissection (OGD) cell ischemia model. Cell survival rate was detected by MTT colorimetry, expression of CX3CR1 on neurons was detected by immunofluorescence, and the changes of Ca²⁺ concentration in neurons was observed by laser confocal microscopy. Results Compared with the control group, the expression of CX3CR1 in affected side of pMACO significantly increased in wild-type mice (P＜0.05), and CX3CR1 was co-expressed with the apoptotic protein caspase-3 in neurons.Compared with wild-type ones, the ischemic injury area of CX3CR1 gene knockout mice was similar 30 min after pMACO, but the infarct area of CX3CR1 gene knockout mice 24 h after pMACO was smaller than that of the control group (P＜0.05).In vitro, the expression of CX3CR1 significantly increased after OGD of primary neurons (P＜0.05). Knockout of CX3CR1 on neurons can reduce the excitatory injury mediated by glutamate and significantly increase the cell survival rate. Meanwhile, the knockout of CX3CR1 reduces the speed and quantity of glutamate-mediated Ca²⁺ flowing into neurons. Conclusions Ischemia can induce the expression of CX3CR1 on neurons, and CX3CR1 on neurons mediates the apoptosis of neurons by regulating Ca²⁺ internal flow.

Effect of miR-29a on proliferation and apoptosis of fibroblast-like synoviocyte in rheumatoid arthritis

GUO Xiong-fei, WANG Ting, TANG Li-xin

2020, 40(1):  9-15. 

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Objective To investigate the effect of miR-29a on the activity and apoptosis of fibroblast-like synoviocyte(FLS) in rheumatoid arthritis(RA). Methods The normal human FLS (n-FLS) and RA patients FLS (RA-FLS) were isolated and cultured,and the expression level of CHI3L1 protein was detected by Western blot. miR-29a mimics, si-NFAT5 and miR- 29a+NFAT5 were transfected into RA-FLS for 48 h. Then,the expression levels of miR-29a and NFAT5 mRNA were detected by RT-qPCR, the cell viability and apoptosis were detected by MTT assay and flow cytometry,respectively and the expression of p38, p-p38, cleaved-caspase3 proteins were detected by Western blot. Targeting relationship between miR-29a and NFAT5 was validated by dual fluorescent reporter gene detection system. Results Compared with n-FLS, the expression level of CHI3L1 protein in RA-FLS was increased significantly (P＜0.05). After over-expression of miR-29a or inhibition of NFAT5, the cell viability decreased, the apoptosis rate increased, the expression of p-p38 decreased, and the expression of cleaved-caspase3 increased significantly(P＜0.05). Luciferase activity of RA-FLS was significantly decreased in miR-29a and wild-type NFAT5 3′UTR co-transfection group, but increased in anti-miR-29a and wild-type NFAT5 3′UTR co-transfection group (P＜0.05). Over-expression of miR-29a significantly decreased the expression of NFAT5, and inhibition of miR-29a significantly increased the expression of NFAT5 (P＜0.05).Over-expression of NFAT5 can weaken the effect of over-expression of miR-29a on RA-FLS activity and apoptosis (P＜0.05). Conclusions miR-29a can inhibit the activity of RA-FLS and induce its apoptosis by targeting NFAT5 to down-regulate p38 MAPK signaling pathway.

RNA-seq analysis of the transcriptome of the cerebral cortex in the db/db mice

GUO Bao-lu, BAI Tao, GUO Jie, LIU Meng-meng, YANG Cai-hong, ZHANG Yi, FAN Yan-ying

2020, 40(1):  16-23. 

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Objective To investigate the dysregulated genes, pathways and networks involved in type 2 diabetes-derived brain dysfunctions via performing whole transcriptome sequencing on the cerebral cortex of db/db and wild-type (WT) mice. Methods Male db/db and WT mice were divided into two groups (n=9). The body weight and blood glucose were measured at week 8 and 24. The 24-week-old animals were sacrificed for cerebral cortex collection. RNA-sequencing and analysis of differentially expressed genes (DEGs), GO functional enrichment, KEGG pathway enrichment and PPI network were performed. Results A total of 306 differential transcripts, including 178 up-regulated and 128 down-regulated ones were identified in the db/db mice as compared to WT ones. Among all differentially expressed genes, 43 genes (such as Clcnka and Trim17) were found to be up-regulated and 59 genes (such as Arih1 and Nectin-3) were down-regulated. The protein-protein interaction network was constructed with thirteen nodes as hub genes. All the nodes were down-regulated genes and majorly belonged to mitochondrially encoded families. The GO functional enrichment showed changes in various categories as compared to WT. KEGG enrichment analysis revealed that target genes were mainly enriched in metabolic pathways, Parkinson's disease, Alzheimer's disease, and the involved DEGs mainly regulate oxidative phosphorylation process. Conclusions The relationship between type 2 diabetes and central nervous system impairments, and the potentially related genes, pathways and networks are identified.

