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Table of Content

    05 May 2022, Volume 42 Issue 5
    Original Articles
    Effects of carboxyamidotriazole-orotate on oncogenic phenotypes of mouse glioma-associated macrophage cell lines
    MA Rui, YANG Li-xing, CHEN Qiu-xia, QIU Jia-xing, WANG Yu-cheng, JU Rui, GUO Lei
    2022, 42(5):  701-707.  doi:10.16352/j.issn.1001-6325.2022.05.028
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    Objective To explore regulation mechanism of carboxyamidotriazole-orotate (CTO) on oncogenic phenotype of mouse glioma-associated macrophages. Methods Bone marrow-derived macrophages(BMDMs) or RAW264.7 macrophages (tumor associated macrophages,TAMs) were prepared by super-culture of mouse glioma GL261 cells. The mRNA of TAMs oncogenic mediators was detected by qPCR. The oxygen consumption rate (OCR) of TAMs was measured by Seahorse bioenergy method. The protein level of hypoxia-inducible factor-1 α(HIF-1α), hypoxia-inducible factor-2 α (HIF-2α) and peroxisome proliferator-activated receptor (PPAR) co-activator factor-1 β (PGC-1β) in TAMs was detected by Western blot. Results CTO decreased M1-related mediators such as IL-1β, IL-6 and TNF-α(P<0.001) and M2-related mediators such as IL-10, ARG-1 and TGF-β1(P<0.001) in TAMs. CTO significantly down-regulated OCR in TAMs (P<0.01); The protein level of HIF-1α and of HIF-2α in TAMs was down-regulated by CTO(P<0.01),which was contrary to the effect of carboxycyanide-4-trifluormethoxyphenylhydrazone (FCCP),which in turn promoted mitochondrial oxygen consumption. CTO significantly reduced PGC-1β protein in TAMs (P<0.001),an effect consistent with rotenone(RTN), a mitochondrial respiratory chain inhibitor. Conclusions A decreased oxidative phosphorylation down-regulates the expression of PGC-1β and destabilizes HIF-1α and HIF-2α, thus affects the production of several M1 and M2 phenotypes of carcinogenic mediators in TAMs, which may be a potential mechanism through which CTO improves the chemotherapy efficacy of glioblastoma.
    Association of ABCB1, ABCC4 and SLCO1B1 polymorphisms with high dose-MTX blood concentration and adverse events in childhood acute lymphoblastic leukemia
    TIAN Xiao-yi, LIU Ying, YAO Yao, GU Xiu-li, LI Ai-hua, ZHENG Hu-yong, SONG Wen-qi
    2022, 42(5):  708-713.  doi:10.16352/j.issn.1001-6325.2022.05.029
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    Objective To investigate potential relationship between gene polymorphisms of ATP-binding cassette B1, C4 (ABCB1, ABCC4) and solute carrier organic anion transporter 1B1 gene(SLCO1B1) with methotrexate (MTX) blood drug concentration and high dose-MTX (HD-MTX)-related adverse events (AEs) in childhood acute lymphoblastic leukemia (ALL). Methods From January 2018 to December 2019, the blood samples were randomly collected from 275 hospitalized pediatric ALL patients in Beijing Children's Hospital of Capital Medical University. All patients were treated with HD-MTX. Chemiluminescent microparticle immunoassay (CMIA) was used to determine the MTX blood drug concentration at 45 h after infusion. Genotyping of ABCB1(rs1045642), ABCC4(rs7317112), and SLCO1B1(rs11045879 and rs10841753) were determined using MALDI-TOF MS based on MELPA technology. At the same time, chemotherapy-induced toxicity data including liver dysfunction, gastrointestinal disorder, mucositis, myelosuppression, and myocardial damage were recorded. The relationship of SNP polymorphism of these genes and MTX blood drug concentration with AEs were analyzed. Results All the genotypes analyzed were in Hardy-Weinberg equilibrium(P>0.05). A statistically significant relationship between ABCC4 rs7317112 and SLCO1B1 rs11045879 genotype and 45 h MTX blood drug concentration was found(P<0.05). The ABCC4 rs7317112 AA and AG genotypes and SLCO1B1 rs11045879 TT and TC genotypes may also be potential indicators for the excretion delay of MTX(P<0.05). Conclusions ABCC4 rs7317112 and SLCO1B1 rs11045879 polymorphisms are potential genetic markers for individual MTX doses in the treatment of pediatric ALL patients.
    Establishment and comparison of in vitro blood-brain barrier cell models
    SU Yu-wen, XIU Jian-bo, XU Qi
    2022, 42(5):  714-720.  doi:10.16352/j.issn.1001-6325.2022.05.027
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    Objective To construct and evaluate the in vitro models of blood-brain barrier (BBB)and to analyze their structural and functional characteristics. Methods Human microvascular endothelial cell line(hCMEC/D3), Uppsala 87 malignant glioma cell line(U87MG) and human brain vascular pericyte(HBVP) cell line were used to construct blood-brain barrier cell models. The models were divided into single-culture model, bi-culture (hCMEC/D3+U87MG or HBVP), no-contact co-culture model, semi-contact co-culture and full-contact co-culture model according to different cell types and cell locations. The permeability and tightness of the models as well as the expression of related proteins and genes were detected for comparing and analyzing the structural and functional characteristics of different models. Results Compared with the single-culture model and the bi-culture model, the permeability and compactness of the co-culture model decreased and the expression of BBB-related genes and proteins also increased(P<0.05). In the co-culture models, full-contact co-culture model showed higher compactness(P<0.01) and higher BBB-related protein and gene expression. Semi-contact and full-contact culture model had lower permeability(P<0.05). Conclusions Full-contact co-culture model has better BBB-related properties and is more suitable for BBB research.
