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Table of Content

    05 July 2023, Volume 43 Issue 7
    Original Articles
    LncRNA RP11-686D22.10 is related with the survival of breast cancer patients, affects the proliferation, invasion of breast cancer cell lines
    TANG Yijun, WANG Wenlong, LI Jiangli, YANG Huan, SUN Jing
    2023, 43(7):  1023-1029.  doi:10.16352/j.issn.1001-6325.2023.07.1023
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    Objective To dientify the correlation between the expression of long-chain non-coding RNA (lncRNA) RP11-686D22.10 and the survival of breast cancer patients, and to explore the effect of over-expression of RP11-686D22.10 on the proliferation and invasion of breast cancer cell lines. Methods The relationship between the expression level of RP11-686D22.10 and the survival time of breast cancer patients was analyzed with GEPIA database. The expression of RP11-686D22.10 in mammary epithelial cell line MCF-10A and breast cancer cell lines MCF7, SKBR3, HCC1937, BT549 and MB-MDA-468 was detected by real-time quantitative polymerase chain reaction (RT-qPCR). SKBR3 cell with the least expression of RP11-686D22.10 was selected and transfected with control plasmid (NC group) and RP11-686D22.10 over-expression plasmid (RP11-686D22.10 group). The proliferation and invasion of breast cancer cells were detected by CCK-8 and Transwell assays, respectively. The Linc2Go database predicted the targeting microRNA (miRNA) of RP11-686D22.10, and the dual-luciferase reporter gene assay was used to target at relationship between RP11-686D22.10 and miR-454-3p. The expression of miR-454-3p was detected by RT-qPCR. Western blot was used to detect the protein expression of Wnt/β-catenin signaling pathway. Results Compared with breast cancer patients with low expression of RP11-686D22.10, patients with high expression of RP11-686D22.10 had longer overall survival (P<0.01) and longer disease-free survival (P<0.01). The expression of RP11-686D22.10 in MCF7, SKBR3, HCC1937, BT549, and MB-MDA-468 cells was lower than that in MCF-10A cell (P<0.05). The expression of RP11-686D22.10 in SKBR3 cells was the lowest (P<0.01). Compared with the NC group, the proliferation ability of SKBR3 cells in the RP11-686D22.10 group was decreased (P<0.05), and the cell invasion ability was decreased (P<0.01). RP11-686D22.10 could target and bind miR-454-3p (P<0.01). Compared with the NC group, the expression of miR-454-3p in the RP11-686D22.10 group was decreased (P<0.01), and the expressions of Wnt/β-catenin signaling pathway proteins CCND1, β-catenin, JUN, AXIN2, and MMP2 was decreased. Conclusions The expression level of RP11-686D22.10 is closely related to the survival of breast cancer patients. Over-expression of RP11-686D22.10 inhibits the proliferation and invasion of breast cancer cell lines by targeting at miR-454-3p.
    Baked licorice-derived sRNA alleviates lung injury mild ARDS mouse models possibly through targeting TNF-α
    JIANG Zhenyu, WANG Xiaona, TANG Kegong, CHEN Mingrui, JIANG Chengyu
    2023, 43(7):  1030-1039.  doi:10.16352/j.issn.1001-6325.2023.07.1030
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    Objective To explore the effect and mechanism of small RNA(sRNA)targeting at TNF-α in acute respiratory distress syndrome (ARDS). Methods Taking TNF-α as the target, screen sRNA from the sRNA database of Sini decoction by bioinformatics methods. The effect of sRNA targeting at TNF-α was verified by dual-luciferase reportergene system; the sRNA was transfected into the cells, then IL-1β, IL-6 and TNF-α of LPS or poly (I:C) induced A549 cells were detected by ELISA; the mRNA level of IL-1β, IL-6 and TNF-α in lipopolysaccharide(LPS) or poly(I:C) induced A549 cells was detected by RT-qPCR; Mild ARDS mouse models was developed by intratracheal instillation of LPS or poly(I:C) in mice. The mice were divided into native group, model group, NC group and sRNA-treated group. The concentration of IL-1β, IL-6 and TNF-α in BALF and serum was detected by ELISA. The survival of mild ARDS mouse models was observed. The survival rate of mice was counted. Results Bioinformatic analysis identified TNF-α mRNA binding 50 sRNAs and the dual-luciferase reporter gene system and THP1 inflammatory cell model identified the TNF-α-sRNA-9 that targeted at TNF-α, which was derived from Baked licorice in Sini decoction; Compared with NC group, TNF-α-sRNA-9 decreased the level of IL-1β, IL-6 and TNF-α in LPS or poly(I:C) induced A549 cells (P<0.05), and decreased the mRNA level (P<0.05). Compared with NC group, TNF-α-sRNA-9 treatment decreased the level of IL-1β, IL-6, and TNF-α in BALF and serum of mild ARDS mouse models induced by LPS or poly(I:C) (P<0.05) and increased the survival rate of mice. Conclusions TNFα-sRNA-9 alleviates lung injury in mild ARDS mouse models with potential mechanism of targeting at TNF-α.
