Table of Content

05 March 2022, Volume 42 Issue 3

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Invited Reviews:Atherosclerosis:Focusing on the Plaque Stability

Assessment and targeting in the stability of atherosclerosis plaques

ZHANG Wei-li

2022, 42(3):  357-359.  doi:10.16352/j.issn.1001-6325.2022.03.001

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Application of intravascular optical coherence tomography in detection of coronary atherosclerotic plaque

QIU Yao-yang, GUI Jia-hui, HUANG Lin, HU Xue-qiang, LI Qin

2022, 42(3):  360-365.  doi:10.16352/j.issn.1001-6325.2022.03.002

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Intravascular optical coherence tomography (IVOCT) technology has developed very rapidly in the detection of coronary atherosclerotic plaque in recent years. It can realize high-resolution, radiation-free and real-time in vivo imaging of intravascular diseased tissue, and plays an important role in clinical diagnosis. This paper briefly introduces the imaging principle of IVOCT, calculation methods of the optical characteristic parameters of biological tissue and the dispersion coefficient. The plaque recognition algorithm based on machine learning is also briefly summarized, so as to provide a new strategy and pathway for the development of constructing the artificial intelligent recognition system of plaque.

Effects of lipid-lowering therapy on coronary atherosclerotic plaque evaluated by intravascular imaging

ZHANG Shuang, LI Zhi-fan, QIAN Jie, WU Na-qiong

2022, 42(3):  366-371.  doi:10.16352/j.issn.1001-6325.2022.03.032

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The progression of atherosclerotic plaque is one of the most important causes of cardiovascular events in patients with atherosclerotic cardiovascular diseases (ASCVD). With the help of intravascular imaging, such as intravascular ultrasound (IVUS), optical coherence tomography (OCT) and near-infrared spectroscopy (NIRS), it is possible to show the details of the plaque morphological characteristics. Current research suggested that lipid-lowering drugs lead to plaque stabilization and regression mainly by reducing low-density lipoprotein cholesterol (LDL-C). This review focuses on the effects of lipid-lowering therapy on coronary atherosclerotic plaque evaluated by intravascular imaging.

The role of vascular aging in atherosclerosis

ZHANG Qi-yue, ZHANG Wei-li

2022, 42(3):  372-377.  doi:10.16352/j.issn.1001-6325.2022.03.033

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Vascular aging is an age-dependent degeneration of vascular structure and function, which can lead to an increased risk of cardiovascular diseases. Atherosclerosis is an essential pathogenesis of cardiovascular diseases. Vascular aging plays an important role in the progression of atherosclerosis. Effective evaluation of vascular aging is necessary for the clinical treatment of atherosclerotic diseases. This review summarized the structural and functional changes of vascular aging, and highlighted the important roles of telomere attrition, mitochondrial dysfunction and inflammation of vascular cells in the process of atherosclerosis, aiming to provide new impetus for the clinical treatment of atherosclerotic diseases.

Shear-sensitive factors in therapeutic targets for atherosclerosis

ZHAO Chuan-rong, ZHOU Jing

2022, 42(3):  378-383.  doi:10.16352/j.issn.1001-6325.2022.03.031

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Clinical and basic research evidence suggests a critical role of local hemodynamic microenvironments in atherogenesis. Disturbed flow with a low and reciprocating shear stress in arterial branches and curvatures induces endothelial dysregulation and damage, leading to atherosclerosis. In contrast, laminar flow with a high and unidirectional shear stress in the straight parts of the arteries elicits an atheroprotective effect on the vasculature. Effective treatment against endothelial dysregulation and the consequent atherogenesis requires the identification of new shear-responsive molecules and the development of drugs targeting these molecules. In this review, we focus on the role of shear-responsive transcription factors and cofactors in endothelial dysfunction and atherosclerosis, and highlight future direction for prevention and treatment of atherosclerosis by targeting these factors.

Effect of miR-216a on macrophages derived from human peripheral blood mononuclear cells

HAN Shuang, CHEN Yu, ZHANG Wei-li

2022, 42(3):  384-388.  doi:10.16352/j.issn.1001-6325.2022.03.034

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Objective To study the role of miR-216a in regulating the characteristics of macrophages derived from peripheral blood mononuclear cells (PBMCs). Methods Human PBMCs were isolated and cultured. The polarization model of M1 and M2 macrophages were established in vitro, and flow cytometry and real-time quantitative PCR were used to detect the cell surface markers of macrophage. Furthermore, miR-216a mimics were transfected into M2 macrophages, and the surface markers of M2 macrophage and inflammatory factors were examined. Results After overexpression of miR-216a, the expression of surface marker CD163 of M2 macrophage was significantly increased, the expressions of inflammatory cytokines TNFα and IL-1β were decreased, and the expressions of anti-inflammatory cytokines TGF-β and IL-10 were increased(P＜0.05). Conclusions miR-216a could promote the polarization of human PBMCs-derived macrophages into M2 phenotype and inhibit the expressions of pro-inflammatory cytokines.