Nicotine promotes apoptosis of rat cardiomyocyte cell line H9C2 cultured with high glucose/high fat

LIU Wen-xia, LIU Cai-hong, GUO Rui, MENG Zhi-jun, GAO Jia, WANG Ya-jing

2020, 40(1):  24-29. 

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Objective To study the apoptosis and mechanism of nicotine(Nic) in H9C2 cells under high glucose and high fat(HGHL) conditions. Methods H9C2 cells were cultured with different concentrations of nicotine (6×10^-8 mol/L Nic, 6×10^-7 mol/L Nic, 6×10^-6 mol/L Nic) for 2 hours, and HGHL (33 mmol/L glucose and 500 μmol/L palmitate) was added for 24 hours, cell proliferation was detected by CCK8 assag, cell viability was measured by lactate dehydrogenase (LDH) kit.The expression of Bax, Bcl-2, cleaved-caspase3 and caspase3 and the phosphorylation levels of ERK1/2, JNK and p38MAPK were detected by Western blot. The supernatant ROS level was determined by flow cytometry. Results Compared with normal controls, high glucose, high fat and nicotine combined with high glucose and high fat significantly inhibited cell viability and increased LDH level in the supernatant(P＜0.001). The apoptosis rate of HGHL and Nic+HGHL cells was significantly increased (P＜0.001), and the expression of cleaved-caspase3 protein was significantly increased (P＜0.01). The expression of Bcl-2/ Bax protein was significantly decreased (P＜0.01). ROS level was most significantly expressed in the Nic+HGHL group (P＜0.001). At the same time, HGHL activated MAPK phosphorylation level. ERK1/2 phosphorylation level was significantly increased in Nic+HGHL group (P＜0.01). ROS scavenger (NAC) significantly reduced caspase3 protein expression (P＜0.001). Conclusions Nicotine, high glucose and high fat can cause H9C2 cell damage and increase apoptosis, which is closely related to the regulation of ROS and MAPK signaling pathway.

LncRNA HEIH regulates proliferation and apoptosis of lung cancer cells through miR-98-5p

JIANG Qing-feng, YU Yong-kui, CHENG Jin-hua, SHEN Si-ning, XING Wen-qun

2020, 40(1):  30-36. 

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Objective To investigate the effect of lncRNA HEIH on proliferation and apoptosis of lung cancer cells and its mechanism. Methods Human lung epithelial cells BEAS-2B and lung cancer cell lines A549, A427, H1299 and TKB-1 were cultured, and the expression level of HEIH was detected by RT-qPCR. Si-HEIH and miR-98-5p mimics were transfected into A549 cells inorder to silence the expression of HEIH in A549 cells or over-expressing miR-98-5p. Then cell proliferation was detected by MTT assay, apoptosis was examined by flow cytometry, and the levels of CCND1, caspase-3, SHH, GLI-1, PTCH and SUFU proteins were measured by Western blot. The dual luciferase reporter gene assay verified the relationship between HEIH and miR-98-5p. Results Compared with the normal lung epithelial cells BEAS-2B, the levels of HEIH in lung cancer cell lines,A549, A427, H1299 and TKB-1 were significantly increased (P＜0.05). Among them,the level of HEIH in lung cancer celllines A549 was the highest. Therefore, A549 cells were selected as the research object in the subsequent experiments. Silencing HEIH expression or over-expression of miR-98-5p reduced the A value of A549 cells after 12, 48 or 72 hours(P＜0.05)(MTT assay), increased apoptosis rates (P＜0.05) and inhibited CCND1 protein expression(P＜0.05), and promoted caspase-3 protein expression (P＜0.05). Over-expression of miR-98-5p also inhibited the mRNA and protein expression of SHH and GLI-1 in A549 cells (P＜0.05), and promoted the mRNA and protein expression of PTCH and SUFU (P＜0.05). Over-expression of HEIH reversed the effect of over-expression of miR-98-5p on proliferation and apoptosis of A549 cells and mRNA and protein expression of SHH, GLI-1, PTCH and SUFU. Conclusions Silencing HEIH expression may inhibit the proliferation of lung cancer cells and promote apoptosis by targeting miR-98-5p through Hedgehog signaling pathway.

Sulfhydrated Ras related protein 7a inhibits HBV replication in human hepatocellular carcinoma cell line HepG2.2.15

SONG Jia-ru, ZHU Xi-lin, WU Xiao-pan, LIU Ying

2020, 40(1):  37-40. 