    NOX4-mediated oxidative stress is involved in the occurrence of pituitary adenoma hemorrhagic stroke
    YANG Weizhu, LIU Zhong, ZHANG Fenglin, HUANG Yanlin, SUN Jin, TIAN Xinhua
    2023, 43(7):  1040-1044.  doi:10.16352/j.issn.1001-6325.2023.07.1040
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    Objective To study the role of reduced form of nicotinamide adenine dinucleotide phosphate(NADPH) oxidase 4 (NOX4) in pituitary adenoma hemorrhagic stroke(pituitary apoplexy,PA). Methods Specimens were collected from the remaining specimens of pituitary adenoma surgery in the Department of Neurosurgery, Zhongshan Hospital Affiliated to Xiamen University. According to postoperative pathology, the specimens were divided into hemorrhagic stroke(PA) group and non-stroke(non-PA) group, and 18 cases were selected from each group. Hematoxylin eosin (HE) staining was used to determine whether hemorrhage. The percentage of NOX4 positive cells was detected by immunohistochemical staining. The expression of NOX4 protein was detected by Western blot. DCFH-DA probe was used to detect the fluorescence intensity of reactive oxygen species (ROS). Apoptosis rate was detected by TUNEL. Results There was a large number of erythrocytes exuded between tissues in the hemorrhagic stroke group, but no red blood cell was found in the non-stroke group. The percentage of NOX4 positive cells in hemorrhagic stroke group (21.53%±4.25%) was significantly higher than that in non-stroke group (4.29%±1.71%) (P<0.001). The expression level of NOX4 in the hemorrhagic stroke group was significantly higher than that in the non-stroke group (P<0.001).The mean fluorescence intensity of ROS in hemorrhagic stroke group was significantly higher than that in non-stroke group(P<0.001). The percentage of apoptotic cells in hemorrhagic stroke group (38.64%±12.46%) was significantly higher than that in non-stroke group (6.19%±2.40%) (P<0.001). Conclusions NOX4 is involved in the occurrence of pituitary adenoma hemorrhagic stroke. NOX4-mediated oxidative stress regulation of apoptosis is found to be potential mechanism of hemorrhagic stroke in pituitary adenomas.
    Improving the generation of hepatocyte-like cells from pluripotent stem cells via high-efficient differentiation to definitive endoderm
    YAN Yucong, YANG Shuchun, JIA Yuyan, HUANG Yue
    2023, 43(7):  1045-1052.  doi:10.16352/j.issn.1001-6325.2023.07.1045
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    Objective To optimize the method of generating hepatocyte-like cells (HLCs) from pluripotent stem cells (PSCs)in vitro. Methods Considering the development of hepatocyte in vivo, a more efficient method was developed by optimizing biochemical factors to the stepwise differentiate PSCs into HLCs. Then the differentiation experiments were performed with human embryonic stem cell (hESC)line H9. The efficiency of the generation of HLCs was evaluated by morphological observation at different stages of differentiation. RT-qPCR and flow cytometry assay were used to examine the marker genes expression. Results A method of high-efficient hESCs differentiation to definitive endoderm (DE) was developed by adding cytokines activin A, bone morphogenetic protein 4(BMP4)and fibroblast growth factor 2(FGF2). It highly improved mRNA expression of marker genes at different stages. Venus fluorescence tracer and the results of flow-cytometry also showed that the population proportion of HLCs was significantly increased. Conclusions The high-efficient differentiation to DE improves differentiation efficiency of PSCs into HLCs.