Original Articles

Role of mitochondrial permeability transition pore in myocardial ischemia-reperfusion injury in rat models with zinc deficiency

LIAN Ting, ZHANG Rui-jun, XIONG Xiao-lan, ZHANG Xiao-ya, YU Tao, ZHANG Shi-zhong

2022, 42(3):  389-394.  doi:10.16352/j.issn.1001-6325.2022.03.030

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Objective To investigate whether zinc deficiency may induce myocardial ischemia-reperfusion (I/R) injury through mitochondrial permeability transition pore (mPTP). Methods The zinc-deficient rat model was established by feeding with a zinc-deficient fodder, myocardial I/R injury was induced with isolated rat hearts which were subjected to 30 min of global ischemia followed by 120 min of reperfusion. Left ventricular catheterization was used to detect cardiac function. The lactate dehydrogenase (LDH) was spectrophotometically measured; and 2,3,5-triphenyl-tetrazolium(TTC) staining method was applied to measure the area of myocardial infarction; Mitochondrial swelling induced by calcium was adapted as a method to examine the opening degree of mPTP. Results Rats with zinc deficiency showed an increased myocardial infarct size and LDH release (P＜0.05), which was associated with a decreased recovery rate of left ventricular contractility(P＜0.05). Compared to control rats, mPTP opening reduced in mitochondria of the heart of zinc-deficient rats before ischemia(P＜0.05), but increased significantly after 30 min ischemia (P＜0.05). The myocardial I/R injury in both control and zinc-deficient rats was significantly attenuated by inhibition of mPTP opening(P＜0.01). Conclusions Zinc deficiency may increase the myocardial I/R injury in rats through increase of mPTP opening.

Knockout of EZH2 inhibits migration and invasion of human cervical cancer cell lines

CHEN Qian, LIU Xian, CUI Nan, ZHAI Yang

2022, 42(3):  395-400.  doi:10.16352/j.issn.1001-6325.2022.03.004

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Objective To explore the effect of EZH2 on the migration and invasion of human cervical cancer cell lines and its mechanism. Methods CRISPR/Cas9 was used to knockout EZH2 expression in HeLa and SiHa cell lines. The migration was detected by scratch test, and the migration and invasion in vitro was detected by Transwell migration and invasion test. The STAT3 mRNA level was detected by real-time fluorescence quantitative PCR, and the expression of STAT3 and p-STAT3 proteins was detected by Western blot. Results CRISPR/Cas9 could knockout EZH2 expression. The wound healing rate of HeLa and SiHa cells in EZH2 knockout group at 24 h and 48 h, and the counting of transmembrane cell migration and invasion in Transwell chamber were lower than those in control group(P＜0.05). Compared with the control group, the protein level of p-STAT3 in EZH2 knockout group was significantly decreased (P＜0.05). Conclusions EZH2 plays an important role in promoting the migration and invasion of cervical cancer cells, which may be related to the targeted regulation of STAT3 pathway, and is a potential target in the treatment of cervical cancer.

Increased serum IL-1 level and activated JAK2/STAT3 signaling pathway of cancer tissue in patients with gastric cancer

NIU Shi-wei, SU Yun, GONG Hong-xia, LI Ting-ting, ZENG Yuan-ding, ZHANG Han, NIE Peng

2022, 42(3):  401-405.  doi:10.16352/j.issn.1001-6325.2022.03.035

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Objective To explore the role of elevated serum interleukin 1(IL-1) levels in patients with gastric cancer and activation of JAK2/STAT3 signaling pathway in tissues of gastric cancer. Methods A total of 30 gastric cancer surgery patients and 30 healthy subjects were selected from Wuwei Cancer Hospital of Gansu Province; The levels of IL-1 and IL-10 in serum was determined by enzyme-linked immunosorbent assay (ELISA); Immunohistochemistry and Western blot were used to detect the contents of Bcl-2, Bax and JAK2,STAT3, pJAK2, pSTAT3, Bcl-2, Bax in tissues. The contents of JAK2, STAT3, Bcl-2 and Bax genes were measured by RT-qPCR. Results Compared with the control group, the level of IL-1 in the serum of patients with gastric cancer was up-regulated, and the level of IL-10 was down-regulated(P＜0.05); The content of Bcl-2 in cancer tissue was significantly higher than that in the normal group and the adjacent group, but the Bax content showed an opposite profile(P＜0.05). Compared with the control group, JAK2, STAT3, pJAK2, pSTAT3 and Bcl-2 protein content in cancer tissue and adjacent tissues were increased, while the content of Bax was decreased(P＜0.05). Compared with the adjacent tissues, the JAK2, STAT3, pJAK2, pSTAT3 and Bcl-2 protein contents in cancer tissues increased, while the Bax content was decreased(P＜0.05). Compared with the control group, the contents of JAK2, STAT3 and Bcl-2 mRNA in cancer tissues and adjacent tissues were increased, while Bax mRNA was decreased(P＜0.05). Compared with adjacent tissues, the contents of JAK2, STAT3 and Bcl-2 mRNA in cancer tissues increased, while the content of Bax mRNA was decreased(P＜0.05). Conclusions Abnormal elevation of proinflammatory factors and abnormal activation of JAK2/STAT3 signaling pathway are closely related to the occurrence and development of gastric cancer.

Establishment of cell models for Blau syndrome

SONG Hao-xin, YE Cai-ying, ZHU Lei

2022, 42(3):  406-410.  doi:10.16352/j.issn.1001-6325.2022.03.036

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Objective To establish a stable and reliable cell models of Blau syndrome (BS)in vitro. Methods RAW264.7, iBMDM and THP-1 cells were used as the research objects and then stimulated by muramyl dipeptide (MDP) or L18-MDP. Meanwhile, positive drugs etanercept (ETN) and GSK583 treatment groups were set to investigate the response of the model to evaluate effectiveness. Cell culture supernatant was collected after 22 h, and the level of tumor necrosis factor-α (TNF-α) was determined by enzyme linked immunosorbent assay (ELISA). Results The secretion of TNF-α from RAW264.7 and iBMDM cells was significantly increased after stimulation by 10 μg/mL MDP(P＜0.01), while ETN and GSK583 inhibited the increase of TNF-α induced by MDP(P＜0.01). After THP-1 cells were induced into THP-1-Mφ by PMA, 0.2 and 1 μg/mL L18-MDP increased the level of TNF-α in the culture supernatant(P＜0.01). When ETN was added along with 0.2 μg/mL L18-MDP, the secretion of TNF-α was significantly inhibited (P＜0.01). Conclusions RAW264.7,iBMDM and THP-1 cells may develop good cell models for BS in vitro. The model has the advantages of simplicity, convenience, easy repetition and strong controllability, which lay a foundation for further research on the pathogenesis of BS and screening and evaluation of therapeutic drugs.