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Objective To investigate the mechanism of sulfhydrated Ras related protein 7a (Rab7a) in HepG2.2.15 cells and its effect on hepatitis B virus (HBV) replication. Methods The sulfhydrated Rab7a and sulfhydrated sites of Rab7a were verified by biotin switch method. The effect of the sulfhydrated Rab7a on the level of HBV replication markers was detected by enzyme-linked immunosorbent assay, RT-qPCR and Western blot. Results In HepG2.2.15 cells, H₂S enhanced the sulfhydration level of endogenous Rab7a (P＜0.01). NaHS (H₂S rapid release donor) enhanced the expression of sulfhydration of exogenous Rab7a, but the mutation of sulfhydration site was not affected by NaHS (P＜0.01). In HepG2.2.15 cells, H₂S enhanced the sulfhydrylation level of endogenous and exogenous Rab7a; the sulfhydrylated Rab7a inhibited the expression of HBsAg, HBeAg and HBV-DNA in supernatant, as well as HBV-cccDNA, 3.5RNA, total RNA and HBcAg protein in HepG2.2.15 cells (P＜0.05). Conclusions Sulfhydrated Rab7a inhibites the replication of HBV in HBV positive cell line HepG2.2.15.

Comparison of biological characteristics of CD146 positive subpopulations of mesenchymal stem cells derived from three different sources

WANG Qiao-ling, ZHANG Xiao-tong, XU Yan, ZHANG Nai-yue, ZHU Rong-jia, ZHAO Chun-hua

2020, 40(1):  41-47. 

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Objective To characterize the biological features of CD146⁺ cells from umbilical cord mesenchymal stem cells(UCMSCs), adipose-derived mesenchymal stem cells(AMSCs) and bone marrow mesenchymal stem cells(BM-MSCs). Methods MSCs from three tissues were sorted by magnetically activated cell sorting(MACS) to obtain highly purified CD146⁺ subpopulations. The phenotypes and ultra-structures of these three MSC were analyzed by flow-cytometry and transmission electron microscopy. The differentiation abilities were tested by adipogenic and osteogenic inductions. The expression of genes relative to adipogenesis, osteogenesis, stemness or angiogenesis were determined by RT-qPCR. Results CD146⁺ cells derived from three kinds of tissues had similar ultra-structures and the ability of angiogenesis. CD146⁺ AMSCs expressed highest levels of the OCT-4, SOX2 and NANOG genes.Under the same induction conditions, CD146⁺ AMSCs showed higher potential for adipogenesis and CD146⁺BM-MSCs showed higher potential for osteogenesis.However, CD146⁺ UMSCs had little ability for adipogenesis and osteogenesis. All of CD146⁺ MSCs from 3 tissues expressed BFGF, VEGF, Ang-1 and EGF although CD146⁺ BM-MSCs had stronger ability of angiogenesis in vitro. Conclusions CD146⁺ MSCs derived from the 3 sources exhibit different biological characteristics.These results provide a useful biological basis for further studies relative to application of MSCs.

Curcumin inhibits TGF-β1-induced phenotypic transition of human kidney tubular cells (HKCs)

CHEN Min-jia, DU Juan, YUAN Dan-feng, ZHANG Shu, HUANG Hong, ZHU Fang-qiang

2020, 40(1):  48-53. 

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Objective To study the effects of curcumine on renal tubular epithelial-mesenchymal transition (EMT) and its possible mechanisms. Methods The effect of curcumin on the morphology of human kidney cells (HKCs) in the presence of TGF-β1 alone or in combination with different concentration curcumine was observed by phase contrast microscopy. The expressions of E-cardhrin, keratin, vimentin, alpha smooth muscle actin (α-SMA),fibroblast-specific protein 1 (FSP1) and key proteins of AKT/mTOR pathway were analyzed by immunohistochemical stain and Western blot. Results HKC cells showed a classic cobblestone morphology. Exposure of HKCs to TGF-β1 for 72 h induced a complete conversion of the epithelial cell to myofibroblast. When HKCs were co-incubated with TGF-β1 and various concentrations of curcumin for 72 h, curcumine maintained the epithelial morphology in a dose-dependent manner (from 12.5 to 50 μg/mL), antagonizing TGF-β1-induced the down-regulation expression of E-cardhrin, keratin and the up-rgulation of FSP1, vimentin and α-SMA. Importantly. Curcumine effectively suppressed the activity of the Akt/mTOR pathway in HKCs as demonstrated by a remarkable reduction of the Akt, mTOR, p70S6K, 4E-BP1和eIF4E phosphorylation. Conclusions Curcumine is a potent inhibitor of TGF-β1-induced epithelial-mesenchymal transition (EMT) and antagonizes TGF-β1-driven fibrogenesis through inhibition of the activity of the Akt/mTOR pathway.

Expression and purification of human perforin by sf9 insect cells

CHEN Feng-ye, LIU Meng-yu, ZHOU Ya-bo, LIANG Xiao-yu, HUANG Bo, XIE Jing

2020, 40(1):  54-59. 