    Expression of CBX7 protein in cervical cancer and its clinical significance
    Hatimihan· MAIMAITI, Gulikezi·MAIMAITIREXIATI, LI Yu-ting, ZHOU Chang-hui, LI Rong
    2022, 42(5):  721-725.  doi:10.16352/j.issn.1001-6325.2022.05.006
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    Objective To investigate the expression and clinical significance of chomobox homolog 7 (CBX7) in cervical cancer(CC)tissues. Methods Tissue samples of 71 patients with CC who were first diagnosed with CC and underwent cervical resection in the First Affiliated Hospital of Xinjiang Medical University from 2013 to 2020 were retrospectively collected. The expression of CBX7 was detected by immunohistochemistry(IHC). At the same time, data of clinical pathology from the patients were collected and survival analysis was conducted after following-up. Results CBX7 was highly expressed in 10 CC tissues, moderately low in 19 CC tissues, and negative in 42 CC tissues(P<0.05). The expression of CBX7 was different and rlated to clinical stages, lymphatic metastasis and vascular invasion(P<0.05). Compared with the negative and low CBX7 expression groups, the survival rate of CC patients with high CBX7 expression was significantly enhanced (P<0.05)and the survival rate of patients with different CBX7 expression levels were significantly different (P<0.05). Conclusions The negative expression of CBX7 protein in CC tissues may be associated with poor prognosis of CC patients.CBX7 expression in CC tissues is a potential prognostic fact of CC patients.
    Polyclonal antibody preparation and identification of rabbit anti-mouse MORF4L1 isoform a
    LI Fang, ZHANG Yuan, ZHANG Ye
    2022, 42(5):  726-731.  doi:10.16352/j.issn.1001-6325.2022.05.025
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    Objective To prepare rabbit anti-mouse polyclonal antibody against MORF4L1 isoform to differentiate MORF4L1 isoform a and isoform b. Methods According to the antigenicity and autoploidy analysis of amino acid sequence of mouse MORF4L1 isoform a and isoform b,the N-terminal 52-66 amino acids peptide sequence (a 15 peptide,KSAVRPRRSEKSLKT) of mouse MORF4L1 isoform a was synthesized. Then the peptide was linked to a carrier protein KLH (keyhole limpet hemocyanin) as an antigen to prepare the antibody against MORF4L1 isoform a. The antiserum of rabbit 1# with good specificity was purified by antigen affinity chromatography and the antibody was titered by ELISA. In addition, the prokaryotic and eukaryotic expression plasmids of MORF4L1 isoform a and b were constructed. Western blot was used to identify the specificity of the antibody by using the MORF4L1 isoform a and b. Results The antiserum of rabbit 1# recognized polypeptide specifically. The titer of purified antibodywas 1:512 000.pGEX-4T-2-M4S,pGEX-4T-2-M4L prokaryotic expression plasmids and pCMV-3Tag-6-M4S,pCMV-3Tag-6-M4L eukaryotic expression plasmids were successfully constructed and expressed by prokaryotic and eukaryotic expression system. The antibody binds specifically to MORF4L1 isoform a. Conclusions Rabbit anti-mouse MORF4L1 a antibody with high titer and high specificity is successfully prepared, which lay an experimental foundation for further revealing the biological significance of MORF4L1 alternative splicing and the biological functions of different MORF4L1 isoforms.
    Cloning, expression and identification of human ATP11A and ATP11B
    HAN Tian-tian, ZENG Jing, YIN Jie, LI Xiao-ying, JIN Ying, LI Yan, PAN Ling-ya
    2022, 42(5):  732-739.  doi:10.16352/j.issn.1001-6325.2022.05.023
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    Objective To construct the expression vectors carrying human Type Ⅳ-P-type phospholipids transferases 11A or 11B (ATP11A or ATP11B), and to detect the up-regulation effect on ATP11A or ATP11B. Methods The primers were designed according to ATP11A or ATP11B sequence and cloned into EZ-TTM Cloning Vector. The primers were identified by enzyme digestion and verified by sequencing. ATP11A and ATP11B sequences were digested and directed to expression vectors pLVX-IRES-mCherry and pLVX-IRES-RFP, respectively. Then the recombinant expression plasmids with immunofluorescence protein were transfected into HEK-293T using liposomes. Finally, the expression of the recombinant cell strains was verified by sequencing, fluorescence microscopy, flow cytometry and Western blot. Results The construction of ATP11A and ATP11B cloning vectors was confirmed by gel electrophoresis and was analyzed by endonuclease. The efficiency of transfection was observed qualitatively under microscope, detected quantitatively by flow cytometry as 38.9%,41.9% respectively, and the tagged proteins were detected by Western blot to present only in the positive groups. Conclusions The expression vectors of human ATP11A and ATP11B are successfully constructed and their expression levels could be effec-tively upregulated,which lay a foundation for future mechanism studies involved to ATP11A or ATP11B.