    Interaction of family history of hypertension and overweight/obesity on hypertension in Han and Manchu adults in Hebei province
    JIANG Chenchen, CUI Ze, PAN Li, SUN Jixin, HE Huijing, HU Yaoda, SHAN Guangliang
    2023, 43(7):  1053-1059.  doi:10.16352/j.issn.1001-6325.2023.07.1053
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    Objective To investigate the effects of family history of hypertension, overweight/obesity and the interaction between the two factors on hypertension in Han and Manchu adults in Hebei Province, so to provide a reference for the prevention and control of hypertension in Hebei province. Methods A cross-section survey by questionnaires investigation and physical examination was conducted in Hebei province in July to August 2017, a multistage stratified cluster sampling covering 4 633 Han and Manchu people aged 20-80 years old in Hebei province was performed. A multivariate Logistic regression model was used to analyze the effects of family history of hypertension and overweight/obesity on hypertension. Results After adjusting for ethnicity, sex, age, geographic distribution(urban/rural), economic income,education, smoking, drinking, physical exercise and daily activity, the risk of hypertension was 3.6(95% CI: 2.51-5.18) times higher in those with a family history of hypertension than those without the family history. The risk of hypertension in overweight/obese people was 2.90(95% CI: 1.92-4.37) times higher than non-overweight/obese people. The risk of hypertension was 9.78(95% CI: 6.86-13.95) times higher in population with the factors of overweight plus family history as compared to those with neither factors.The relative excess risk due to interaction (RERI) was 4.28(95% CI: 2.59-5.96), the synergy index (SI) was 1.95(95% CI: 1.54-2.47) and the attributable proportion due to interaction (AP) was 0.44(95% CI: 0.33-0.54). Conclusions Both family history of hypertension and overweight/obesity are associated with an increased risk of hypertension, and there is a positive additive interaction between the two factors.
    STAT1-induced LINC00987 regulates miR-223-3p/FZD4 and promotes M2-type polarization of human glioblastoma-associated macrophages
    ZHANG Baorui, MA Chunxiao, QIN Lijie
    2023, 43(7):  1060-1068.  doi:10.16352/j.issn.1001-6325.2023.07.1060
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    Objective To investigate the effects of LINC00987 activated by STAT1 on M2-type polarization of human glioblastoma (GBM) associated macrophages. Methods LncRNAs with dysregulated expression in GBM were screened by GEO analysis. The expression of LINC00987, miR-223-3p and FZD4 was detected by RT-qPCR. THP-1 cells were induced into M0, M1, and M2 macrophages, and GBM cells were co-cultured with THP-1 cells. The proportion of CD14+/CD206+ macrophages was detected by flow cytometry, The expression of M2-type macrophage-related factors (VEGF, TGF-β1) was detected by ELISA. Over-expression of LINC00987 promoted the M2-type polarization of GBM-related macrophages (P<0.05). FZD4 knockdown inhibited M2-type polarization of GBM-related macrophages, but this effect was partially reversed by over expression of LINC00987(P<0.05). Results Compared with paracancerous tissues and HEB cells, the expressions of LINC00987 and FZD4 in GBM tissues and cells were up-regulated while the expression of miR-223-3p was down-regulated(P<0.05). In GBM cells, STAT1 activated LINC00987 and affected miR-223-3p/FZD4 axis. LINC00987 knockdown inhibited M2-type polarization of GBM-related macrophages, but this effect was partially reversed by miR-223-3p inhibitors(P<0.05). Conclusions STAT1 activated LINC00987 regulates miR-223-3p/FZD4 axis to induce M2-type polarization of GBM-related macrophages.