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Objective To optimize the purification method of human perforin. Methods Perforin with His tag was expressed by sf9 insect cells, and then protein affinity chromatography was performed by Ni column device. The physicochemical property and biological activity of the purified protein was analyzed by coomassie brilliant blue staining, silver staining and flow cytometry. Results This method was more convenient and economical than traditional protein purification methods like ion exchange chromatography, molecular sieve and hydrophobic interaction chromatography, which providing new ideas for further research of perforin and purification of other protein. Conclusions Perforin can be expressed by sf9 insect cells and is purified by a simple method with biological activity.

Screening and identification of the microRNA targeting c-SKI

WANG Juan, LI Peng, LYU Zhong-ying, ZHANG Ying, CAO Gui-qiu

2020, 40(1):  60-66. 

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Objective To screen and validate the microRNA targeting c-SKI and associated with fibrosis. Methods Bioinformatics methods was used to predict the candidate miRNAs targeting c-SKI, RT-qPCR was used to detect the expression of candidate miRNAs and c-SKI gene in myocardial fibroblasts, and the most significant miRNAs (miR-155a-5p/miR-17a-5p) were selected. Building SKI-3 ′-UTR wild type (c-SKI-wt) and mutant (c-SKI-mut) plasmid,miR-155a-5p/miR-17a-5p mimics and mimics NC,miR-155a-5p/miR-17a-5p inhibitor and inhibitor NC were co-transfected into the 293T cells. The c-SKI-wt or c-SKI-mut. Dual-luciferase reporter assay was performed to detect the luciferase activity in different groups.miR-155a-5p,miR-17a-5p mimics and inhibitor were transfected to myocardial fibroblasts.Western blot was used to detect the expression of c-SKI in each group. Results 1)after the primary screening miR-155a-5p and miR-17a-5p showed the most obvious inhibition on c-SKI(P＜0.01). 2)Compared with the NC group, the c-SKI reporter gene luciferase activity significantly decreased inmiR-155a-5p/ miR-17a-5p mimics group (P＜0.05),and the c-SKI reporter gene luciferase activity was obviously upregulated in miR-155a-5p and miR-17a-5p inhibitor group (P＜0.05).3)Compared with the NC group, the expression of c-SKI significantly down-regulated in miR-155a-5p and miR-17a-5p mimics group, but up-regulated in miR-155a-5p and miR-17a-5p inhibitor group as detected by Western blot(P＜0.01). Conclusions miR-155a-5p and miR-17a-5p can target c-SKI, and regulate the expression of c-SKI in myocardial fibroblasts.

Effects of IL-33 on HaCAT cell line and psoriasis-like lesion in mouse model

DUAN Ya-ju, WANG Wen-qian, HU Hua, FU Dan-dan, SONG Xiang-feng, TIAN Zhong-wei

2020, 40(1):  67-72. 

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Objective To investigate the effects of interleukin-33(IL-33)on HaCAT cells and psoriasis-like inflammation induced by imiquimod(IMQ) in mice. Methods Twenty-four hours after HaCAT cells were plated, the cell proliferation and autophagy-related protein were detect by CCK8 and Western blot. A psoriasis-like lesion model was established in mouse. Thirty female BALB/c mice of clean grade were randomly divided into three groups: control group, IMQ group, and IL-33 treatment group. After 6 days of continuous treatment, the lesions on the back of the mouse were observed. Results The treatment of IL-33 could enhance the proliferation of HaCAT cells and promote the expression of LC3, Beclin1 and p-STAT3 proteins. In the mouse psoriasis-like model, the thickened ear skin, scaly erythema and erythematous lesions on the back were seen and thickening of the epidermis, epidermal extension with parakeratosis were shown. These phenomena were more obvious in IL-33 treatment group(P＜0.05). The expression of autophagy-related proteins in the skin lesions of the IL-33 treatment group was significantly decreased compared with the control group and IMQ group(P＜0.05). Conclusions IL-33 promotes the proliferation of HaCAT cells, induces autophagy, and aggravates the occurrence and development of psoriatic lesions induced by imiquimod in mice, which may be involved in the activation of the STAT3 signaling pathway.

Pantoprazole induces bradyarrhythmia in mice with hypokalemia

LI Shuai, ZHU Wen-gen, YAN Xia, LAI Wei, YU Jian-hua, WAN Rong, HONG Kui

2020, 40(1):  73-77. 