    Expression of Fc-gamma receptor type Ⅰ in brain cortex of patients with Alzheimer's disease and the mouse model
    SUN Jian-ru, QIU Wen-ying, QIAN Xiao-jing, MA Chao
    2022, 42(5):  740-744.  doi:10.16352/j.issn.1001-6325.2022.05.004
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    Objective To explore expression changes of high affinity IgG receptor Fc-gamma Ⅰ type(FcγRⅠ) in brain cortex of patients with Alzheimer's disease(AD). Methods Normal human brain tissue samples and brain tissue samples with neuropathological diagnosis of AD, wild type mice and APP/PS1 mice were used. Immunofluorescence and Western blot were used to define cellular localization and expression of FcγRⅠ in cerebral cortex. Results FcγRⅠ was generally expressed in neurons of human and mouse cerebral cortex. Compared with normal human brain, FcγRⅠ expression in AD human brain cortex decreased significantly(P<0.05). Compared with wild type mouse brain, FcγRⅠ expression in cerebral cortex of AD mouse models also decreased significantly(P<0.05). Conclusions The expression of IgG high affinity receptor FcγRⅠ in AD human brain and cerebral cortex of AD mouse models decreases significantly that suggestes that FcγRⅠ is potentially correlated with AD neuropathological pathogenesis.
    Angiopoietin-like protein 8 promotes phenotypic transformation of rat aorta vascular smooth muscle cells
    SUN Qiu-ju, JIAO Xiao-lu, YU Hua-hui, LI Fan, LYU Qian-wen, WANG Yu, QIN Yan-wen
    2022, 42(5):  745-751.  doi:10.16352/j.issn.1001-6325.2022.05.009
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    Objective To explore the role and mechanism of angiopoietin-like protein 8 (ANGPTL8) on phenotypic transformation of rat aorta vascular smooth muscle cells (VSMCs). Methods ApoE-/- male mice (8 weeks of age) were fed with a western diet for 12 weeks to build the atherosclerosis mouse model. The expression and localization of ANGPTL8 in atherosclerotic plaques were detected by immunofluorescence staining. Stable transfections of the ANGPTL8 over-expression vector were performed. VSMCs were randomly divided into 5 groups: control group,LV-ANGPTL8 group,ox-LDL (50 mg/L)group,(ox-LDL+LV-ANGPTL8) group and (LV-ANGPTL8+PD98059 group. And cell migration was observed by wound-healing assay. The protein expressions of α-SMA, SM22α, OPN, PCNA and Ki67 in VSMCs was detected by Western blot. Results ANGPTL8 over-expression promoted the expression of OPN, PCNA and Ki67 in VSMCs (P<0.05), decreased the expression of α-SMA and SM22α(P<0.05) and enhanced migration ability of VSMCs(P<0.05). After adding ERK pathway inhibitor PD98059, the expression of OPN, PCNA and Ki67 decreased (P<0.05), α-SMA and SM22α expression increased(P<0.05), and the migration capability of the VSMCs decreased (P<0.05). Conclusions ANGPTL8 maybe promotes phenotypic transformation of VSMCs by activating ERK pathway.
    Screening and identification of broadly neutralizing antibodies against different mutant strains of SARS-CoV2
    TONG Ke-xin, CAI Meng-hua, HE Wei, ZHANG Jian-min
    2022, 42(5):  752-757.  doi:10.16352/j.issn.1001-6325.2022.05.022
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    Objective To screen high-affinity neutralizing antibodies with broad spectrum specificity against different SARS-CoV2 mutants and verify their functions. Methods Balb/c mice were immunized twice with SARS-CoV2 S protein mRNA vaccine coated with liposome, and the specific antibody content in serum was determined. The mice with higher antibody titer were selected for shock immunization. The spleen cells of mice after shock immunization were fused with myeloma cells to prepare fusion cells, and the clones that produces monoclonal antibodies against different parts of S protein were screened by limited dilution and ELISA. Monoclonal antibody was purified and neutral-ization experiments were performed to verify the neutralization activity of the antibody. Monoclonal antibodies with broad spectrum binding properties against recombinant receptor binding domain(RBD) proteins of four SARS-CoV2 mutants were screened by ELISA. Results After twice immunizations, the serum antibody titer against S protein in high dose group 4 was the highest. After shock immunity and cell fusion, 12 hybridoma cell clones with binding activity to S protein were screened, among which 9 strains specifically bound to RBD protein. The neutralization test results showed that the antibodies 41, 46 and 47 had significant neutralization activity in vitro. Compared with the other two antibodies, antibody 46 showed broad-spectrum binding activity to the RBD proteins of four SARS-CoV2 mutants including Omicron. Conclusions Antibody 46 is a broad-spectrum specific neutralizing antibody against four different SARS-CoV2 mutants, which can be further humanized and potentially used for the treatment of COVID-19.