    Effects of kaempferol regulating VPS9D1-AS1/miR-377-3p on proliferation and apoptosis of cervical cancer cell line SiHa
    ZHANG Yuxin, ZHU Ziwei, ZHANG Yan, HU Yuhong
    2023, 43(7):  1069-1078.  doi:10.16352/j.issn.1001-6325.2023.07.1069
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    Objective To investigate the effect of kaempferol on regulation of long non-coding RNA(lncRNA) vps9d1 antisense RNA 1 (VPS9D1-AS1) /miR-377-3p and on biological behavior of cervical cancer cells. Methods The cervical cancer cell line SiHa was divided into control group, low, medium and high dose of kaempferol group (5 μmol/L、10 μmol/L and 20 μmol/L kaempferol), si-NC group (transfected si-NC), si-VPS9D1-AS1 group (transfected si-VPS9D1-AS1), kaempferol + pcDNA-VPS9D1-AS1 group (20 mg/L kaempferol+transfected pcDNA-VPS9D1-AS1), kaempferol + anti-miR-377-3p group (20 mg/L kaempferol + transfected anti-miR-377-3p).VPS9D1-AS1, miR-377-3p expression, cell activity and colony formation were measured by RT-qPCR, MTT assay and colony formation assay; Cell apoptosis, migration, invasion and Bax/Bcl-2 expression were measured by flow cytometry, scratch test, Transwell test and Western blot. Luciferase activity assay was used to evaluate the targeting effect of VPS9D1-AS1 on miR-377-3p. Results Compared with the control, low, medium and high doses of kaempferol decreased the expression level of VPS9D1-AS1, the number of colony formation, cell activity, Bcl-2 protein expression level, migration distance and the number of invasive cells in SiHa cells, and increased the expression level of miR-377-3p, apoptosis rate as well as Bax protein expression level in a concentration dependent manner (P<0.05). After VPS9D1-AS1 was silenced in SiHa cells, the expression level of miR-377-3p, apoptosis rate and Bax protein expression level were higher than those in si-NC group; The colony forming number, Bcl-2 protein expression, cell activity, migration distance and the accounting of invasive cells in si-NC group were significantly(P<0.05) lower than those in si-NC group. VPS9D1-AS1 targeted at miR-377-3p. The effects of kaempferol treated SiHa on proliferation, apoptosis, migration and invasion were reversed by over-expression of VPS9D1-AS1 or inhibition of miR-377-3p. Conclusions Kaempferol can inhibit the proliferation, migration and invasion of cervical cancer cells and accelerate apoptosis by down regulating VPS9D1-AS1 targeted at miR-377-3p.
    High expression of hsa_circ_0017650 in human adipocyte differentiation process and adipose tissue of obese people
    ZHANG Jing, WU Kai, HUANG Wei
    2023, 43(7):  1079-1083.  doi:10.16352/j.issn.1001-6325.2023.07.1079
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    Objective To investigate the expression changes of hsa_circ_0017650 in the human adipocyte differentiation process and adipose tissue of obese people. Methods The changes of hsa_circ_0017650 on the 0th, 1st, 3rd, 5th, 7th, 9th and 12th days of human preadipocyte differentiation were determined by RT-qPCR; Oil Red O staining was performed to observe the formation of lipid droplets on 0th, 6th, and 12th days of human preadipocyte differentiation. The mRNA levels of CCAAT enhancer-binding protein α (C/EBPα), fatty acid-binding protein 4 (FABP4/ap2), and lipoprotein lipase (LPL) in human preadipocytes were determined by RT-qPCR to identify the differentiation of human preadipocytes. A total of 125 patients who underwent appendectomy in our