Sulforaphane alleviates ischemia- reperfusion injury of heart transplantation in rat models

WANG Shuai, WANG Jian-jun, SUN Xiu-hong, WU Peng-yu, CUI Zhi-cheng, LI Zhan-qing

2022, 42(3):  411-416.  doi:10.16352/j.issn.1001-6325.2022.03.029

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Objective To investigate the effect of sulforaphane on ischemia-reperfusion injury(I/RI) and the role of PI3K/Akt/GSK-3β pathway in heart transplantation of rats. Methods The rat model of allograft heart transplantation was established as the control group.The survival rate,the level of serum myocardial enzymes,apoptosis of myocardial cells and the swelling degree of myocardial mitochondria were compared with sulforaphane injection group (2.5 mg/kg).Western blot was used to detect the expression of apoptotic molecules caspase-3,Bax and PI3K/Akt/GSK-3β. Results The survival rate of sulforaphane group was significantly higher than that of control group(P＜0.05).The serum concentrations of LDH,CK,CK-MB and TNT in sulforaphane group were lower than those in control group with a time dependent manner at different treatment time (6,12,24 and 48 h).The expressions of caspase-3 and Bax were significantly decreased (P＜0.01),and the degree of mitochondrial swelling was also improved compared with the control group. Conclusions Sulforaphane may inhibit myocardial apoptosis by activating PI3K/Akt/GSK-3β pathway and improve I/RI after heart transplantation as shown by animal model.

Ketorolac tromethamine inhibits inflammation in rat models with knee osteoarthritis

LI Rong, ZHU Han-yue

2022, 42(3):  417-422.  doi:10.16352/j.issn.1001-6325.2022.03.028

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Objective To investigate the effect of ketorolac tromethamine (KT) on the inflammatory pain caused by knee osteoarthritis (KOA) of rat model and to explore potential mechanism related to Toll-like receptor 4 (TLR4)/nuclear factor-κB (NF-κB) inflammatory pathway. Methods The rats were divided into: control group, model group, low (1 mg/kg),high (4 mg/kg) dose KT group,TAK-242 group (TLR4 antagonist,1 mg/kg) by random number table method; Except for the control group, the other groups used the modified Hulth method to replicate the KOA model of the right knee in rats; KT was administered by intramuscular injection once a day, and TAK-242 was administered by tail vein injection twice a week. The swelling and activity of the knee joint of the rats were recorded, and the spontaneous pain behavior gait and the thermal pain threshold were measured; the synovial tissue was sampled, and the histopathological changes and fibrous tissue proliferation were detected by microscopy with HE staining and Masson staining; The expression of interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α),neuritin expression and the TLR4,NF-κB,phosphorylated NF-κB (p-NF-κB),osteopontin (OPN),integrin metalloproteinase 4 (ADAM4) proteins expression in synovium tissue were detected. Results Compared with the control group, the model group rats had more severe pathological symptoms of KOA, such as joint swelling, pain, synovial inflammatory damage and fibrous tissue hyperplasia, the IL-1β and TNF-α levels and the neuritin expression,the TLR4/NF-κB p65 pathway and related proteins OPN,ADAM4 expression in synovium increased (P＜0.05).Compared with model group, the pathological symptoms such as KOA pain and synovial inflammation of rats in low, middle and high dose KT groups were alleviated, the TLR4/NF-κB p65 pathway and related proteins expression in synovium decreased (P＜0.05). Conclusions KT can relieve the synovial inflammation and pain symptoms of KOA rats; Its relieving effect may be related to blocking the activation of TLR4/NF-κB pathway.

miR-188 regulates TGF-α to inhibit proliferation and migration of osteosarcoma cells

LIU Wei, LIU Xiao-feng, ZHAO Bao-hui

2022, 42(3):  423-428.  doi:10.16352/j.issn.1001-6325.2022.03.003

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Objective To explore the effects of miR-188 and transforming growth factor α (TGF-α) on the proliferation and migration of osteosarcoma cells. Methods The expressions of miR-188 and TGF-α in human osteosarcoma tissues and adjacent tissues were detected. TGFA was detected as the target gene of miR-188 by TargetScan and dual luciferase reporter gene assay. The migration ability of osteosarcoma cells line MG63 was examined by cell scratch assay. The xenograft model of osteosarcoma was established in nude mice and the effect of miR-188 was observed in a xenograft model of osteosarcoma of nude mice. Results The expression of miR-188 was decreased in osteosarcoma tissues (P＜0.01). miR-188 specifically bound to the 3′-UTR of TGFA and negatively regulated its expression. The transfection of miR-188 mimic could inhibit the migration ability of MG63 cells,while the inhibitor could promoted the migration ability,but the up-regulation of TGF-α and the expression of interfered TGF-α could reduce the inhibition effect. In a nude mouse model of osteosarcoma xenograft,over-expression of miR-188 reduced the inhibitory effect (P＜0.01), while down-regulating the expression of miR-188 expression had the opposite effect. TGF-α expression was significantly increased in osteosarcoma tissues (P＜0.01),while silence protiferation of osteosarcoing was decreased. Conclusions miR-188 may inhibit the proliferation and migration of MG63 cells by targeting the expression of TGF-α.