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Objective To investigate the mechanism of arrhythmia induced by pantoprazole (PPZ) in mice with hypokalemia. Methods Mice were randomly divided into control group, PPZ group [20 mg/(kg·d), intraperitoneal injection for 5 weeks], furosemide (FS) group and FS+PPZ group. After 5 weeks, the ECG parameters (heart rate, PR interval, QRS interval, QTc interval) and spontaneous arrhythmia were monitored. The changes of the mRNA and protein expression of hyperpolarization-activated and cyclic nucleotide-cation channels 2 and 4 (HCN2 and HCN4), cardiac voltage-gated sodium channel alpha subunit (SCN5A), L-type calcium channel alpha subunit (CACNA1C) and T-type calcium channel alpha subunit (CACNA1G) were analysed by real-time fluorescence quantitative PCR and Western blot technology. Results Compared with control group, the HR was significantly decreased and PR interval was prolonged in PPZ group (P＜0.01, P＜0.05). Compared with FS group,the HR and PR interval in FS+PPZ group were further decreased and prolonged (P＜0.01, P＜0.05), and 4 mice developed sinus arrest (SA) (P＜0.05). The expressions of HCN4 mRNA and protein in PPZ group were lower than those in control group (P＜0.01), respectively, and the expression in FS+PPZ group was lower than those in FS group (P＜0.01). The expression of HCN2 mRNA in PPZ group was lower than that in control group(P＜0.05), which was further decreased in FS+PPZ group compared with FS group (P＜0.01). Conclusions It is found that pantoprazole may lead to bradyarrhythmia in mice under hypokalemia, which may be related to abnormal expression of HCN2 and HCN4.

Estradiol valerate improves endometrial receptivity induced by clomiphene in rat with polycystic ovary syndrome

SHAO Rui-feng, HE Chi-hua, WANG Ying

2020, 40(1):  78-82. 

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Objective To investigate the effect of estradiol valerate (E₂V) on endometrial receptivity induced by clomiphene in polycystic ovary syndrome (PCOS) rats. Methods Rats were divided into control group (administered 0.9% sodium chloride solution), and PCOS model group (administered 50 mg/kg clomiphene), E₂V low, medium and high-dose intervention group [administered (0.2, 0.5, 1.0)mg/kg E₂V+50 mg/kg clomiphene]. Serum sex hormone level was compared. Morphological change of endometrium was examined by ELISA. The expression of estrogen receptor (ER), progesterone receptor (PR), homeobox gene (Hox)A10, empty ventilator homeobox (Emx)2, integrin α_vβ₃ mRNA and protein in the endometrium were detected by real-time fluorescence quantitative polymerase chain reaction (RT-qPCR) and Western blot. Results Compared with the control group, luteinizing hormone/follicle-forming hormone (LH/FSH), testosterone (T), relative expressions of Emx2 mRNA and protein were significantly increased (P＜0.05), while estradiol (E2) level, glandular area, glandular area, endometrial thickness and glandular/interstitial area ratio, relative expressions of ER, PR, HoxA10, integrin α_vβ₃ mRNA and protein were significantly decreased (P＜0.05). Compared with PCOS model group, the changes of above indexes in E₂V low, medium and high-dose intervention group were opposite (P＜0.05). Conclusions E₂V can improve endometrial receptivity of PCOS clomiphene-induced ovulation rats. E₂V is potentially related to the up-regulation of ER, PR, HoxA10, integrinα_vβ₃ and down-regulation of Emx2.

Neferine inhibits Aβ1-42-induced PC12 cell injury through regulating the expression of miR-29a-3p/AQP4

ZENG Li-min, LU Zhao-hui, ZHOU Li-ping, ZHU Chao-xia

2020, 40(1):  83-91. 

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Objective To explore the role and molecular mechanism of Neferine (Nef) on PC12 cells injury induced by Aβ1-42. Methods PC12 cells were divided into control group, model group(4 μg/mL Aβ1-42 cultured for 24 h) and low, medium and high-dose neferine intervention group. The PC12 cell viability was determined by MTT method and cell apoptosis was detected by flow cytometry. Protein expression of aquaporin4 (AQP4), Bcl-2 and Bax was analyzed by Western Blot, and miR-29a-3p, AQP4 mRNA were detected by qPCR. Biological information prediction and dual luciferase gene report analyzed the targeting relationship between miR-29a-3p and AQP4. miR-29a-3p, si-AQP4 were transfected into PC12 cells induced by Aβ1-42, or anti-miR-29a-3p was transfected into cells and performed high-dose Nef intervention to investigate the effects on cell survival and apoptosisin PC12 cells injury induced by Aβ1-42. Results Compared with the model group, the cell viability rate, Bcl-2 and miR-29a-3p expression of neferine group were significantly increased, and the cell apoptosis rate, Bax, AQP4 mRNA and protein of neferine group expression were significantly decreased (P＜0.05). AQP4 was the target gene of miR-29a-3p. Both over-expression of miR-29a-3p and inhibition of AQP4 expression significantly increased the viability rate of PC12 cells injury induced by Aβ1-42 and the expression level of Bcl-2, and decreased the apoptosis rate and Bax protein level. Inhibitiion of expression of miR-29a-3p reversed the promotion effect of Neferine on the survival of PC12 cell injury induced by Aβ1-42 and the level of Bcl-2 protein, as well as the inhibition effect of Neferine on the apoptosis rate and Bax protein expression. Conclusions Neferine inhibits PC12 cell injury induced by Aβ1-42, promotes cell survival and inhibits apoptosis by miR-29a-3p/AQP4.