    Transcription factor Kaiso down-regulates the expression of lncRNA MEG3 in human breast cancer cell lines
    XIE Li-hua, YANG Qi, LI Ya-qian, XIE Xin-yue, LEI Shi-juan, ZHOU Lin
    2022, 42(5):  758-762.  doi:10.16352/j.issn.1001-6325.2022.05.024
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    Objective To investigate the potential regualation of the expression of human maternal gene 3(MEG3) by transcription factor Kaiso and the impact on the occurrence and development of breast cancer. Methods The expression of Kaiso and MEG3 in breast cell line MCF-10a and different breast cancer cell lines MCF-7,BT-474,MDA-MB-435,MDA-MB-231 were detected by RT-qPCR. The expression of Kaiso in cells was detected by Western blot.Kaiso expression of 50 samples from cancer and paracancer tissues were examined by Western blot. RT-qPCR was used to detect the effect of Kaiso over-expressed on the transcription of long non-coding RNA(lncRNA) MEG3. Results The transcription of lncRNA MEG3 in MCF-10a was higher than that of Kaiso (P<0.001). Kaiso was significantly up-regulated in MCF-7, BT-474, MDA-MB-435, and MDA-MB-231 breast cancer cells,while lncRNA MEG3 was significantly down-regulated (P<0.001). Kaiso expression level was low in MCF-10a, but was high in MDA-MB-231 highly metastatic breast cancer (P<0.05). Kaiso was low in paracancerous breast tissues (P<0.001), but highly expressed in cytoplasm and nucleus in cancer tissues. In addition, Kaiso over-expression in MCF-10a significantly down regulated the transcription of lncRNA MEG3(P<0.01). Conclusions Kaiso regulates the expression of lncRNA MEG3 by breast cancer cells, which may play an important role in the occurrence and development of breast cancer.
    Chromatography and immunogenicity analysis of HPV-58 L1 virus-like particles expressed from insect cells
    WANG Zhi-rong, ZHANG Ting, XU Xue-mei
    2022, 42(5):  763-767.  doi:10.16352/j.issn.1001-6325.2022.05.018
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    Objective To develop a simple chromatography purification process of human papillomavirus (HPV) type 58 L1 virus-like particle (VLP) produced by insect cells baculovirus expression vector system (IBEVS). Methods A previously constructed mutant of HPV-58 L1 with a high expression level in insect baculovirus system were used to prepare the cell lysates.The L1 containing sample was used for the establishment of optimal protein purification process which consisting of an initial process of ammonium sulfate precipitation (ASP) and two polishing chromatography purifications. Purified L1 protein was dialyzed for VLP reassembly and the resultant VLPs were characterized physicochemically and their immunogenicity was then analyzed in mice. Results L1 protein was purified by 30% ASP with good purity (9.34%) and recovery (72%). When bound to anion exchange resin and eluted by 300 mmol/L NaCl-contained buffer, the L1 protein was further purified with >90% of recovery and 13.6% of purity. The resultant L1 protein mainly flowed through the ceramic hydroxyapatite column with a recovery of >80% and purity was over 99.1% by HCP analysis. With the three-step purification, the total recovery of L1 was about 55.7% and the yield was 57-65 mg/L. Purified L1 proteins self-assembled into homogenous VLPs with an average diameter of 55 nm as proved by TEM and DLS, and these VLPs induced high titer of neutralizing antibodies, comparable to that of VLPs purified by ultracentrifugation. Conclusions The chromatography process for producing HPV-58 L1 VLPs showed good purity, yield, and immunogenicity and is believed to be a time-saving, easy-performing, fully scalable, and low-cost method. It may be used to produce low-cost and multivalent HPV VLP vaccine by IBEVS.
    LncRNA FGD5-AS1 reduces ox-LDL-induced damage of HUVECs by targeting miR-106a-5p
    GUO Jing, LI Ya-jie, MOU Han-shuang
    2022, 42(5):  768-775.  doi:10.16352/j.issn.1001-6325.2022.05.026
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    Objective To explore the effect of lncRNA FGD5-AS1 on oxidized low-density lipoprotein (ox-LDL)-induced vascular endothelial cell damage and its regulatory effect on miR-106a-5p. Methods Ox-LDL was used to induce human umbilical vein endothelial cells(HUVECs) to establish a cellular injury model. pcDNA-FGD5-AS1, anti-miR-106a-5p and its negative control were transfected into HUVECs and then added to ox-LDL-treated cells. pcDNA-FGD5-AS1 was co-transfected with miR-NC and miR-106a-5p mimics to HUVECs, then added ox-LDL to treat cells. RT-qPCR was used to detect the expression of FGD5-AS1 and miR-106a-5p. The level of MDA, SOD, and CAT was tested with commercially available kit. Flow cytometry was used to detect the apoptosis rate. The dual luciferase reporter gene experiment was used to detect the targeting relationship between FGD5-AS1 and miR-106a-5p. Western blot was used to detect the protein expression of cleaved caspase-3 and cleaved caspase-9. Results The expression of FGD5-AS1 mRNA in HUVECs induced by ox-LDL decreased (P<0.05) while the expression of miR-106a-5p mRNA increased (P<0.05). Both transfection of pcDNA-FGD5-AS1 and transfection of anti-miR-106a-5p reduced the level of MDA, apoptosis rate and protein level of cleaved caspase-3, cleaved caspase-9 in HUVECs induced by ox-LDL(P<0.05),enhanced the activity of SOD and CAT (P<0.05). FGD5-AS1 could target at miR-106a-5p. Co-transfection of pcDNA-FGD5-AS1 and miR-106a-5p mimics could reverse the effect of pcDNA-FGD5-AS1 on ox-LDL-induced HUVECs apoptosis and oxidative stress. Conclusions Over-expression of FGD5-AS1 may inhibit oxidative stress and apoptosis through targeted regulation of miR-106a-5p, thereby reducing ox-LDL-induced vascular endothelial cell damage.