Gelsemine inhibits proliferation and invasion of rheumatoid arthritis synovial fibroblast through down-regulating circ_001680

LIU Xin-bing, ZHANG Mei-hong

2022, 42(3):  429-435.  doi:10.16352/j.issn.1001-6325.2022.03.027

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Objective To investigate the effect of gelsemine on the proliferation, migration and invasion of rheumatoid arthritis endothelial cells (RASFs) and its possible mechanism. Methods RASFs were isolated and cultured. After RASFs were intervened with 50, 100, 200 μg/mL gelsemine for 24 h or small interfering RNA or over-expression vector of circular RNA circ_001680 was transfected into RASFs, cell proliferation was measured by CCK-8 method and clone formation test, cell migration was detected by scratch test, cell invasion was examined by Transwell, the protein expression of E-cadherin and N-cadherin was detected by Western blot. The protein expression of circ_001680 was examined by RT-qPCR. Results Gelsemine or interference with circ_001680 expression reduced the A value, the number of clones, the healing rate of scratches and the number of invaded cells of RASFs and the protein expression of N-cadherin in cells (P＜0.05), and promoted the protein expression of E-cadherin (P＜0.05). Gelsemine reduced the expression of circ_001680 in RASFs (P＜0.05), and the over-expression of circ_001680 reduced the inhibitory effect of gelsemine on the proliferation, migration and invasion of RASFs. Conclusions Gelsemine may inhibit the proliferation, migration and invasion of RASFs, and its mechanism is potentially related to the down-regulation of circ_001680 expression.

Cinobufagin inhibits proliferation of human nasopharyngeal carcinoma cell lines CNE2 and HONE1

ZHAO Xia, CAI Qing-chun, DING Rui-min, ZHANG Qian

2022, 42(3):  436-440.  doi:10.16352/j.issn.1001-6325.2022.03.026

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Objective To evaluate the biological effect of cinobufagin(Cin) on nasopharyngeal carcinoma (NPC) cells lines(CNE2 and HONE1) and the mechanism of its efficacy. Methods CNE2 and HONE1 cells were treated with different concentrations (0, 5, 15 and 45 mg/L) of Cin for 24, 48 and 72 h respectively and cell viability was detected by CCK-8 assay. After 48 hrs, cell morphology was observed by phase contrast microscopy; cell apoptosis was detected by TUNEL fluorescence staining; the level of reactive oxygen species (ROS) in cells were assessed by DCF fluorescence assay; the expression levels of apoptosis-related proteins were detected by Western blot. Results Cin inhibited the cell activity of NPC cells in a concentration-dependent manner (P＜0.05, P＜0.01 or P＜0.001) and the cells showed filament and sparse morphology. Cin promoted apoptosis and increased intracellular ROS level in NPC cells (P＜0.05, P＜0.01 or P＜0.001). Cin inhibited the expression of Bax and mitochondrial cytochrome c (Cytc) (P＜0.05, P＜0.01 or P＜0.001) but promoted the expression of Bcl-2, cleaved caspase-3 and cytoplasmic Cytc (P＜0.01 or P＜0.001). Conclusions Cin can inhibit cell proliferation and promote apoptosis of NPC cells.

LRCH1 maintains the function of regulatory T cells in mouse models of lupus nephritis

QIN Zhi-hui, TAN De-min, WANG Wang-zhen, ZHANG Yao

2022, 42(3):  441-447.  doi:10.16352/j.issn.1001-6325.2022.03.006

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Objective To evaluate the expression and effect of leucine rich repeats and calponin homology domain containing 1 (LRCH1) in regulatory T cells (Tregs) from lupus nephritis. Methods The 6-month old C57BL/6 and MRL/lpr mice were used to enrich CD45⁺CD4⁺CD25⁺CD127^low Tregs from the spleens and kidneys by flow cytometry. RT-qPCR was applied to measure LRCH1 mRNA in these cells. Exogenous Tregs were adoptively transferred into C57BL/6 mice and MRL/lpr mice, followed by detection of mRNA of LRCH1,IL-10. TGFβ(transforming growth factor beta) in exogenous Tregs in the spleen and kidney of recipient mice using RT-qPCR. Furthermore, normal Tregs were transfected with siRNAs to silence Lrch1 expression and followed by the evaluation of Treg-induced suppression of conventional T cell proliferation with flow cytometry. Immunoblotting was performed to find the activation of IL-2 signaling. Results Compared with wild type mice, Tregs up-regulated LRCH1 by 4.13 folds in the kidney of MRL/lpr mouse models. The adoptive transfer assay indicated that the lupus nephritis microenvironment induced 1.98-fold increase of LRCH1, 13.83-fold increase of IL-10, and 3.58-fold increase of transforming factor beta in Tregs. Lrch1 knockdown reduced IL-2-induced p-STAT5 by half, thus inhibiting IL-2 signaling, down-regulating Foxp3 expression, and weakening Tregs-induced suppression of conventional T cell proliferation. Conclusions LRCH1 is a positive regulator of immunosuppressive activity of infiltrating Tregs in mouse models of lupus nephritis.

Ropivacaine inhibits proliferation, migration and invasion of human liver cancer cell line Hep3B