hospital were selected as research subjects, and were divided into obesity group (53 cases) and control group (72 cases) according to body mass index (BMI). The expression differences of hsa_circ_0017650 in retroperitoneal white adipose tissue of the two groups were compared. The Pearson’s correlation coefficient between the expression level of hsa_circ_0017650 and BMI, triacylglycerol (TG), total cholesterol (TC), low density lipoprotein cholesterol (LDL-C) and high density lipoprotein cholesterol (HDL-C) in obesity group was analyzed. Results Human preadipocytes gradually changed from spindle to oval or round, with a gradual increase in volume and an increase in fat droplets. On the 12th day of culture the levels of C / EBPα, FABP4/ap2 and LPL in human precursor adipocytes were higher than those on day 0 (P<0.001). Compared with baseline of day zero(1.02±0.13), the expression of hsa_circ_0017650 increased on the 1st (2.47±0.30), 3rd (27.38±3.42), 5th (15.62±1.95), 7th (16.24±2.03), 9th (18.37±2.29) and 12th (16.24±2.21), and reached the peak on the 3rd day (P<0.001). Compared with the control group, the levels of BMI, waist-to-hip ratio, TC, TG, LDL-C and of hsa_circ_0017650 in white adipose tissue increased in the obesity group (P<0.001); Hsa_circ_0017650 was positively correlated with BMI, TG, and LDL-C (r=0.554, 0.569, 0.618, P<0.001). Conclusions Hsa_circ_0017650 is highly expressed during human adipocyte differentiation and mainly in white adipose tissue of obese individual.It is closely related to obesity and is potential for the treatment of obese patients.
    miR-199-3p inhibits diabetes retinopathy in rats by targeting at VegfA
    ZHOU Yue, LIN Jingna
    2023, 43(7):  1084-1089.  doi:10.16352/j.issn.1001-6325.2023.07.1084
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    Objective To study the role and mechanism of mir-199-3p in the pathogenesis of diabetic retinopathy (DR). Methods The diabetic model of rats was established by injection of streptozotocin. The expression of miR-199-3p and vascular endothelial growth factor A (VEGFA) in the retina of rats was detected by RT-qPCR. The targeting relationship between miR-199-3p and VegfA in retinal cells was studied by dual luciferase assays. Apoptosis of retinal cells was evaluated by TUNEL staining. Protein of VEGFA and transforming growth factor-β(TGF-β), Hepatocyte growth factor (HGF), pigment epithelium derived factor (PEDF) were detected by Western blot. Results The retinal staining showed that the pathological changes such as retinal capillary dilation and vascular curvature in diabetic rats. The results of reverse transcription showed that the expression of miR-199-3p decreased and VEGFA increased in the retina of diabetic rats significantly. The targeting relationship between miR-199-3p and VegfA was confirmed by dual luciferase assays. The low-expression of miR-199-3p significantly reduced the number of retinal ganglion cells and enhance the apoptosis of retinal cells, while the low expression-miR-199-3p and VEGFA significantly reversed the effects of miR-199-3p on retinal cells(P<0.01). Western blot results showed that the expression of TGF-β and HGF decreased significantly after over-expression of miR-199-3p with the increased expression of PEDF(P<0.001). However, over-expression of miR-199-3p combined with VEGFA expression significantly reversed the expression of these proteins. Conclusions miR-199-3p inhibits diabetic retinopathy by targeting at VegfA.