    Single-cell RNA sequencing based analysis depicts the development map of human early embryonic erythrocytes
    CHEN Zhong-yang, MA Yan-ni, YU Jia
    2022, 42(5):  776-781.  doi:10.16352/j.issn.1001-6325.2022.05.019
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    Objective Single-cell RNA sequencing (scRNA-seq) was used to systematically delineate the cell atlas and molecular tracks of human early erythroid development at the single-cell level. Methods The dataset of scRNA-seq of human embryo yolk sac at CS15 stage was analyzed by principal component analysis (PCA), cluster analysis and pseudostime analysis to observe the expression of marker genes in different cell clusters, and to predict the occurrence time and development path of erythroid. Results In different cell clusters of yolk sac at CS15 stage, marker genes of erythrocytes and progenitor cells were differentially expressed. The occurrence time of various cells was different, and there was some heterogeneity in erythrocytes in the yolk sac. There were abundant types of cells in the yolk sac at CS15 stage, which could include vascular endothelial cells, hematopoietic progenitor cells, granulocytes, different status of red blood cells and monocytes. Conclusions It is speculated that early development path of human erythroid is as follows: vascular endothelial cells differentiate into hematopoietic progenitor cells, further differentiate into red myeloid progenitor cells, and finally produce megakaryocytes and erythrocytes.
    Establishment and optimization of air-liquid interface method cultured mouse-derived tumor organoids
    LI Zhao-xuan, LUO Yun-ping, CHEN Chong
    2022, 42(5):  782-787.  doi:10.16352/j.issn.1001-6325.2022.05.011
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    Objective To construct a mouse-derived tumor organoid that can retain various types of immune cells in the immune microenvironment without artificial reconstruction. Methods The mouse-derived tumor organoids were cultured and constructed by air-liquid interface method. The histological morphology of the tumor organoids was analyzed by hematoxylin and eosin(HE). The proportion of various types of immune cells in the tumor organoids was analyzed by flow cytometry. The interleukin-2 (IL-2) was used to maintain the number of T lymphocytes in tumor organoids. Results The tumor organoids from three tumor types of mouse CT26 colon cancer, Lewis lung cancer and MC38 colon cancer all retained the histomorphology of the original tumor. Compared with mouse CT26 colon cancer tumor tissues, the proportion of CD4+T cells, CD8+ T cells and B cells in tumor organoids didn't show significant change and the proportion of macrophages decreased from 66.7% to 44.3%. The proportion of dendritic cells decreased from 66.5% to 7.25%. Compared with mouse Lewis lung cancer tumor tissue, the proportion of CD4+T cells, macrophages, dendritic cells and B cells in tumor organoids remained unchanged and the proportion of CD8+ T cells decreased from 5.6% to 1.95%. Compared with mouse MC38 colon cancer tumor tissue, the proportion of CD4+T cells, CD8+ T cells and B cells in tumor organoids remained unchanged, the proportion of macrophages decreased from 33.5% to 17%, and the proportion of dendritic cells decreased from 46.8% to 1.2%. IL-2 could increase the tumor-infiltrating T lymphocytes in tumor organoids derived from three tumor types. Conclusions Mouse tumor organoids retain the histological morphology of the original tumor and the various types of immune cells are retained in a certain extent. IL-2 can increase the quantity of T lymphocytes.