Pseudolaric acid B promotes apoptosis of human laryngeal carcinoma cell line HEp-2

YANG Meng-sheng, LI Li, LIU Xue-feng, YANG Xiao-long

2020, 40(1):  92-96. 

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Objective To investigate the effects of pseudolaric acid B (PAB) on the proliferation and apoptosis of HEp-2 cells. Methods Cell grouping and treatment: cells were treated with 0, 0.5, 1, 2, 4, 8 and 16 μmol/L PAB for 24, 48 and 72 h respectively; MTT assay was used to detect cell viability; flow cytometry was used to detect apoptosis; The expression of Bcl-2, Bax and caspase-3 protein and mRNA was detected by Western blot and qPCR. The expression of PARP and c-PARP was detected by immunohistochemistry. Results PAB inhibited HEp-2 proliferation in a time-concentration dependent manner. Compared with the control group, the apoptosis rate of HEp-2 cells in 4 μmol/L and 8 μmol/L PAB groups was significantly increased(P＜0.01). Compared with the control group, the expression of Bcl-2 and PARP in HEp-2 cells of 4 μmol/L and 8 μmol/L PAB groups were significantly decreased(P＜0.05), and the expression of Bax, caspase-3 and c-PARP was significantly increased(P＜0.05). Conclusions PAB can activate the mitochondrial apoptosis pathway thus induce apoptosis of HEp-2 cells and inhibit cell proliferation.

Propofol inhibits invasion and migration of esophageal squamous carcinoma cell line KYSE150 through miR-218

GAO Hong, JIN Hui, LIU Chun-zhi, YANG Jing-yuan

2020, 40(1):  97-104. 

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Objective To investigate the effect of propofol on invasion and migration of esophageal squamous cancer cell line KYSE150 and potential mechanism. Methods KYSE150 cells were treated with 0, 2.5, 5 and 10 μg/L propofol,Transwell chamber and scratch assay were used to detect the invasion and migration of cells after 24 h. RT-qPCR was used to detect the expression of miR-218 in cells. Western blot analysis was performed to detect the expression of HMGB1 protein. Bioinformatics software prediction and dual luciferase reporter gene assay were performed to detect the targeting relationship between miR-218 and HMGB1, Western blot to detect the regulation of miR-218 on HMGB1 protein. After transfection of miR-218 inhibitor or pcDNA3.1-HMGB1-GFP over-expression vector plasmid was constructed by liposome method, the expression of miR-218 was detected by RT-qPCR, the expression of HMGB1 protein was detected by Western blot, Transwell chamber and scratch test. The effect of down-regulating miR-218 or up-regulating HMGB1 expression on invasion and migration of 10 μg/L propofol-treated KYSE150 cells was examined. Results Propofol inhibited the invasion, migration and expression of HMGB1 protein in KYSE150 cells in a concentration-dependent manner and promoted the expression of miR-218. Dual luciferase reporter gene experiments confirmed that HMGB1 was a target gene of miR-218, and Western blot analysis confirmed that miR-218 negatively regulated the expression of HMGB1 protein. Down-regulation of miR-218 expression reversed the inhibitory effect of propofol on invasion and migration of KYSE150 cells. Up-regulation of HMGB1 expression reversed the inhibitory effect of propofol on invasion and migration of KYSE150 cells. Conclusions miR-218 plays an active role in the inhibition of invasion and migration of esophageal squamous cell carcinoma KYSE150 cells by propofol, and its mechanism is potentially related to targeted regulation of HMGB1.

Clinical Sciences

Clinical significance of testing blood parameters on the diagnosis of common microcytic anemia patients

ZHANG Ruo-xi, LI Hong-min, WU Jie, HAN Bing

2020, 40(1):  105-109. 

