Table of Content

05 June 2022, Volume 42 Issue 6

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Original Articles

Efficient editing of mouse miR let-7a based on CRISPR/Cas9 system

ZHOU Lan, ZHANG Sheng-gui, HU Zu-liang, WANG Tian-xian, YUE Peng-peng, YU Hong-hao

2022, 42(6):  857-863.  doi:10.16352/j.issn.1001-6325.2022.06.023

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Objective To establish effective CRISPR/Cas9 system targeting mouse miR let-7a. Methods Two pairs of dual-sgRNAs sequences were designed according to the coding sequences of two coding sites let-7a-1 and let-7a-2 of mouse let-7a respectively. The targeting potency of four pairs of sgRNA was analyzed through the construction of sgRNA expression vector, cotransfection of N2a cells, drug screening. Positive cells' PCR products sequencing and TA clone sequencing. Results The four pairs of targeting vectors played a role in the cells, and there were base insertions or deletions at the targeting sites, and the Sanger sequencing peaks of RCR products at the drug screening positive cells showed nested peaks at all four targeting sites.The results of TA cloning sequencing showed that the targeting efficiency of the four pairs of dual-sgRNAs targeting sequences designed in this study was 42.10%, 62.50%, 52.63% and 47.37% respectively. Conclusions The results showed that the four pairs of dual-sgRNAs-directed sequences designed in this study deavaged miR let-7a by inducing Cas9 protein effectively and thus generated a mutating effect in mice. Our study laid a foundation for further exploring of let-7a targets and elucidating the mechanism of its' anti-cancer function.

Epigenetic regulation mechanism of EZH2 gene in hepatocellular carcinoma

ZHAO Yu-shan, JI Meng-die, DONG Yu-cheng, GUO Xin, SONG Bo-yuan, CHEN Yang

2022, 42(6):  864-870.  doi:10.16352/j.issn.1001-6325.2022.06.026

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Objective This study combined public data analysis with biological experiment using the tissues of hepato-cellular carcinoma to explore the epigenetic regulation mechanism of EZH2 gene which is potentially related to the progression of hepato-cellular carcinoma. Methods Using the TCGA's public clinical data of hepato-cellular carcinoma, the relation of EZH2 gene expression with tumor progression as well as prognosis was analyzed. Subsequently, the effect of EZH2 inhibitor GSK343(10 μmol/L) on the proliferation of hepato-cellular carcinoma cell line HepG2 was further analyzed by ATP cell activity assay and crystal violet assay. Then, the differentially expressed genes between EZH2 high expression group and low expression group in TCGA samples were analyzed. After that, the ChIP-seq public data of ENCODE was used to analyze the histone modification at the transcription starting site of EZH2 downstream target gene. Finally, RT-qPCR was used to detect the gene expression of downstream target genes regulated by EZH2. Results EZH2 gene-expression was significantly correlated with the progression of hepato-cellular carcinoma from stage 1 to stage 3(P＜0.05). Survival analysis showed that the high expression of EZH2 gene was significantly correlated with the poor prognosis of hepatocellular carcinoma(P＜0.05). The EZH2 inhibitor could significantly inhibit the proliferation of hepatocellular carcinoma cell line HepG2 in vitro (P＜0.05). Histone modification H3K27me3 was enriched at the transcription start site of EZH2 binding gene. There was 163 genes significantly different in EZH2 high expression group and low expression group and had EZH2 binding sites at their transcription start sites. The experiment verified that ADRA1A was a downstream target gene of EZH2. Conclusions EZH2 gene may affect expression of its downstream target genes through epigenetic regulation in hepato-cellular carcinoma.

Peripheral blood miR-200c level predicts the occurence of acute graft-versus-host disease after allogeneic hematopoietic stem cell transplantation

WU Kun, NIE Bo, LI Yun-tao, YANG Jin-rong, CHENG Shen-ju, LI Yan-hong, SHI Ming-xia

2022, 42(6):  871-875.  doi:10.16352/j.issn.1001-6325.2022.06.001

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Objective To investigate the predictive function of miR-200c level in peripheral blood on acute graft-versus-host disease (aGVHD) after allogeneic hematopoietic stem cell transplantation. Methods Fifty-one patients who received allo-HSCT in hematology department of our hospital from January 2014 to October 2018 were selected as the research subjects. According to the principle of whether agVHD occurred after transplantation, 33 patients were in the experimental group (with aGVHD) and 18 patients were in the control group (without aGVHD). RT-qPCR was used to detect miR-200c in two groups and the differences in miR-200c expression between the two groups were analyzed. Results The transplantation was successful in both groups, and the hematopoietic function was restored. The recovery time was 18 days, among which 33 patients developed aGVHD and 18 patients did not develop aGVHD, with an incidence of 64.71%. The expression of miR-200c in experimental group was lower than that in control group before transplantation and 7, 14, and 72 d after transplantation (P＜0.05). MiR-200c in peripheral blood after the occurrence of aGVHD was significantly lower than that before the occurrence(P＜0.001). Compared with aGVHD Ⅱ, the expression of miR-200c in peripheral blood of aGVHD Ⅱ patients was down-regulated (P＜0.05); as compared with aGVHD Ⅲ patients, the expression of miR-200c in peripheral blood of aGVHD Ⅲ patients was down-regulated (P＜0.001); the 2-year survival rate in the high expression group of miR-200c was higher than that in the low expression group (P＜0.05). Conclusions The reduced level of miR-200c is associated with the occurrence of aGVHD after allogeneic hematopoietic stem cell transplantation. With the increase of the severity of aGVHD the expression level of miR-200c is decreased, and the survival rate of patients with low miR-200c expression is low, so the expression of miR-200 is a potential marker for early diagnosis and prediction of aGVHD.