SUN Quan-peng, FAN Chao, ZHAN De-xi, FAN Yi-ming, SUN Wei-feng

2022, 42(3):  448-453.  doi:10.16352/j.issn.1001-6325.2022.03.025

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Objective To study the effects of ropivacaine on the biological behaviour of liver cancer cell line Hep3B and to explore its molecular mechanism. Methods Hep3B cells were divided into the control group, ropivacaine group, si-NC group, si-LINC00853 group, pcDNA-NC group, pcDNA-LINC00853 group,(ropivacaine+pcDNA-NC) group and (ropivacaine +pcDNA-LINC00853) group. Cell Counting Kit(CCK-8), Transwell assays, and RT-qPCR were applied to detect cell proliferation, migration, invasion and LINC00853 expression. Western blot was used to analyze the expression of proliferating cell nuclear antigen (PCNA), matrix metalloproteinase 2(MMP2) and MMP9 proteins. Results Compared with the NC group, the proliferation inhibition rate of Hep3B cells in the ropivacaine group was elevated, cell migration and invasion were inhibited. The expression of PCNA, MMP2, MMP9 and LINC00853 was reduced (P＜0.05). Compared with the si-NC group, the proliferation inhibition rate of Hep3B cells in the si-LINC00853 group was elevated, cell migration and invasion were reduced and the expression of PCNA, MMP2, MMP9 protein was decreased (P＜0.05). Compared with the pcDNA-NC group, the proliferation inhibition of Hep3B cells in the pcDNA-LINC00853 group was reduced, cell migration and invasion were increased. The expression of PCNA, MMP2, and MMP9 protein was increased (P＜0.05). Compared with the (ropivacaine+ pcDNA-NC) group, the proliferation inhibitio of Hep3B cells in the (ropivacaine+ pcDNA-LINC00853) group was reduced, cell migration and invasion were increased and the expression of PCNA, MMP2 and MMP9 protein increased (P＜0.05). Conclusions Ropivacaine can inhibit liver cancer cell line Hep3B proliferation, migration and invasion. Its mechanism may be related to the down-regulation of LINC00853 expression.

LncRNA UNC5B-AS1 targeting miR-339-5p regulates the proliferation and apoptosis of human lung adenocarcinoma cell line A549

ZHANG Jian, ZHANG Ji-jiong

2022, 42(3):  454-460.  doi:10.16352/j.issn.1001-6325.2022.03.022

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Objective To study the effect of lncRNA UNC5B-AS1 targeting miR-339-5p on the proliferation and apoptosis of lung cancer cells and its molecular mechanism. Methods The human lung adenocarcinoma cell line A549 was cultured in vitro, and si-NC, si-UNC5B-AS1, miR-NC, miR-339-5p mimics, si-UNC5B-AS1 and anti-miR-NC, si-UNC5B-AS1 and anti-miR-339-5p were transfected into A549 cells respectively. Real-time fluorescent quantitative PCR (RT-qPCR) was used to detect the expression of UNC5B-AS1 and miR-339-5p in lung cancer tissues and in adjacent tissues; Cell counting kit (CCK-8) was used to detecte cell proliferation; The targeted regulation relationship between UNC5B-AS1 and miR-339-5p was examined by dual luciferase reporter gene experiment; Western blot was used to detect the expression of cyclin D1 (cyclin D1), p21 protein lymphoma-2(Bcl-2), and B lymphoma-2 related protein (Bax). Results Compared with adjacent tissues, the expression level of UNC5B-AS1 in lung cancer tissue was significantly increased (P＜0.05), and the expression level of miR-339-5p was significantly decreased(P＜0.05). Compared with the si-NC group, the cell viability of the si-UNC5B-AS1 group was significantly reduced (P＜0.05), the apoptosis rate was significantly higher(P＜0.05). Compared with the miR-NC group, the cell viability of the miR-339-5p group was significantly reduced (P＜0.05), and the apoptosis rate was significantly increased(P＜0.05). Compared with the (si-UNC5B-AS1+anti-miR-NC)group, the cell viability of the (si-UNC5B-AS1+anti-miR-339-5p) group was significantly increased (P＜0.05); the apoptosis rate was significantly reduced (P＜0.05). Conclusions Interference with UNC5B-AS1 promotes apoptosis and inhibits cell proliferation of human lung adenocarcinoma cancer cell line A549 by up-regulating the expression of miR-339-5p.

Effect of SLC4A10 expression on prognosis of patient with hepatocellular carcinoma

HUANG Rong, ZHANG Jian, CHE Xu

2022, 42(3):  461-466.  doi:10.16352/j.issn.1001-6325.2022.03.024

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Objective To analyze the effect of SLC4A10 expression on the prognosis of patients with hepatocellular carcinoma(HCC)by bioinformatics, and to confirm its expression in HCC tissue and normal tissue. Methods The expression of SLC4A10 in HCC and in normal tissues was analyzed by TCGA database, and verified by real-time quantitative PCR. Kaplan-Meier(K-M) curve was applied to assess survival profile . Univariate and multivariate Cox regression analysis was applied to address the influence of independent factors on overall survival(OS)and disease free survival (DFS). PPI database was used to construct protein interaction network and function enrichment analysis was used to explore the regulatory mechanism of SLC4A10 in HCC. Results Significantly decreased transcriptional SLC4A10 expression was found in HCC samples. Decreased SLC4A10 mRNA expression was significantly associated with advanced pathological parameters and with shorter OS and DFS in TCGA cohorts. Functional annotations indicated that SLC4A10 was involved in the most significant hallmarks including transepithelial transport, cell volume homeostasis and sodium ion import across plasma membrane. Conclusions The decreased SLC4A10 expression is significantly correlated with aggressive progression and poor survival of HCC patients. These data suggest that SLC4A10 may act as a promising prognostic marker or therapeutic target in HCC.

Effect of pitavastatin on levels and functions of HDL-C in patients with stable coronary artery disease and dyslipidemia

WANG Hui-juan, WANG Ji-hong, ZHAO Xing-shan, LIU Wei

2022, 42(3):  467-471.  doi:10.16352/j.issn.1001-6325.2022.03.023

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Objective To evaluate the effect of pitavastatin on high-density lipoprotein cholesterol (HDL-C) level and cholesterol efflux and antioxidative properties of HDL in patients with dyslipidemia and stable coronary disease. Methods The study population included 60 patients with stable coronary disease and dyslipidemia. They were randomly allocated to treatment by pitavastatin 2-4 mg per day or atorvastatin 10-20 mg per day and followed up for six months. The cholesterol efflux and activities of antioxidative enzymes paraoxonase-1 (PON-1) were evaluated before and after statins treatment. Results The effect of the 6-month treatment with pitavastatin on serum HDL-C level was significantly greater than that of atorvastatin (% change: pitavastatin 9.0% vs atorvastatin 1.8%, P＜0.01). HDL induced cholesterol efflux increased significantly after the 6- month treatment with pitavastatin while it not was affected by atorvastatin treatment. However, HDL-associated PON-1 in both pitavastatin group and atovastatin group all increased. The activity of PON-1 increased by 21% with pitavastatin and 24% with atorvastatin, while the activity of arylesterase increased by 18% with pitavastatin and 17% with atorvastatin (P＜0.001). Conclusions Pitavastatin elevates the HDL-C level, enhances the cholesterol efflux and antioxidative effect of HDL. Therefore, pitavastatin increases the amount of functional HDL without attenuating HDL quality.