    Expression and clinical significance of SIRT2 in urothelial bladder cancer and its impact on cancer cell proliferation, invasion and migration
    JIN Xiaoxia, LU Xiaoyun, LIU Yushan, LIANG Lina
    2023, 43(7):  1090-1096.  doi:10.16352/j.issn.1001-6325.2023.07.1090
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    Objective To investigate the expression and clinical significance of silent information regulator 2 (SIRT2) form tissue of urothelial bladder cancer(UBC) and the effects of down-regulation of SIRT2 on the proliferation, invasion and migration of bladder cancer cells. Methods The expression of SIRT2 protein was detected in 95 samples of urothelial bladder cancer tissues and 39 paracancerous tissues using the immunohistochemical EnVision method. Its relationship with clinicopathological parameters was analyzed. The expression of SIRT2 mRNA in bladder cancer cell lines was detected by RT-qPCR, and the expression levels of SIRT2 protein in bladder cancer cell lines, 12 pairs of fresh bladder cancer tissues were detected by Western blot. Lentiviral infection was used to stably downregulate the SIRT2 gene in bladder cancer cell line T24. CCK8 and cell colony assays were performed to detect changes in cell proliferation ability and Transwell assay to detect changes in cell invasion and migration activity. Results The positive expression rate of SIRT2 in urothelial bladder cancer tissues was 84%(80/95), which was higher than that in normal uroepithelial tissues (5%, 2/39) (P<0.05). The expression rate of SIRT2 was higher in the high-grade uroepithelial cancer group (95%, 38/40) than in the low-grade group (76%, 42/55) (P<0.05); The expression rate was higher in the muscle-infiltrated group (96%, 24/25) than in the non-muscle-infiltrated group (80%, 56/70) (P<0.05). Expression of SIRT2 protein was higher in fresh bladder cancer tissues than in normal tissues adjacent to the cancer. Expression of mRNA and protein of SIRT2 were also upregulated in bladder cancer cell lines. Knockdown of SIRT2 in T24 cells resulted in a significant decrease in cell activity of proliferation, invasion and migration (P<0.05). Conclusions SIRT2 is highly expressed in bladder uroepithelial carcinoma, and interfers with SIRT2 significantly and so inhibites bladder cancer cell proliferation, invasion and migration.
    Comparison of the expression of Smad1 in renal tissues from patients with diabetic nephropathy and other types of renal diseases
    CHEN Xianxian, FENG Lie, ZHOU Ying, DONG Xiangnan
    2023, 43(7):  1097-1103.  doi:10.16352/j.issn.1001-6325.2023.07.1097
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    Objective To compare urinary Smad1 and the protein content of Smad1 in renal tissues of patients with diabetic nephropathy (DN) and the patients with other types of nephropathy. Methods According to the result of renal biopsy, the patients were collected and divided into four groups as follows: the patients with diabetes nephropathy(DN group, 9 cases), with focal segmental glomerulosclerosis (FSGS group, 5 cases), with IgA nephropathy (IgA group, 12 cases) and with minimal change nephrosis (MNP group, 10 cases). Urinary Smad1 was examined using ELISA. The expression of Smad1, Col4 and α-SMA in renal tissues was detected by immunohistochemistry and immunofluorescence staining method. Results Urinary Smad1 can be detected in the DN group and there other groups (FSGS group, IgA group and MNP group). The urinary Smad1 from four groups was higher significantly than those from 47 healthy people with routine physical examination (P<0.05). The expression of Smad1 in different area of the renal tissues(the area of glomerulus and renal tubule) can be observed by immuno-histochemistry staining in the patients with four different types of nephropathy. The Smad1 expression in the renal tissue of the patients with diabetes nephropathy was higher than that of three other groups (P<0.05). The co-expression of Col4 and α-SMA with Smad1 in different area of the renal tissues (the area of glomerulus and renal tubule) was found. Conclusions The over-expression of Smad1 in different area of the renal tissues can be observed and is related with renal damage.
    Down-regulation of BECN1 inhibits the proliferation, invasion and migration of human leukemia cell line THP-1
    ZENG Yankun, HONG Xiaoying, CHEN Qinglin
    2023, 43(7):  1104-1109.  doi:10.16352/j.issn.1001-6325.2023.07.1104
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    Objective To analyze the effect of down-regulation of BECN1 on the proliferation,invasion and migration of human leukemia cell line THP-1,and to explore its potential molecular mechanism. Methods The lentivirus expressing shRNA targeting at BECN1 and the negative control were constructed by RNA interference. THP-1 cells were divided into control group(control),negative control group(NC) and BECN1 interference group (BECN1-shRNA). Stable cell strains were selected by puromycin after infection by lentivirus. The down-regulation effect of BECN1 was identified by RT-qPCR and Western blot. MTT and Transwell assays were used respectively to detect the proliferation,invasion and migration of THP-1 cells. The expression of MMP 2 and MMP 9 were detected by Western blot. Results THP-1 stable cell strain expressing GFP was obtained by puromycin selection. Compared with the control and negative control group, the BECN1 mRNA and protein expression in the BECN1-shRNA group were significantly decreased (P<0.05); The proliferation,invasion and migration of cells also decreased significantly(P<0.05);The expression of MMP 2 and MMP 9 decreased (P<0.05). Conclusions Down-regulation of BECN1 may inhibit the proliferation,invasion and migration of THP-1 cells,and its molecular mechanism is related to the down-regulation of MMP 2 and MMP 9.