Analysis of the immune-related molecule expression and immune infiltration in mouse breast cancer stem cells

LI Yu-yang, LI Zhao-xuan, LUO Yun-ping, CHEN Chong

2022, 42(6):  876-882.  doi:10.16352/j.issn.1001-6325.2022.06.002

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Objective To investigate the expression of immune-related molecules and the infiltration of immune cells from mouse breast cancer stem cells (BCSCs). Methods Mouse BCSCs of 4T1 and 4T07 were enriched by spheroid culturing; the expression of immune checkpoint-and antigen presentation-associated genes in BCSCs was detected by RT-qPCR; the proportion of various types of immune cells in spleen or tumor tissue of orthotopic tumor model mouse was detected by flow cytometry. Results The mouse BCSCs 4T1 and 4T07 were successfully enriched by spheroid culturing; the expression of immune checkpoints-and antigen presentation-associated genes were significantly different as compared with the adherent cancer cells; the proportion of both CD4⁺ and CD8⁺ T cell increased. However, the proportion of macrophage (CD11b⁺/F480⁺) was decreased in tumors formed by spheroid cells as compared with that in the tumors formed by adherent cells. Conclusions The expression of immune checkpoint-and antigen presentation-associated genes in BCSCs were significantly different and the infiltration of T cells increased in tumors formed by BCSCs.

Prokaryotic expression of glucans binding protein and application for detecting invasive fungal infection

ZHU Li-ru, HUANG You-ming, ZHANG Hong-xiang, ZHOU Fa-you, TANG Xiao-lei

2022, 42(6):  883-889.  doi:10.16352/j.issn.1001-6325.2022.06.007

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Objective To explore the potential application of glucans binding protein (GBP) in the detection of soluble glucans from serum sampled from patients with invasive fungal infection (IFI). Methods The prokaryotic plasmid pET30a-GBP252-668 coding for 252 to 668 amino acid in GBP as an active domain of GBP of Limulus polyphemus was constructed. The recombinant GBP252-668 was expressed and purified.Then a double GBP252-668 sandwich method was developed to detect (1,3)-β-D-glucans in the serum of patients with IFI. Meanwhile, the results were compared with G-test. Results Recombinant GBP252-668 (molecular weight is about 46 ku) was obtained and the double GBP252-668 sandwich method was established and optimized via chessboard method, the concentrations of GBP252-668 were 8 and 16 g/L for coating and detection respectively. The serum sampled from 48 patients with IFI and from 48 healthy donors were examined.and results were analysed. The sensitivity and specificity of double GBP252-668 sandwich method were 91.67% and 89.58% respectively and the sensitivity was non-inferior to G-test, and specificity of double GBP252-668 sandwich method was even higher than that of G-test (79.17%). Conclusions The GBP252-668 sandwich method is a potential detection method for IFI diagnosis. The method is non-inferior to G-test and its specificity is even higher than G-test. It is expected to be a new commercially available kit for IFI detection.

Liver pathological changes and hepatocyte acquisition method of model mice with non-alcoholic fatty liver disease

SHI Dong-xue, YUAN Jia-qi, DING Bao-feng, WAGN Xiao-shuang, YU Jia

2022, 42(6):  890-895.  doi:10.16352/j.issn.1001-6325.2022.06.029

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Objective To detect the pathological changes in the liver of mice with non-alcoholic fatty liver disease (NAFLD) and explore an experimental method to obtain a large number of primary hepatocytes with high activity and purity from the mice with NAFLD. Methods A total of 60 mice were randomly divided into control group (chow diet, CD) and high-fat group (high-fat diet, HFD) and had been fed separately for 20 weeks. The body weight of the mice was regularly measured and the pathological characteristics of the liver tissue were observed by hematoxylin-eosin (HE) staining and Sirius red staining. After 15 weeks of high-fat diet, 6-8 mL collagenase Ⅳ were added for digestion to obtain mixed hepatocytes, which were spread on 40% Percoll separation solution and subjected to density gradient centrifugation to obtain purified hepatocytes. The viability of the cells was determined by trypan blue staining, and the morphology was observed by microscope. Results With the prolongation of modeling time, the body weight of mice in the HFD group were gradually increased, which was significantly higher than that in the CD group. In the HFD group, the liver of mice gradually developed fatty degeneration, and liver fibrosis could be detected at week 20. At week 15, about 5×10⁷-7×10⁷ hepatocytes could be obtained from each NAFLD mouse with cell viability ≥85% and cell purity ≥95%. Cells were round or elliptical, with smooth edges and few clusters. The number and quality of cells obtained were significantly improved by optimizing conditions such as accurate temperature control, reduced the concentration of collagenase, and the use of low-speed centrifugation and low-speed density gradient centrifugation. Conclusions The NAFLD mouse model was successfully established and the pathological characteristics of liver in different disease stages were revealed. An efficient and stable method for obtaining primary hepatocytes of NAFLD mice was established. The number and activity of hepatocytes obtained met the requirements of most experiments.