    Effect of obesity and obesity trajectories on the risk of cardiovascular diseases in adult male with incident NAFLD in Northern China
    HU Shi-qi, LAN Yan-qi, WU Shou-ling, WANG Xiao-mo, CHEN Shuo-hua, WANG Guo-dong, WANG Li
    2022, 42(5):  788-794.  doi:10.16352/j.issn.1001-6325.2022.05.021
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    Objective To investigate the baseline and effect of obesity and obesity trajectories on cardiovascular diseases(CVDs) in adult male with incident non-alcoholic fatty liver disease(NAFLD). Methods Adult men with newly diagnosed NAFLD and without CVDs in the Kailuan cohort study from 2006—2013 were enrolled. The subjects were divided into the nonobese group and obese group according to the definition of body mass index(BMI) or waist circumference (WC), then further divided by the obesity trajectories at the first follow-up (1-4 years). Kaplan-Meier method was used to estimate the incidence rate of CVDs for each subgroup. In addition, the Fine and Gray competing risk Cox regression model was used to compare CVDs risk in NAFLD with different baseline or trajectories in obesity measures after 1-4 years of NAFLD diagnosis. Results A total of 8 591 incident NAFLD patients were included. During a median follow-up of 9.41 years, 349 cases were found with CVDs. The 10-year cumulative incidence rates of CVDs were 3.3% and 4.3% in the nonobese and obese groups according to the definition of waist circumference, respectively (log-rank test, P<0.05). However, the significance disappeared after adjusting the competing risk of death and other covariates. Similar results were observed based on the definition of BMI. Also, further analysis was conducted for the association trajectories in obesity measures and CVDs. The 8-year cumulative incidence rates of CVDs were 2.8%, 3.6%, 4.2%, and 3.3% in the constantly nonobese group, nonobese-obese, obese-nonobese, and constantly obese group, respectively (log-rank test, P>0.05). Compared to the subjects with constantly nonobese defined by BMI, those who transition from nonobese to obese increased the CVDs risk by 57% (hazard ratio [HR]:1.57; 95% CI:1.03~2.38; P<0.05). The conclusion remained even after adjusting baseline BMI (HR:1.56; 95% CI:1.02~2.37; P<0.05). Conclusions Short-term weight gain in men with incident NAFLD led a higher CVDs risk, suggesting the importance of weight control in preventing CVDs once NAFLD occurrence.
    Clinical Sciences
    Analysis of related factors influencing the selection of intubation methods under general anesthesia for patients with ankylosing spondylitis
    ZHANG Yu-guan, XIAO Bo, ZHANG Yue-lun, YI Jie
    2022, 42(5):  795-798.  doi:10.16352/j.issn.1001-6325.2022.05.007
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    Objective To investigate the degree of difficult intubations for patients with ankylosing spondylitis (AS) and to predict the related factors influencing the selection of intubation methods. Methods A total of 347 patients(276 males, 71 females), aged 15-77 years, ASA Ⅰ-Ⅲwith AS underwent surgical procedure from January 2008 to January 2017 were included. During general anesthesia induction, patients were divided into two groups according to the application of non-visual devices (179 cases) and visual devices (168 cases). Patient data were collected, including gender, age, kyphosis, degree of mouth opening, Mallampatti classification, neck mobility and the duration of the AS. Multivariate regression analysis of variables was conducted to find the relevant predictors of intubations method selection in AS patients with difficult endotracheal airway. Results The older patients, kyphosis, limited mouth opening, limited neck range of motion, and longer duration of AS, the utilization rate of the visual group was significantly higher than that of the non-visual group (P<0.05). Multi-factor Logistic regression analysis suggested that moderate limitation of neck range of motion(OR=5.41,95% CI: 2.32-13.10,P<0.001)and severe limitation of neck range of motion (OR=9.22,95% CI: 4.37-20.25,P<0.001) were related factors for tracheal intubation with visual devices in patients with difficult airway. Conclusions Neck mobility has a great influence on whether or not to use visual intubation tools. When neck mobility decreases, anesthesiologists are more likely to use visual devices such as video laryngoscope, McCoy laryngoscope, visual rigid and fiberoptic bronchoscope.
    Mini Reviews
    Advances in research on relationship of circadian disruption with cardiac dysfunction
    LIANG Qing, CHEN Li-hong
    2022, 42(5):  799-803.  doi:10.16352/j.issn.1001-6325.2022.05.005
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    Accumulating evidence has shown that disruption of the circadian timing system by either endogenous genetic variants or exogenous environmental factors is significantly associated with increased risk of cardiovascular diseases, such as myocardial infarction, arrhythmia, sudden cardiac death and so on. It is of great significance to understanding the relationship between circadian disruption and cardiovascular function and disease.
    Application of anti-HBc quantitative examination in clinical management of chronic hepatitis B virus infection
    DU Xin-yi, MAO Xiao-rong, LI Jun-feng
    2022, 42(5):  804-808.  doi:10.16352/j.issn.1001-6325.2022.05.010
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    The quantification of hepatitis B virus(HBV) core antibody (anti-HBc) is related to host immunity and can be detected in the serum of patients with current or past infections. The antibody against core antigen of HBV plays an important role in the inflammation pathogenesis of liver,evaluation of epidemiology history and of treatment strategy. Chronic HBV infection is closely related to active transcription and persistence of covalently closed circular DNA(cccDNA) in the liver. The result of quantitative anti-HBc antibody examination shows a good correlation with cccDNA level so it is a potential index for comprehensive evaluation of HBV infectious diseases.