Circ-MALAT1 targeting miR-101 inhibits invasion and migration of bladder cancer cell line SW780

YANG Mang-zhuang, YANG Xu-dong, WANG Xiao-long

2022, 42(3):  472-478.  doi:10.16352/j.issn.1001-6325.2022.03.020

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Objective To explore the function of circ-MALAT1 targeting miR-101 in the inhibition of the invasion and migration of bladder cancer cells. Methods Bladder cancer cell line SW780 was divided into si-NC group (transfection si-NC), si-circ-MALAT1 group (transfection si-circ-MALAT1), miR-NC group (transfection miR-NC), miR-101 group (transfection miR-101 mimics), si-circ-MALAT1+anti-miR-NC group(co-transfection si-circ-MALAT1 and anti-miR-NC)and si-circ-MALAT1+anti-miR-101 group(co-transfection si-circ-MALAT1 and anti-miR-101). RT-qPCR was used to examine the expression of circ-MALAT1 and miR-101; Transwell method was used to detect cell invasion and migration; The matrix metalloproteinase 2 (MMP-2) and matrix metalloproteinase 9 (MMP-9),phosphorylated phosphatidylinositol 3-kinase (p-PI3K), phosphorylated protein kinase B(p-AKT) protein expression were measured by Western blot; Dual luciferase reporter gene experiment was applied to detect the targeting relationship between circ-MALAT1 and miR-101. Results Compared with adjacent tissues, the expression of circ-MALAT1 in bladder cancer tissue was significantly increased (P＜0.05)and the expression of miR-101 was significantly decreased (P＜0.05). Compared with the si-NC group, the expression of circ-MALAT1 in the si-circ-MALAT1 group was significantly reduced(P＜0.05)and the counting number of invasion and migrating cells was significantly reduced (P＜0.05), the expression of p-PI3K and p-AKT protein was significantly reduced(P＜0.05). Compared with the miR-NC group, the expression of miR-101 in the miR-101 group was significantly increased(P＜0.05)and the number of invasion as well as migrating cells were significantly reduced(P＜0.05).Compared with the (si-circ-MALAT1+anti-miR-NC)group, the(si-circ-MALAT1+anti-miR-101)group showed more cell invasion and migration (P＜0.05), the expression of p-PI3K and p-AKT protein was significantly increased(P＜0.05). Conclusions Reduction of circ-MALAT1 expression and up-regulation miR-101 inhibit the invasion and migration of bladder cancer cells line SW780. The mechanism may be related to the PI3K/AKT signaling pathway.

Gonadotropin releasing hormone antagonist protocol brings about more available embryos than agonist long protocol

ZHAO Shan-ke, FU Yao, CHU Da-peng, FU Lei, HAN Shuo, BAI Xue-yan, LI Yuan, ZHOU Wen-hui

2022, 42(3):  479-483.  doi:10.16352/j.issn.1001-6325.2022.03.005

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Objective To compare the effect on embryo quality from gonadotropin releasing hormone(GnRH) antagonist protocol and from GnRH agonist long protocol, so as to find which protocol works better on embryo quality. Methods A retrospective analysis was performed with a total of 1 095 long protocol cycles and 1 511 antagonist protocol cycles from January 2016 to December 2019. The corresponding fertilization, embryo quality, pregnancy outcomes of the two groups, agonist vs antagonist, were calculated and analyzed. Results Antagonist group displayed significantly higher available embryo rate (29.94% vs 31.59%), good-quality two-pronuclear(2PN) embryo rate (44.67% vs 46.86%), available blastocyst/OPU(ovum pick-up)(1±2 vs 1±2) and available blastocyst rate (25.62% vs 29.66%) than the long protocol group. Conclusions Antagonist protocol may bring about more available embryos than agonist protocol. Together with a similar pregnancy outcome with agonist protocol and its efficacy in reducing ovarian hyperstimulation syndrome(OHSS) risk, antagonist protocol is the first choice for clinical contralled ovarian stimulation(COS) protocol.

Clinical Sciences

Clinical pregnancy outcome analysis of 1 169 frozen-thawed embryo transfer cycles

LI Tian-feng, SHUAI Zhen-hong, ZHANG Wan-jing, LI Xue-mei

2022, 42(3):  484-487.  doi:10.16352/j.issn.1001-6325.2022.03.021

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Objective To investigate the factors in frozen-thawed embryo transfer (FET) cycles that affecting the pregnancy outcome. Methods The clinical data of 1 169 couples who underwent FET in Shenzhen Maternity and Child Healthcare Hospital from January, 2017 to December, 2019 were retrospectively analyzed. Univariate analysis and Logistic multivariate regression analysis were performed to identify the factors which influence pregnancy outcome of FET cycles. Results The clinical pregnancy rate of 1 169 FET cycles was 50.13%. Univariate analysis showed that female patients ＞35 years old or infertility duration ＞ 6 years significantly had a reduced embryo implantation rate and clinical pregnancy rate (P＜0.01). But the embryo implantation rate and clinical pregnancy rate were increased by blastocyst or multiple embryos transfer (P＜0.01). Multiple embryo transplantation increased embryo implantation rate (P＜0.05) and endometrial thickness ＞12 mm increased clinical pregnancy rate (P＜0.05). Logistic multivariat regression analysis showed that elder age and decreasing number of optimal embryo transplantation were risk factors for clinical pregnancy rate (P＜0.01). Conclusions Patients age, embryo age and the number of transplanted embryos of high quality are the main factors affecting the pregnancy outcome of FET. Elder age is a risk factor for clinical pregnancy outcome.