Establishment　and identification of an ovarian cancer cell strain stably expressing CDC50A

HAN Tian-tian, YIN Jie, ZENG Jing, JIN Ying, LI Yan, PAN Ling-ya

2022, 42(6):  896-901.  doi:10.16352/j.issn.1001-6325.2022.06.003

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Objective To establish a stable-expressing human cell cycle regulatory protein 50A (CDC50A) SKOV3 cell strain expressing vector containing CDC50A by molecular cloning. Methods CDC50A was introduced into the expression vector pLVX-IRES-GFP by molecular cloning method. The pLVX-CDC50A-GFP expression plasmid was packaged by lentivirus and stably transfected into SKOV3. SKOV3 cell strain with high-expressing CDC50A was established. The expression of Flag protein was analyzed by flow cytometry, Western blot and immunofluorescence. Results The construction of pLVX-CDC50A-GFP expressing vector was confirmed by gel electrophoresis and endonuclease digestion. The efficiency of infection was observed qualitatively under fluorescence microscope and detected quantitatively by flow cytometry as 86.5%. Flag-tag protein was detected by Western blot to present only in the positive group. Conclusions The SKOV3 cell strain was established by infecting with lentivirus of pLVX-CDC50A-GFP, laying a foundation for the mechanism study of CDC50A in ovarian cancer.

Optimize nearly PAM-less adenine base editor to improve editing efficiency

ZHOU Xiao-yu, WANG Xiao-yue

2022, 42(6):  902-907.  doi:10.16352/j.issn.1001-6325.2022.06.028

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Objective To explore the effects of replacing different components of PAM-less adenine base editor (ABE) on its editing efficiency. Methods ABEmax was replaced with Tad-8e in SpRY-ABEmax to construct SpRY-ABE8e. Then the connecting peptide 3xGGS-XTEN-3xGGS between Cas9 protein variant SpRY and Tad-8e was shortened to PAPAPA. ABEs and sgRNA expression plasmids were co-transfected into HEK 293T cells. After 24 hours, cells were incubated with 1.5 μg/mL puromycin. After 72 hours, the cells expressing green fluorescent protein were flow sorted and the cell genome was extracted and the expected editing sites were amplified by PCR technology. Sanger sequencing was performed and Beat tool was used to estimate the editing efficiency of each site. Results Compared with control group, SpRY-ABEmax, SpRY-ABE8e has higher editing efficiency and expanded editing window(P＜0.05); editing window of SpRY-ABE8e can be limited to a certain extent while maintaining editing efficiency by using the shortened linker peptide. Conclusions The editing efficiency can be improved by replacing different components of PAM-less ABE, which will provide more candidate tools for gene therapy in the future.

Gypenoside alleviates injury of rat adrenal pheochromocytoma cell line PC12 induced by lipopolysaccharide

JIANG Xiu-fang, GENG Ya-nan, YANG Sheng-hong, WEI Ping, ZHAO Ming, ZHU Ling-ling

2022, 42(6):  908-912.  doi:10.16352/j.issn.1001-6325.2022.06.024

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Objective LPS-treated PC12 cells were used to establish neuron inflammation model. RT-PCR was used to detect the mRNA of cytokines. CCK-8 assay was used to check determine cell viability. Western blot was used to detect NF-κB pathway. Methods LPS-treated PC12 cells were used to establish neuron inflammation model. RT-PCR was used to detect the mRNA of cytokines. CCK-8 assay was used to check determine cell viability. Western blot was used to detect NF-κB pathway. Results After 12 hrs treatment of PC12 cells with different doses of LPS (0, 100, 300 and 1 000 ng/mL), the mRNA level of TNF-α,IL-6 and IL-1β all increased. The cells pre-treated with different doses of gypenoside (30, 100 μg/mL) and then treated with LPS for 24 h, cell viability decreased to 85.7%. However, 30 and 100 μg/mL gypenoside pre-treatment resulted in cell viability increased to 96.2% and 97.9% respectively. Gypenoside also inhibited LPS-induced cell inflammation as indicated by increased mRNA level of TNF-α, IL-6 and IL-1β caused by LPS stimulation. LPS increased the phosphorylation of p65 (p-p65) and the phosporylation was also inhibited by gypenoside treatment. Conclusions Gypenoside alleviates LPS-induced inflammation and then protects PC12 cells from injury induced by LPS through inhibiting NF-κB pathway.