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Objective To evaluate the clinical significance of different hematology parameters on microcytic anemia patients. Methods Blood samples from patients in PUMCH with different microcytic anemia from October 2017 to September 2018 were tested for routine blood tests, cellular hemoglobin of reticulocytes (CHr), serum ferritin (SF), soluble transferrin receptor (sTfR), hepcidin and other iron metabolism parameters. Data from different diseases were analyzed and compared. Results Twenty-six iron deficiency anemia (IDA), 12 anemia of chronic disease (ACD), 12 thalassemia (TA), 12 refractory anemia with ring sideroblast (RARS) and 17 normal controls (NC) were tested. The level of MCV (mean corpuscular volume) in IDA and TA group were lower than that of control(P＜0.01). MCV level in TA group was even lower than that in IDA. The concentrations of mean corpuscular hemoglobin concentration(MCHC) in ACD, IDA, TA, RARS groups were lower than that in NC (P＜0.01), among which IDA had the lowest level. All types of anemia, except RARS, had higher level of reticulocyte counts than NC(P＜0.01), among them, TA group had the highest level. The level of CHr in ACD, IDA and TA groups was lower than that of NC(P＜0.01). SF concentration in IDA group was significantly lower than NC (P＜0.01), and was also lower than other 3 groups. ACD and RARS had higher concentrations of SF than NC(P＜0.01). As for the levels of transferrin saturation (TS), ACD, IDA, and TA groups were lower than NC (P＜0.01), while RARS was significantly higher than NC(P＜0.01) and also higher than the other 3 types of anemia. The level of sTfR and sTfR index(sTfR/log SF) in IDA group was significant higher than that in NC(P＜0.01), also higher than ACD, TA and RARS groups.ROC curve showed that SF and sTfR index facilitated differentiating IDA from other types of microcytic anemia. Conclusions Routine tests and iron metabolism related parameters from the peripheral blood tests may support the preliminary identification of different kinds of microcytic anemia and further differential diagnosis.

Analysis of publications on patient safety: a bibliometric study based on Citespace

GAO Xiao-xing, CHEN Zheng, YANG Ying-ying, PAN Hui

2020, 40(1):  110-114. 

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Objective To summarize the research trend with numerous publications on patient safety during the past 10 years and find out recent hot spot in this field. Methods Four thousands publications for the past 10 years focusing on patient safety were selected in the citation database of Web of Science. Bibliometric and visualization software named Citespace was utilized for co-existence analysis and burstness detection in analyzing key words and cited references. Results Organizations distributed in developed countries such as the United States, the United Kingdom accounts for the majority of publications. The most frequent key words in the research field of patient safety were care, adverse event, health care, error, quality etc. Newly emerged burstness turned to attitude question naire, experience, knowledge and patient safety culture etc. Conclusions A wide and in-depth series of studies in patient safety has been carried out from 2009 to 2019 especially in developed countries with a hot spot on patient safety culture. More studies from Chinese researchers on patient safety are warranted to form China's own experience.

Mini Reviews

Research progress in the regulation of bone homeostasis by long noncoding RNAs

FAN Lin-yuan, LI Hong-ling

2020, 40(1):  115-119. 

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Bone formation and bone resorption are two important processes of bone homeostasis maintenance, and the imbalance of bone homeostasis can lead to a variety of bone metabolic diseases. Long noncoding RNAs (lncRNAs) are a subset of RNAs with only rarely encoding proteins and play important regulatory role in regulation of gene expression, life development and disease occurrence.In bone metabolic diseases, lots of lncRNAs are abnormally expressed. LncRNAs regulate bone formation process through regulating chondrogenic differentiation, osteogenic differentiation and bonematrix mineralization. By affecting the differentiation and maturation of osteoclasts,lncRNAs are involved in the regulation of bone resorption.

Medical Education

Application of duty competency evaluation system in standardized training of internal medicine residents

HUANG Can, ZHAO Xue, LI Xiao-qing, SHI Xiao-chun, LI Hang, YAN Xiao-wei, ZHANG Feng-chun

2020, 40(1):  120-123. 

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Objective To establish a duty competency evaluation system in Peking Union Medical College Hospital (PUMCH) which can stratify residents based on their performance and authenticate the ability to practice medicine independently on duty. Methods Residents enrolled from PUMCH Internal Medicine Training program in 2017 and 2018 were evaluated for duty competency, the number of test and time needed to pass the test were analyzed. Results The numbers of tests needed to pass were not statistically significant(χ²=9.12,P＞0.05)in postdoctoral residents, undergraduate residents and residents from Beijing training base. Time needed to pass exam showed difference(χ²=96.27,P＜0.05) in the three resident groups. The results showed that this exam helped to stratify residents. Most residents passed exam after 6 months' internship and was competent for independ practice on duty. Conclusions In standardized training of residents, duty competency evaluation system can help to assess their capability on duty, and stratified training will help residents fulfill improvement and assure clinical safety. Residents from Beijing training base needed more clinical teaching based on their performance on the exam. Most residents passed the exam after training.

Application of blending teaching in biochemistry course for undergraduates

YAN Jing, HU Bo, JIANG Jing-yi, WANG Guo-li, FENG Chen, WANG Hua-qin

2020, 40(1):  124-127. 