    Research progress on the inflammatory mechanisms of cholestatic liver injury
    KANG Jin-yu, LUO Jia, GAO Ze-tian, ZHENG Wen-tao, LIU Ai-ming, SONG Yu-fei
    2022, 42(5):  809-813.  doi:10.16352/j.issn.1001-6325.2022.05.002
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    Cholestatic liver injury (CLI) is a liver disease with high mordity. Accumulation of bile acids in the liver is thought to be a major driver of liver injury during cholestasis. The disease can develop into be liver fibrosis, cirrhosis, liver failure and liver cancer. This review focuses on the molecular mechanism of CLI in the perspective of inflammatory response. The roles of neutrophils, Kupffer cells, bile duct cells, and the signaling pathways involved in CLI are reviewed to provide clues for research of new therapeutic targets and treatment strategies.
    Progress on the pathogenesis of cardiovascular complications in patients with type 2 diabetes mellitus
    ZHENG Tian-sheng, TONG Xue-wei, ZHANG Yi-tong, ZHU Min, ZHAN Xiao-rong
    2022, 42(5):  814-817.  doi:10.16352/j.issn.1001-6325.2022.05.008
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    Patients with type 2 diabetes mellitus(T2DM) trend toward the multiple cardiovascular complications like coronary atherosclerotic heart disease, hypertension, heart failure. The causes and mechanisms leading to cardiovascular disease in patients with type 2 diabetes mellitus are complicated, which include persistent hyperglycemia, insulin resistance, metabolic disorders, oxidative stress, myocardial fibrosis, small vessel disease and cardiac autonomic neuropathy.
    Iron metabolism in human body and its regulatory factors
    CHEN Chun-mei, GE Pin, GUO Chong
    2022, 42(5):  818-823.  doi:10.16352/j.issn.1001-6325.2022.05.017
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    This article reviews the molecular mechanism of iron metabolism covering iron absorption,transportation,utilization and storage with focus on the key proteins involved in iron metabolism and elaborate regulation factors. Based on these results of research it might be able to provide some new strategies to find potential biomarkers of iron metabolism to support early and correct diagnosis. What's more, a clear mechanism may also provide a breakthrough point for the targeted treatment of iron metabolism disorders in future.
    Application of anti-thrombotic drugs and the safety of prostate biopsy
    ZUO Yu-zhi, YAN Wei-gang
    2022, 42(5):  824-827.  doi:10.16352/j.issn.1001-6325.2022.05.003
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    Incidence rate of prostate cancer has been ranked as the first one in male malignant tumors. Prostate biopsy is the golden standard for diagnosis of prostate cancer. However, clinical treatment of prostate cancer is facing the challenge of lacking evidence-based recommendations for peri-operative anti-thrombotic medication management are lacking. Through reviewing of research papers, it was found that administration of oral anti-thrombosis drugs was safe for patients undergoing prostate biopsy. For patients with high risk of cerebro-cardiovascular diseases. Continuing use of anti-thrombotic drugs during peri-operative period is suggested.
    Medical Education
    Exploration of case-based teaching in endocrinology and metabolism continuing medical education
    ZHANG Qian, XIAO Xin-hua
    2022, 42(5):  828-831.  doi:10.16352/j.issn.1001-6325.2022.05.001
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    As an efficient teaching model, case-based teaching (CBT) takes the case as a center issue and the teacher introduces the classic case in the teaching process and explains it in combination with the theory. This method is conducive to deepen the students' understanding of the pathological knowledge and stimulates students' learning enthusiasm with clear targets. Endocrinology and metabolism are important branches of clinical medicine. With the rapid development of knowledge in the field of endocrinology and metabolism, the demand of continuing medical education is growing. The traditional course of continuing medical education is based on a one-way teaching mode. The application of CBT can be inspired and guided by teachers. Through classroom discussions and other forms peer education, it helps the student to better understand medical knowledge . At the same time, it also helps to broaden students' knowledge, improve their capacity building on analysis, problem solving,critical thinking, innovation and self-regulated learning. The application of CBT in continuing teaching of endocrinology and metabolism can significantly improve the teaching quality and learning outcomes of the trainees with more academic resources.
    Impact of trainer experience and student category on quantitative evaluation for surgical teaching in the department of breast surgery
    WANG Chang-jun, LIN Yan, ZHOU Yi-dong, MAO Feng, SHEN Song-jie, SUN Qiang
    2022, 42(5):  832-835.  doi:10.16352/j.issn.1001-6325.2022.05.012
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    Objective To investigate the impact of trainer experience and the types of trainees on quantitative teaching score for surgical education. Methods The present study adopted two-way repeated measures design. The quantitative teaching scores were collected between September 2018 and January 2020. The impact factors were trainer experience and student category. Repeated measure was performed by recording teaching score each month. Results The average of overall teaching score was 0.98±1.58/Teacher/Month. Both teacher experience(P<0.001) and student category (P<0.001) had significant impact on teaching score, and these two factors had interaction effect. Teacher experience had remarkable influence on resident and graduate subgroup compared to the other student category. In resident and graduate subgroup, trainer experience had a positive correlation with teaching score (All the between-subgroup comparisons had P<0.001). However, this effect diminished in the other student category. Conclusions The present teaching system puts in great effort to surgical education for resident and graduate students. And trainer experience is positively correlated with quantitative teaching score.