Effect of lncRNA SNHG14 on oligomeric Aβ-induced injury in rat neural cell line PC12 through regulating miR-142-5p

SHEN Jie, YAN Jie, ZHONG Ming, PANG Rui-juan, LUO Yong-jie

2022, 42(6):  913-919.  doi:10.16352/j.issn.1001-6325.2022.06.022

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Objective To investigate the effect of lncRNA SNHG14 targeting at miR-142-5p on the neuronal damage induced by oligomeric Aβ. Methods PC12 cells were divided into control group, model group, model+si-NC group, model+si-SNHG14 group, model+miR-NC group, model+miR-142-5p group, model+si-SNHG14+anti-miR-NC group, and model+ si-SNHG14+anti-miR-142-5p group. The level of SNHG14 and miR-142-5p was calculated by RT-qPCR. CCK-8 method and flow cytometry were applied to detect inhibition rate on cell proliferation and apoptosis rate. ELISA method was used to detect TNF-αand IL-6 in the culture. Dual luciferase report assay was used to find the targeting relationship between SNHG14 and miR-142-5p. Results SNHG14 was highly expressed (P＜0.05)while miR-142-5p was lowly expressed (P＜0.05) in plasma of AD patients. Compared with the control group, the SNHG14 level, cell proliferation inhibition rate, and apoptosis rate in the model group were significantly increased (P＜0.05), the level of miR-142-5p was significantly reduced (P＜0.05), and the levels of TNF-α and IL-6 in the culture medium were significantly increased (P＜0.05), IL-10 level was significantly decreased (P＜0.05). Compared with the model+si-NC group, the cell proliferation inhibition rate and apoptosis rate in the model+si-SNHG14 group were significantly reduced (P＜0.05), the level of TNF-α and IL-6 in the culture medium was significantly reduced (P＜0.05), IL-10 level was significantly increased(P＜0.05). Compared with the model+miR-NC group, the cell proliferation inhibition rate and apoptosis rate in the model+miR-142-5p group were significantly reduced (P＜0.05), and the levels of TNF-α and IL-6 in the culture medium were significantly reduced (P＜0.05) and IL-10 level was significantly increased (P＜0.05). miR-142-5p was a direct target of SNHG14. Compared with the model+si-SNHG14+ anti-miR-NC group, the cell proliferation inhibition rate and apoptosis rate in the model+ si-SNHG14+anti-miR-142-5p group were significantly increased(P＜0.05), and the levels of TNF-α and IL-6 in the culture medium were significantly increased (P＜0.05), IL-10 level was significantly decreased (P＜0.05). Conclusions Silencing SNHG14 may alleviate oligomeric Aβ-induced neuronal damage by targeting at and promoting miR-142-5p expression.

Nuclear transcription factors NFYB and NFYC promote terminal erythroid differentiation

YU Dong-lin, YANG Xi, YIN Jia-ying, LIU Xue-hui, LYU Xiang

2022, 42(6):  920-926.  doi:10.16352/j.issn.1001-6325.2022.06.006

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Objective To predict potential regulators of terminal erythroid differentiation by single-cell transcriptome data mining, and then focus on exploring the expression pattern and function of nuclear transcription factors NFYB and NFYC in terminal erythroid differentiation. Methods Using published single-cell transcriptome data of human terminal erythropoiesis, SCENIC analysis was performed to predict stage-specific transcription regulators. The expression levels of two predicted regulators, NFYB and NFYC, in different stages of human and mouse terminal erythropoiesis were first analyzed in public bulk RNA-seq data; cells among different stages of terminal erythroid differentiation in mouse fetal liver were sorted by flow cytometry, NFYB and NFYC expression levels were then detected by RT-qPCR. In addition, the expression of NFYB and NFYC was inhibited by lentivirus-mediated shRNA interference and the efficiency of terminal erythroid differentiation and enucleation was examined. Results NFYB and NFYC were potential regulators at early stage of terminal erythropoiesis by SCENIC analysis. Their expression level was high at early stage of both human and mouse terminal erythropoiesis and then decreased at the last stage. Knockdown of Nfyb and Nfyc in mouse fetal liver derived erythroid cells significantly inhibited terminal erythroid differentiation and enucleation (P＜0.05). Conclusions NFYB and NFYC promote the terminal erythroid differentiation.

Okadaic acid induces damage of human neuroblastoma cell line SH-SY5Y

LU Ya-lan, ZHOU Li, HAN Yun-lin, WANG Ke-wei, QIN Chuan

2022, 42(6):  927-932.  doi:10.16352/j.issn.1001-6325.2022.06.008

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Objective To investigate the effect of okadaic acid (OA)-induced phosphorylation of neuron micro-tubule associated protein tau (Tau) on the damage of human neuroblastoma SH-SY5Y cells. Methods SH-SY5Y cells were incubated with different concentrations (20 and 40 nmol/L) of OA for 0, 12, 24, 48 and 72 hours followed by microscopy for cell morphology.Cell ultrastructure was observed by transmission electron microscope and cell apoptosis was detected by TUNEL assay. The protein expression of p-Tau and caspase3 was analyzed by Western blot and mRNA level of inhibitor of apoptosis family proteins (IAPs) was determined by RT-qPCR. Results OA induced phosphorylation of Tau protein in SH-SY5Y cells as compared to the result from control goup (P＜0.001), promoted cell damage and apoptosis (P＜0.001).OA also injured mitochondrial structure in a concentration and time dependent manner. It can also up-regulate IAPs expression significantly (P＜0.05). Conclusions The aberrant phosphorylation of Tau protein in SH-SY5Y cells induced by OA may lead to cell injury and apoptosis. Inhibition of endogenous apoptotic pathways or up-regulation of IAPs family proteins provides potential targets for AD prevention and treatment.