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Objective To develope blending teaching method for inspiring students' interest and autonomy in learning, and improving students' learning outcome. Methods The clinical case guidance method, the combination of PBL (problem based learning) and LBL (lecture based learning), “rain classroom” and mind map summing up method were used in biochemistry lectures. Results Students were able to gradually establish the thinking mode of construction of meaning and integrate the complex knowledge to build up their own knowledge system. Blending teaching method made classroom teaching diversified and enriched to improve students' learning autonomy, which switched the thinking mode from the passive memory model to the active thinking mode of meaning construction. Conclusions The blending teaching method can improve students' learning interest, arouse their attention to the basic medical courses and build solid foundation for clinical medical courses learning.

Investigation and analysis of communication status and training needs between ICU nurses and patients' families

LI Zun-zhu, LI Zhen, ZHANG Yuan-yuan, LIU Jin-bang, LUO Hong-bo, HE Huai-wu, LONG Yun

2020, 40(1):  128-131. 

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Objective To investigate the learning needs of communication training between Intensive Care Unit(ICU) nurses and patients families in Peking Union Medical College Hospital. Methods The Department of Critical Care Medicine of Peking Union Medical College Hospital distributed questionnaires to 143 nurses working in the department, and scored whether nurses had negative experience in communicating with patients'families, confidence in building harmonious relationship and the need for training. Results A total of 138 questionnaires were obtained. 46.4% of nurses in ICU think they sometimes or often encounter negative experiences in their work. Only 30.4% of nurses were very confident or confident in their communication skills. As much as 83.3% of nurses believed that they should be trained in communication with patients family members. Conclusions Communication training is very important for intensive care nurses.

Comparison of Chinese and foreign medical humanities teaching methods

GUO Xu-fang, LIU Jun-xiang, LIU Hui, HE Zhong

2020, 40(1):  132-135. 

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With the advance of medical education reform, the teaching methods of medical humanities courses in the international arena are constantly innovating and remodeling. Learning the new teaching methods from foreign medical humanities has a positive impact on promoting domestic education of medical humanity. This article tries to showed what can be learn from medical education by comparing the teaching methods of medical humanities courses in several American universities and British universities with several colleges in China.

Application of case based learning rounds with PACS in the training of radiology clinicians

PEI Xin-long, LANG Ning, ZHANG Qi, YUAN Hui-shu

2020, 40(1):  136-140. 

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Objective To explore the value of case-based learning rounds(CBL) with application of PACS in the training of diagnostic capacity of radiology clinicians. Methods Peking University third hospital radiology department carried out the CBL rounds with application of PACS for 41 CT training clinicians in 2018, then compared with 39 CT training clinicians (without CBL rounds) in 2017. Ten reports written by each clinician(reported during the last training monthes) were randomly inspected from the PACS report system before and after the CBL rounds.These reports were scored, the excellent and good rate was calculated. The scoring criteria include accurate description of lesion imaging features, standardized report content, reasonable diagnosis logic, answering questions from clinical application form, making reasonable suggestions for further diagnosis and treatment, and no missed diagnosis. At the same time, questionnaires were used to evaluate the subjective feelings of training clinicians for CBL rounds, including comprehensive understanding of diseases, improvement of clinical analysis ability, reading images ability, and learning interest, and help of PACS for learning, expressed by percentage. Results 1)After CBL teaching, the report quality scores of clinicians increased(P＜0.01). 2)Satisfaction rate of clinicians was 90.2%,68.3%,80.5%, 75.6% and 80.5% respectively. Conclusions CBL rounds with application of PACS system is an effective method to train the diagnosis capacity of radiology clinicians.

Investigation and analysis of multi-grades psychological status of eight-year undergraduates in clinical medicine

LI Ning-ning, LI Hang, SHI Xiao-chun, JIANG Yi-nan, ZHAO Jun

2020, 40(1):  141-144. 

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Objective To investigate the mental health status of eight-year medical undergraduates in different academic years and to analyze the influencing factors, so as to put forward countermeasures to improve the mental health of students. Methods Questionnaires were used to investigate the mental health status of undergraduate medical students in grades 5 to 7 in our university. The differences among groups were analyzed according to different grades. The statistical methods of Chi-square test and Fisher exact test were used for data analysis. Results The recovery rate of the questionnaire was 96.5%, and 164 qualified questionnaires were obtained. Fourteen students (8.5%) had depression and 28(17%) had anxiety problems. Fifty-seven cases (34.76%) needed the help of self-rating questionnaire and 110 cases (67.07%) needed the help of depression and anxiety counseling. The proportion of demand for depression/anxiety counseling among sixth grade medical students was significantly higher than that of fifth grade (82% vs 50%; P＜0.05) and seventh grade (82% vs 57%; P＜0.05). As much as 86.67% of the seventh grade students had self-rating psychological questionnaires, higher than the fifth grade group (64.29%, P＜ 0.05) with no significant difference with the sixth grade group. Conclusions The mental health status of eight-year medical students in different learning stages is different, and there are emotional adjustment disorders different degrees. Effective measures they need to support and psychologically intervention.