Technology and Methodology

Preparation and mixed-samples detection validation of nucleic acid detection kit for SARS-CoV-2

ZHAO Qian-qian, LIU Shuang-shuang, ZHOU Yun-ying, SHI Gui-zhi, WANG Yun-shan

2022, 42(3):  488-493.  doi:10.16352/j.issn.1001-6325.2022.03.018

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Objective A novel corona virus(SARS-CoV-2) nucleic acid detection kit was developed and performance had been optimized for large-scale population screening in China. The effect of mixed detection of multiple samples was analyzed aiming at providing theoretical guidance for clinical use. Methods The primers and probe sequences for ORF1ab and N announced by China Center for Disease Control and prevention and the primers and probe sequences for RNase P as control gene were synthesized. Single variable optimization and orthogonal experiment were used to get the best concentration ratio of primers and probes for ORF1ab, N and RNase P in fluorescent RT-qPCR detection system. The accuracy, specificity and sensitivity of the kit were investigated. The positive samples and negative samples were collected and mixed according to different proportions, and the SARS-CoV-2 was detected by this kit and the commercial reference kit respectively. The detection concentration for different targets and the amplification Ct value were compared. Results The positive coincidence rate, negative coincidence rate and specificity of the kit were 100%, respectively。And the sensitivity was 1×10³ copies/mL. The detection sensitivity of this kit for ORF1ab target gene was 1/20, higher than that of reference kit (1/10), and the amplification Ct value with same template concentration was lower than that of reference kit (P＜0.05). Conclusions This kit has great advantages in screening multi samples detection of new corona virus nucleic acids because of high sensitivity to ORF1ab target gene, and suitable for in large-scale population.

Mini Reviews

Research progress on estrogen and its receptors in occurrence and advance of Crohn's disease

WU Guo-zhi, AN Li-na, LIU Min, WANG Yu-ping, GUO Qing-hong

2022, 42(3):  494-497.  doi:10.16352/j.issn.1001-6325.2022.03.017

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Crohn's disease (CD) is a subtype of inflammatory bowel diseases and may be resulted from endocrine disorders. Estrogen and its receptors exert an important role in the pathogenesis of CD. Studies have revealed that estrogen can down-regulate the expression of inflammatory mediators while estrogen supplementation helps reduce the severity of inflammatory response. The expression of receptors (GPER, ERα and ERβ) also influence local immune responses, and the imbalance of them mediate intestinal inflammation. Nowadays, estrogen and its receptors are expected to become a new rarget in the research of CD.

Research progress of metallothionein in metabolic diseases

YANG Yan

2022, 42(3):  498-501.  doi:10.16352/j.issn.1001-6325.2022.03.019

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Metallothionein (MT) is a kind of low molecular metal binding proteins rich in cysteine. It plays a protective role in obesity, non-alcoholic fatty liver disease (NAFLD), diabetes mellitus(DM) and its complications. It has many functions such as anti-oxidative stress, anti-inflammatory, and inhibiting cell apoptosis.

Research progress on the role of myocardial cell calcium homeostasis dysregulation in heart failure

LIU Feng-yu, HAN Wei

2022, 42(3):  502-506.  doi:10.16352/j.issn.1001-6325.2022.03.008

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Calcium homeostasis of cardiomyocytes depends on the regulation of calcium(Ca²⁺) transportation through plasma membrane, sarco/endoplasmic reticulum(SR) and mitochondria. One of the main characteristics of heart failure is calcium homeostasis, including cytosolic calcium overload and SR calcium leakage, which leads to transient decrease of myocardial calcium and decrease of contractility.

Advances on the roles of circular RNA in breast cancer

HE Xiao, HU Wei-jie, LYU Wen-chang, REN Yu-ping, WU Min, WU Yi-ping, ZHANG Qi

2022, 42(3):  507-511.  doi:10.16352/j.issn.1001-6325.2022.03.007

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Circular RNA (circRNA) is a kind of non-coding RNA with circular structure and is found in a variety of cell types. CircRNA is characterized by diversity, specificity, conservatism and stability. As an emerging non-coding regulatory molecules, circRNA can regulate the proliferation and metastasis of breast cancer (BC) mainly through the mechanism of competitive endogenous RNA, and is involved in a variety of chemotherapy resistance and affect the efficacy. Elucidating the functions and mechanisms of circRNA in BC may provide novel and effective strategies for the diagnosis and treatment of BC.

Research advances in the roles of local anesthetics in tumor metastasis and recurrence

LIU Chang, GUO Xiang-yang

2022, 42(3):  512-515.  doi:10.16352/j.issn.1001-6325.2022.03.015

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Anesthetics are commonly used in the perioperative treatment of tumor. The choice of different anesthetics and anesthesia techniques will affect the proliferation, metastasis, recurrence and prognosis of cancer. Perioperative applications of local anesthetics can not only reduce the amount of opioids, but also inhibit tumor metastasis and recurrence by blocking sodium channels of cancer cells, changing epigenetics, decreasing inflammation and improving immune function. The exploration of the mechanism of local anesthetics in tumor metastasis and recurrence provides the new strategies for the optimal use of local anesthetics in cancer patient's perioperatively chemotherapy.