High-throughput screening of inhibitors of NMNAT1 and their tumor-targeting functions

SHA Wu-ga, LI Jun

2022, 42(6):  933-939.  doi:10.16352/j.issn.1001-6325.2022.06.025

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Objective To find new drug candidates for cancer treatment by screening inhibitors of NMNAT1 using recombinant human NMNAT1 expressed in E. coli system. Methods The human NMNAT1 was inserted into the pET21b(+) vector and the recombinant plasmid was transformed into E. coli BL21(DE3) strain for over expression of NMNAT1 protein. The enzymatic activity of NMNAT1 was measured by a three-step coupled fluorescence assay, and 2 280 chemicals from a nature compounds library were screened using this assay for potential inhibitors of NMNAT1. The potential target compounds that inhibited NMNAT1 were verified at the cellular level and tested for tumor-killing effect. Results The recombinant human NMNAT1 proteins with high purity and activity were obtained and 6 kinds of high-efficiency NMNAT1 inhibitors were identified. One of the them, fraxetin, efficiently reduced the NAD⁺ content of breast cancer cells and promoted cell apoptosis. RNA-seq analysis of NMNAT1 knock-down cells revealed the function to enhance apoptosis which is consistent with the cellular effect of NMNAT1 inhibitor. Conclusions Fraxetin is a high-efficiency NMNAT1 inhibitor to promote apoptosis of breast cancer cells.

Down-regulation of lncRNA RHPN1-AS1 inhibits proliferation and migration of human bladder cancer cell line T24

BAI Bing, ZHANG Hai-fang, YANG Qing-liang, QIN Yue, ZHOU Chen-long, SONG Ge, WANG Ying

2022, 42(6):  940-944.  doi:10.16352/j.issn.1001-6325.2022.06.004

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Objective To explore the effect of interfering with lncRNA RHPN1-AS1 on the biological behavior of bladder cancer cell T24 and its mechanism. Methods RT-qPCR method was used to detect the expression of RHPN1-AS1 and miR-149-3p by bladder cancer tissues and adjacent tissues.si-NC,si-RHPN1-AS1, si-RHPN1-AS1,anti-miR-NC, si-RHPN1-AS1 and miR-149-3p inhibitor were transfected into bladder cancer cell T24. The expression of RHPN1-AS1 and miR-149-3p in T24 cells were examined by RT-qPCR.MTT, plate colony formation experiment, scratch experiment, flow cytometry were used to detect cell proliferation, migration ability and cell apoptosis rate respectively. The dual luciferase reporter experiment was used to detect the targeting relationship between RHPN1-AS1 and miR-149-3p. Bax and Bcl-2 protein expression was assessed using Western blot. Results Compared with adjacent tissues, the expression of RHPN1-AS1 in bladder cancer tissue was increased(P＜0.05) and the expression of miR-149-3p was decreased (P＜0.05). Compared with the si-NC group, transfection of si-RHPN1-AS1 significantly reduced cell proliferation, scratch healing rate and the protein level of Bcl-2(P＜0.05), reduced the number of clones (P＜0.05)and increased apoptosis rate and the protein level of Bax(P＜0.05). Compared with the si-RHPN1-AS1+anti-miR-NC group, transfection of si-RHPN1-AS1 and miR-149-3p inhibitor significantly increased cell proliferation, scratch healing rate and the protein level of Bcl-2 (P＜0.05) increased the number of clones formed (P＜0.05), reduced the apoptosis rate and the protein level of Bax (P＜0.05). Conclusions Interfering with the expression of RHPN1-AS1 may inhibit the proliferation and migration of bladder cancer cells and may promote cell apoptosis by up-regulating miR-149-3p.

Clinical Sciences

Depressive symptoms increase the risk of falls in elderly Chinese

LIU Yan-li, ZHAO Rui-xue, PAN Yan, MA Ya-jun, ZHAO Tun, JIANG Xiao-yan, LI Shu-juan

2022, 42(6):  945-949.  doi:10.16352/j.issn.1001-6325.2022.06.027

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Objective To investigate whether depression is a potential risk factor for falls in the elderly in China. Methods Data were collected from the Subcohort of Rugao Longevity and Aging Study. Depression in the elderly was assessed by the GDS-15(Geriatric Depression Scale), and the occurrence of falls in the subjects at baseline in 2014 and the second follow-up in 2017 were collected and recorded. According to the depression status, subjects were divided into depression group and normal group, and the general information and occurrence of falls were compared between the two groups. Logistic regression was used to analyze whether depression increased the risk of falls in older adults. Results In the cross-sectional study of 1 548 subjects, the incidence of falls was 8.3%. The proportion of subjects in the “fall” group was higher in older age, lower education level and cognitive impairment. Compared with the normal group, the depressed group had a higher proportion of women, farmers, less education, unmarried, non-smokers, non-drinkers and cognitively disabled people. After adjusting for confounders, depression was associated with an increased risk of falls at baseline and 3 years later in older adults (P＜ 0.05). Conclusions Depression is a potential risk factor for falls among the elderly in the Chinese community.