    Application of electronic prescriptions for narcotic drugs in quality control
    BAI Bing, YANG Gen-zhi, ZHANG Yue-lun, TIAN Yuan, SHEN Le, HUANG Yu-guang
    2022, 42(5):  836-839.  doi:10.16352/j.issn.1001-6325.2022.05.013
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    Objective To investigate the significance of electronic prescriptions on the management of narcotic drugs, so as to constantly optimize the management of narcotic drugs. Methods Continuous random checks were conducted on the prescriptions of narcotic drugs in operating room of the department of anesthesiology of Peking Union Medical College Hospital from March 2019 to December 2021. The changes of the error rate of narcotic drugs prescriptions before and after the electronic prescriptions of narcotic drugs were officially put into use were compared. Results The error rate of narcotic drugs prescriptions before and after electronic prescriptions were 31.5% and 8.0%[odds ratio (OR): 0.188, 95% confidence interval (CI): 0.164-0.215, P<0.001]. Conclusions Electronic prescriptions of narcotic drugs may help to reduce the occurrence of error from manual prescription and has potential significance in optimizing the management of narcotic drugs.
    Investigation on the gynecological endocrinology teaching methods for eight-year program of clinical medicine
    WANG Han-bi, WANG Tao, DENG Cheng-yan
    2022, 42(5):  840-843.  doi:10.16352/j.issn.1001-6325.2022.05.014
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    Objective To investigate the gynecological endocrinology teaching demands and practical effects of rotation for eight-year medical students at Peking Union Medical College(PUMC). On this basis, according to the feedback of the students and the characteristics of gynecological endocrine diseases, the teaching characteristics and the best way of gynecological endocrinology were discussed. Methods The questionnaire survey was issued to the eight-year medical students from September 1, 2020 to December 31, 2020, which investigated the demands of the gynecological endocrine study, the difficulty in learning and the degree of mastery. Results A total of 80 valid questionnaires were received, which showed that the eight-year medical students had a most eager to learn content of the top 3 diseases in turn was polycystic ovary syndrome, uterine bleeding and menopausal syndrome. The top 3 gynecological endocrine diseases with the great difficulty in clinical learning were in turn abnormal sexual development, abnormal uterine bleeding, and recurrent abortion. Both the outpatient and ward teaching were demanded. The most desirable mode of study was outpatient visits. Conclusions Gynecological endocrinology is a unique teaching content of obstetrics and gynecology. The outpatient teaching should be paid more attention. Reasonable arrangement of class hours should be combined with social needs and the difficulty of learning content. In teaching of gynecological endocrine, standardized patients(SP) combining case method of instruction should be effectively achieve ideal teaching results.
    Evaluation of the Jigsaw method in the training of prostate targeted biopsy
    ZHOU Zhi-en, YAN Wei-gang, ZHOU Yi, ZUO Yu-zhi, LIANG Zhen
    2022, 42(5):  844-847.  doi:10.16352/j.issn.1001-6325.2022.05.015
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    Objective To evaluate the effect of the Jigsaw method in training of prostate targeted biopsy. Methods Thirty urologists who participated in the prostate biopsy training course in Peking Union Medical College Hospital in December 2020 were selected as the objects of study. They were randomly divided into two groups. The control group was trained with traditional teaching method and the observation group was trained by a Jigsaw method. Both groups of members were tested by same standard on theory and operation skills and surveyed for their by satisfaction by questionnaires. Results The results of examination of theory and operation skill in the observation group were significantly higher than those in the control group(P<0.05). The questionnaire survey showed that the trainees in observation group were significantly more satisfied with the method than those in the control group(P<0.05). Conclusions The Jigsaw method applied in the training of prostate targeted biopsy can stimulate trainees'learning enthusiasm and get remarkable learning outcomes so is worthy to be popularized.
    Establishment of Huang Jiasi Memorial Hall using of alumni resources is a new mode of innovation and exploration of alumni work
    HUANG Fu-min, LIU Wen-hao, LIU Jing
    2022, 42(5):  848-851.  doi:10.16352/j.issn.1001-6325.2022.05.016
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    As the“brand image”and “talent product”, each member of alumni functions as the business card of the school in terms of development and core capacity competitiveness. Alumni play an irreplaceable role in the enhancement of school reputation, cohesion, inheriting network and accelerating development. Taking the establishment of Huang Jiasi Memorial Hall launched by the alumni of Peking Union Medical College as an example, this paper is of positive significance to the innovative work of the Alumni Association of Peking Union Medical College and to explore the innovative and effective ways for capacity building for alumni performance.
    Management of collection and characteristics of scientific data generated from basic medicine
    WANG Xue-yan, JING Yan-ling, ZHAO Ai-fang
    2022, 42(5):  852-856.  doi:10.16352/j.issn.1001-6325.2022.05.020
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    The article summarizes the collection and management of data from scientific research and introduced technical details. Analysis covers the general characteristics of 29 projects from studies of basic medicine in 2020, which shed light on overall understanding of the scientific data collection process and its implementation. In addition, this study substantially facilitates capacity building for gathering, processing and managing of research data while promoting open sharing. circulation, utilization of data and yielding added-value.