Research progress of gut microbiota in the pathogenesis of gestational diabetes mellitus

HU Xian-liang, WAN Yan, FENG Ji

2022, 42(3):  516-519.  doi:10.16352/j.issn.1001-6325.2022.03.016

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Gut microbiota has an important influence in the pathogenesis of gestational diabetes mellitus(GDM).Gut microbiota rises insulin resistance,hypoinsulinemia,the disorder of blood glucose resulted from damage of the gut membrane barriers and from releasing metabolite,adipokine and inflammatory factors and then interfere the gut immune functions, finally result in GDM. The probiotic intake can effectively improve the disorders of gut microbiota and blood glucose of GDM patients, furthermore improve the pregnancy outcomes.

Progress in the study of the epigenetic regulation of cardiac hypertrophy

ZHENG Jing-jing, BU Ning, ZHAO Zhao, ZHANG Jia-jing, YU Hui, ZHAO Yu-hang

2022, 42(3):  520-524.  doi:10.16352/j.issn.1001-6325.2022.03.011

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Myocardial hypertrophy is an independent risk factor for various serious cardiovascular diseases.Epigenetic mechanisms such as DNA methylation, histone modification, and RNA methylation are involved in the pathogenesis and development of cardiac hypertrophy.To understanding the role of epigenetic regulation on cardiac hypertrophy contributes to to development of an effective protocol and provide scientific basis for the prevention and early treatment of cardiac hypertrophy.

Medical Education

Construction and practice of basic medical comprehensive PBL curriculum system under the background of “first-class curriculum”

LI Juan, LI Min-hui, LIN You-sheng, HU Xiao-song, YANG Ping

2022, 42(3):  525-528.  doi:10.16352/j.issn.1001-6325.2022.03.013

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As one of the first batch of universities selected as members of national “Outstanding Physician Education and Training Program”,Chengdu Medical College has initiated integrated and organ-based remodeling curriculum since 2016, and set up “Comprehensive Basic Medicine problem-based learning(PBL) Course” for the pilot class of outstanding doctors. Through PBL learning, students are guided to comprehensively apply the basic medical knowledge that they have learned to explain clinical phenomena and mechanisms, connecting the preceding and the following, so as to lay a solid foundation for subsequent clinical learning and realize the continuity and systemization of medical knowledge. After several years of practice. Some experience and achieved good teaching effect have been accumulated.

Effectiveness of clinical classroom teaching evaluation system

ZHAO Jun, XU Yuan, PAN Hui, LUO Lin-zhi

2022, 42(3):  529-532.  doi:10.16352/j.issn.1001-6325.2022.03.014

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In order to systematically supervise the quality of clinical classroom teaching of eight-year program of clinical medicine at Peking Union Medical College, a classroom teaching evaluation system was established. The effectiveness of the evaluation system was tested from three aspects including the effectiveness of the content of the evaluation system, the internal and external influence factors on the evaluation and the application of the evaluation results. A questionnaire was designed to carry out a random survey among the students. The current classroom teaching evaluation system is basically effective though there are still some weaknesses need to be improved.

Implementation of designing clinical research training in Tibet Autonomous Region People's Hospital

WANG Feng-dan, YANG Xiao, PANG Hai-yu, ZHU Ye-yi, PARK Meyeon, YANG Dun-gan

2022, 42(3):  533-536.  doi:10.16352/j.issn.1001-6325.2022.03.009

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Objective To evaluate the research training and experience of medical staff in Tibet Autonomous Region People's Hospital (TARPH), and their feedback after attending a series course of designing clinical research (DCR). Methods After surveying about the research training and experience of medical staff in TARPH, two doctors from Peking Union Medical College Hospital(PUMCH) who were sent to aid TARPH taught DCR course for 12 weeks. After completion of the course, participants joined a questionnaire survey to evaluate the learning outcomes of the course and teachers' performance. Results A total of 35 medical workers from TARPH answered the baseline survey. The majority of the respondents had a college or higher degree. Up to 74.3% of the respondents never received any training in research before, and 42.9% of them had no experience of publishing peer-reviewed papers. After the DCR course, participants scored the course content and teachers with high to very high. 33.3%(ten) investigated participants completed their own clinical research proposal step by step as guided by the training course. All of the participants believed that it was necessary to continue the DCR course and open other research training courses. Conclusions Most medical staff at TARPH need the training and practical experience of research. The DCR course is an effective way to support their capacity building of scientific research.

Effect assessment of pathological professional training through the “Team-based” Medical Aid and Mentorship Program for Tibet

WANG Wen-ze, LUO Han-huan, ZHONG Ding-rong, SHI Jie, JIANG Ying, XIAO Yu, HUO Zhen, LIANG Zhi-yong

2022, 42(3):  537-540.  doi:10.16352/j.issn.1001-6325.2022.03.012

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“Team-based” Medical Aid and Mentorship Program for Tibet have been implemented for six years. The Department of Pathology at Peking Union Medical College Hospital (PUMCH) initiated a point-to-point aids to the Department of Pathology of Tibet Autonomous Region People's Hospital (TARPH) with six team members.With different teaching methods, the aids covered development of management regulations and technology training focusing on the capacity building of local hospital staff. The experts of PUMCH team contributed their wisdom and their experience to support training of local pathologists and improved their professional level. As a result, good social and economic benefits have been obtained.

Hospital Management

Application of membrane bioreactor treatment process in reconstruction of sewage station in general hospital

WANG Wen-ting, YANG De-yong, JIAO Jun-sheng

2022, 42(3):  541-544.  doi:10.16352/j.issn.1001-6325.2022.03.010

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Based on the space conditions and sewage treatment requirements of Peking Union Medical College Hospital, four sewage stations with first-grade treatment system were re-constructed using membrane bioreactor (MBR) treatment process. This paper reported the scheme selection, process flow and equipment selection. It shows that MBR treatment process is characterized by good effluent quality, compact equipment, small footprint and other advantages. It is worthy to recommend in other hospitals to support sewage station transformation.