Application of low-dose esketamine combined with dexmedetomidine in anesthesia for elderly patients with percutaneous vertebroplasty

REN Hai-qiang, LI Lei, YANG Wang-yan, DU Juan, JIANG Dan-dan, YAN Li, FU Run-qiao

2022, 42(6):  950-954.  doi:10.16352/j.issn.1001-6325.2022.06.020

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Objective To investigate the effects of low-esmketamine combined with dexmedetomidine on intra-venous analgesia, sedation in elderly patients undergoing percutaneous vertebroplasty (PVP). Methods A total of seventy-two elderly patients aged 70-90 years old who underwent PVP in Beijing Chuiyangliu hospital between March to November 2021 were enrolled in this study. Patients were assigned randomly by a random number table method into controll group (SF-DEX, n=36) and test group (ES-DEX, n=36) for the seddative analgesia anesthesia. All patients were accepted a 0.5 μg/kg dexmedetomidine by pump into venous within 10 min, the other 5 μg sufentanil was gived venously and then 0.5 μg/(kg·h) dexmedetomidine by pump in SF-DEX. 0.1 mg/kg esketamine was gived venously and then 0.1 mg/(kg·h) by pump in EK-DEX. The changes of circulatory indexes systolis arterial pressure(SAP),diastolis arterial pressure(DAP),mean arterial pressure(MAP),heart rate(HR), saturated oxygen (SpO₂), BIS indexes, pain intensity (VAS), Ramsay score for sedation level, blood pressure variation, arrhythmia and decrease range of SpO₂, were measured respectively at four time points before administration (T1), at time of operation start (T2), filling bone cement (T3), at the end of operation (T4) between two groups. Results SAP,MAP was significantly lower at T3 than that at T1 (P＜0.05) in SF-DEX; the Ramsay score was increased at T2 and T3 (P＜0.05).The VAS score in EK-DEX group was significantly lower than that in SF-DEX group at T2 and T3 (P＜0.05); the Ramsay score in EK-DEX group was significantly increased than that in SF-DEX group at T3 (P＜0.05). The incidences and hemodynamic adverse effects such as hypotension, bradycardia and desaturation, were not significantly different between the groups. Conclusions Low-dose esketamine combined with low-dose dexmedetomidine for anesthesia produces satisfactory analgesic and sedation effect in elderly patients with PVP without obvious complications. It is a safe and feasible method.

Serum screening for Down's syndrome in the second trimester of pregnancy is associated with adverse pregnancy outcomes

WANG Zhan-yong, XU Jing, LIANG Yan, YUE Yu-hong

2022, 42(6):  955-959.  doi:10.16352/j.issn.1001-6325.2022.06.019

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Objective To explore the association between screening for Down's syndrome and adverse pregnancy outcomes. Methods Retrospective cohort study was conducted in Beijing Chaoyang Hospital covering pregnant 7 682 pregnant women from Mar 31, 2014 to Apr 29, 2021 in a continuum period. Results The prevalence of high screening risk was 6.40% (492 cases) and the incidence of adverse pregnancy outcome was 1.30%(100 cases). The incidence of adverse outcome (4.27%) in high risk group was significantly higher than that in low risk group (1.10%). After adjusting for diabetes, family history of hereditary diseases, smoking and drinking alcohol, the incidence odds of adverse outcomes for high screening risk subjects was 4.022 times (95% CI:2.405-6.442) as that for low risk. The high Down's syndrome risk score meant high incidence of adverse outcomes(P＜0.001). The incidences of malformations, chromosome abnormalities and stillbirth for high screening risk subjects were higher than those for low risk. The incidence odds of adverse outcomes increased to 1.019 times with the increase of 1 ng/mL for AFP and 2.258 times with the increase of 1 for AFP MOM. The incidence odds of adverse outcomes declined to half with the increase of 1 ng/mL for uE3 or uE3 MOM. Conclusions Down's syndrome screening high risk is indicators for organ malformations, limb deformity, chromosome abnormalities and still birth. High level of AFP and low level of uE3 may increase the occurrence of adverse pregnancy outcomes.

Mini Reviews

Research progress of dioscin in prevention and treatment of primary osteoporosis

XIE Xing-wen, ZHONG Jian-chun, LI Ding-peng, HUANG Rui, LI Ning, GUO Yun-long, ZHAI Bo-yu

2022, 42(6):  960-963.  doi:10.16352/j.issn.1001-6325.2022.06.021

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Dioscin has strong anti infection, anti-virus, lipid-lowering, anti-cancer, hypoglycemic, antithrombotic, anti-aging and liver protective activities. It can protect cardiac cells from ischemia/reperfusion injury by delaying oxidative stress, and become one of the complementary and alternative clinical drugs for the treatment of coronary heart disease and angina pectoris. In recent years, studies have shown that dioscin can prevent osteoporosis(OP) by inhibiting RANKL induced osteoclast differentiation and promoting osteoblast differentiation. This paper reviews the research progress of dioscin in the prevention and treatment of osteoporosis in the aspects of inhibiting osteoclasts, promoting osteogenesis and related signaling pathways, in order to provide ideas for the development of a nutrition-medicine hybyrid strategy in the prevention and treatment of OP.

Research progress of inflammatory cytokines in the diagnosis and treatment of amyotrophic lateral sclerosis

LI Xiao-guang, YANG Lu, LIU Xu-dong, JIA Xin-miao, CHENG Yan-fei, CUI Li-ying

2022, 42(6):  964-968.  doi:10.16352/j.issn.1001-6325.2022.06.012

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Amyotrophic lateral sclerosis (ALS) is a multifactor, multiple genes disease which involving a variety of mechanisms. Neuroinflammatory response is one of the mechanisms. Many inflammatory cytokines are involved. It may be used in clinic as biomarkers and therapeutic targets. Many proteins encoded by mutant genes can damage the function of immune system, lead to immune dysfunction and drive the inflammatory response of central nervous system and peripheral immune system. There are many kinds of inflammatory cytokines with great diversification in their levels which influenced by the stage, type and comorbidity of the disease, the results are not consistent at present, and there is no single inflammatory cytokine identified as a specific marker. Inflammatory cytokines have a certain prospect as therapeutic monitoring markers and targets.

Advantages, disadvantages and research progress on strategies for screening and identifying functional genes

LIU Xu, CHANG De, WANG Jun-feng

2022, 42(6):  969-974.  doi:10.16352/j.issn.1001-6325.2022.06.010

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The screening and identification of functional genes is a key step to deeply understand various biological phenotypes. Forward genetics including traditional genetics, genome-wide association analysis, the comparison of gene expression profiles and bioinformatics, and reverse genetics including cDNA library, RNA interference, CRISPR-Cas9 technology, insertion mutation as well as model organisms have been applied to screening functional genes. Different methods have their own advantages and disadvantages and technology combination to screen and to identify functional genes is a good choice of strategy.

Literature publication by clinical laboratory in Tibet Autonomous Region

BIAN Zhen, Nimadunzhu, QIN Xu-zhen

2022, 42(6):  975-978.  doi:10.16352/j.issn.1001-6325.2022.06.011

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Objective To investigate the publications from clinical laboratory in Tibet Autonomous Region in recent 40 years and to summarize the scientific research status. Methods With the institution name of “Tibet” and “clinical laboratory”, we retrieved 297 publications from Wanfang database from 1980 to 2020,using EXCEL software to analyze the results from annual publications, source journals, distribution of institution, scientific research funds, proportions of first author from Tibetan, keyword frequency, and etc. Results Total of 297 publications were included from 1980 to 2020 in this study. The number of articles has gradually increased since 2001. Core journals accounted for less than 30%. The publications were mainly from the two major tertiary hospitals in Tibet; 1% of the articles have got supported by scientific research funds. The proportion of papers with first author from Tibet only takes 28%. Conclusions In recent decades, the development of scientific research by clinical laboratory in Tibet has improved significantly, but there is still space to fill in future with effort.

Research progress on the correlation between iodine and metabolic syndrome

XU Xiao-shuang, MIN Hang, YE Yue, GUO Meng-ran, ZHEN Dong-hu

2022, 42(6):  979-982.  doi:10.16352/j.issn.1001-6325.2022.06.009

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Metabolic syndromes (Mets) is a high-risk factor for a series of cardiovascular diseases. Changes in iodine nutritional status can directly mediate the occurrence of insulin resistance, affect glucose and lipid metabolism and the regulation of blood pressure and also,may contribute to the pathogenesis of Mets by regulating thyroid function. Exploring the role of iodine in the occurrence and development of Mets can provide new ideas for the prevention and treatment of Mets.

Up-date situation of research on the effect of diet on gut microbiota-immune axis and potential relationship to hypertension

WANG Zhen-hua, LI Chao-sheng

2022, 42(6):  983-987.  doi:10.16352/j.issn.1001-6325.2022.06.005

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Gut microbiota and gut-derived metabolites are known to respond to fluctuations in dietary composition. Diet-induced shifts in microbiota composition have profound effects on immune regulation, which can also contribute to hypertension. Recent years, studies have explored changes in gut microbiota and immune inflammatory in response to diet, as well as their association with hypertension in order to provide new targets and new ideas for the prevention treatment of hypertension.

Medical Education

Surgical education based on combination of robotic-arm assisted total knee arthroplasty design and 3D-printing simulation

HUANG Ying, LUO Wen-hao, FAN Yu, LIN Jin

2022, 42(6):  988-993.  doi:10.16352/j.issn.1001-6325.2022.06.014

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As compulsory content for orthopedic graduate students, total knee arthroplasty (TKA) is characterized by requirements of surgical accuracy and so is difficult to train since there is limited opportunity for young surgical trainees especially design of surgical operation and long term training to get experience. Traditional teaching of TKA for graduates is embarrassed by these barriers. Recent years, new techniques such as robotic-arm assisted surgery and 3D-printing technology have been extensively used in clinical training. For example the hybrid model with combined surgical design in virtue of robotic system and vivid simulation produced by 3D-printing for training. This paper reviewed is current situation in this field and evaluated the feasibility in clinical application

Evaluation of the Q-TRUST clinical communication training course in training new staff of emergency department

LIU An-lei, ZHU Hua-dong, PENG Hua, GUO Na, HAN Hong, YANG Jing, LI Yi

2022, 42(6):  994-998.  doi:10.16352/j.issn.1001-6325.2022.06.016

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Objective To evaluate the application effect of Q-TRUST clinical communication training course in the training recruits of emergency department. Methods The experimental group was selected from new staff of the emergency department at Peking Union Medical College Hospital from October 2019 to September 2021.Totally 120 recruits from October 2017 to September 2019 were selected as the control group with 60 in each. The control group was adopted traditional centralized training, and the experimental group was adopted Q-TRUST clinical communication training. The basic information of the students in the two groups was collected, and the communication theory assessment was conducted after the training. The results of patient satisfaction and dispute complaints in the depart-ment within 2 years after the training were collected. The training effect evaluation of experimental group was obtained through questionnaire survey. Results There was no significant difference in age and education background between the two groups. The communication theory assessment score of the experimental group was 84.10±5.27, significantly higher than the control group (72.23±4.48)(P＜0.05). In the experimental group, the score of medical satisfaction after Q-TRUST training was 97.29±1.52, and the number of medical complaints was 15.In the control group. the score of medical satisfaction was 90.75±1.56 and 24 cases of medical complaints were obtained after traditional training. The experimental group was significantly better than the control group(P＜0.05). In the experimental group,the average score of the overall training effect of was 9.43±0.59, the degree of conformity with demand was 9.35±0.80.The results showed that the students in the experimental group were more satisfied with Q-TRUST training course and had high demand evaluation. How to quickly establish trust was 9.72±0.45; how to tell bad news 9.58±0.50; how to improve the efficiency of conversation 9.55±0.22; early identification of high-risk communication patients 9.47±0.87. In the feedback of experimental group, 100% of trainees believed that their communication skills was improved, 93.3% of dispute resolution ability was improved, 90% of communication efficiency and trust building were improved, 86.7% of identification for high-risk patients were improved and 83.3% of etiquette was improved. Conclusions Q-TRUST clinical communication training course can quickly improve the communication ability of new staff, enhance confidence, and better adapt to clinical works of emergency department.

Application of the combination of recorded lectures and online question and answer in postgraduate course of medical imaging

DING Ning, ZHANG Da-ming, SU Tong, XU Jia, JIA Xue-yan, XUE Hua-dan, JIN Zheng-yu

2022, 42(6):  999-1003.  doi:10.16352/j.issn.1001-6325.2022.06.015

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Objective To prospectively design and followed by evaluation on the effect of the combination of recorded lectures with online question and answer(Q & A) in graduate course of medical imaging, so as to provide more experience for future online medical imaging courses. Methods A total of 19 students recruited in the 2020 spring semester course were included. An online questionnaire was used to collect the results of self-assessment on knowledge and skill learning before and after each lecture, participation rate of Q & A activity. Results The knowledge and skills mastery level after learning every 16 lectures was significantly higher than that before the lecture. The average score of students in the Objective test was 44.2±7.0 and the average score of students' participation in the Q & A session was 3.79±0.92. There is no significant correlation between test scores and the participation in Q & A. The average score of students' evaluation of the necessity about the Q & A activity is 4.47±0.90. The average score of students' subjective feelings of traditional classrooms is higher than that of remote virtual classrooms, but the difference is not statistically. Conclusions Recorded lectures plus online Q & A in graduate course of medical imaging has significantly improved students' learning outcomes on knowledge and skill. Online Q & A following the course can increase students' participation and enhance their learning motivation.

Synthetic ability cultivating in clinical-type medical graduates under standardized resident training system

ZHOU Meng-xin, CHEN Yue-xin, YANG Bo

2022, 42(6):  1004-1006.  doi:10.16352/j.issn.1001-6325.2022.06.018

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Merging the education model of clinical-type medical graduates into a double-track system, medical colleges and teaching hospitals are facing challenges of more requirements on cultivating graduates. In order to help the graduates to join in standardized resident training in an active attitude in order to strengthen their capacity building for learning and scientific research, this article discussed the development of integrated capacity during the training of clinical-type medical graduates from three aspects as clinical capacity, innovation spirit and personal quality.

Difficulty and adjustment in clinical teaching of nuclear medicine during COVID-19 pandemic

DAI Yu-hui, CHEN Yong-hui, JING Hong-li, HUO Li

2022, 42(6):  1007-1009.  doi:10.16352/j.issn.1001-6325.2022.06.017

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During COVID-19 pandemic and due to the need for its prevention and control, the training of interns and graduates in nuclear medicine are facing challenges. This paper analyzed these challenges and reported a series of reformation implemented by the Department of Nuclear Medicine of Peking Union Medical College Hospital. During the pandemic, students face various difficulties in the process of clinical practice, which also leads to some psychological problems such as fear, anxiety and depression. Therefore, the department of nuclear medicine remodels the teaching and training plan based on the needs of trainees and constantly modify the training procedures to protect trainees and meet the needs of training as well as pandemic prevention and control.

Medical Management

Significance of practicing “The three most valuable things in the future” for the physical and mental health of the elderly

LI Bin, LIU Ting-ting, HE Li

2022, 42(6):  1010-1012.  doi:10.16352/j.issn.1001-6325.2022.06.013

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“The three most valuable things in the future” are the health-related issues proposed by city government of Beijing in 2021: the first one is defined as “I am healthy, so I will not entangle on my family”; the second is “ I can make others healthy, so to realize the value of life”; the third thing is “I make contribution to social health to make my life meaningful”. Some elder people who we are familiar with have done these very well. When help others in need, they feel very satisfied from being their friend indeed. It is desirable for the elderly to keep a healthy state after retirement. Moreover, they help others to live in health. This spirit comes from their appreciation to the current society environment and great future.