    Mesenchymal stem cell-engineered DNA nanostructures and the application in targeted delivery to the lungs of mice
    LI Zhuoting, SHANG Yingxu, WANG Haiyan, JIANG Qiao, DING Baoquan, LI Jing, ZHAO Chunhua
    2023, 43(7):  1110-1116.  doi:10.16352/j.issn.1001-6325.2023.07.1110
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    Objective Modifying 3D tubular DNA origami nanostructures (DONs) onto the mesenchymal stem cells (MSCs) membrane surface to obtain DONs@MSCs. Utilizing the characteristics of MSCs homing lung tissue to achieve targeted delivery of DONs to the lungs of mice. Methods The computer-aided tools for DNA sequence were used to design and obtain obtain pre-designed DONs. The morphology of assembled DONs was characterized by atomic force microscopy(AFM), and the upload of functional groups was verified by agarose gel electrophoresis (AGE). Fluorescent labeled single-strand DNA(ssDNA) was modified on the surface of MSCs using biological orthometabolic sugar engineering. According to the principle of complementary base pairing, DONs were modified on the MSCs surface. Using confocal microscopy and flow cytometry to detect the ssDNA and DONs, and using in vivo imaging to observe the distribution of DONs in lung tissue. Results DONs had been successfully assembled. MSCs membrane surface was successfully modified with ssDNA, and DONs were successfully hybridized to obtain DONs@MSCs. Compared to DONs, the enrichment of DONs@MSCs in the lungs of mice was significantly enhanced. Conclusions DONs@MSCs can be used for DONs targeted delivery to lung tissue of mice. This method provides a new strategy for drug delivery of DONs in lung tissues.
    Expression of peptidylarginine deiminase 2 and 4 in malignant mesothelioma and its clinical significance
    GAN Yihan, SHEN Wei, CHEN Yitong, YAN Kaili, HU Shuaiyue, JIANG Zhaoqiang, YING Shibo
    2023, 43(7):  1117-1121.  doi:10.16352/j.issn.1001-6325.2023.07.1117
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    Objective To investigate the expression of peptidylarginine deiminase 2 and 4 (PAD2 and PAD4) in malignant mesothelioma (MM), their correlation with citrullinated proteins and clinical significance. Methods The pathological samples of 45 cases of MM in Zhejiang province from 2014 to 2021 were collected retrospectively for constructing paraffin-embedded tissue microarray. The expressions of PAD2 and PAD4 in tumor and paired adjacent tissues were detected by immunohistochemistry and the clinical significance was analyzed by utilizing bioinformatics database. Results The expressions of PAD2 and PAD4 in MM tissues were significantly higher than those in adjacent tissues (P<0.05). The protein expression of PAD2 and PAD4 was positively correlated with epithelial mesothelioma, peritoneal mesothelioma and pleural mesothelioma (P<0.05). There was a positive correlation between PAD2 and PAD4 mRNA transcription level (P<0.05). The expressions of PAD2 and PAD4 were positively correlated with citrullinated proteins, respectively (P<0.05). The overall survival(OS) of PAD2 and PAD4 high expression group was shorter than that of low expression group, the progression free interval(PFI) of PAD4 high expression group was longer than that of low expression group (P<0.05). Conclusions PAD2 and PAD4 are potentially involved in the progression and prognosis of MM as biomarkers or therapeutic targets.
    Total flavonoids of Hippophae rhamnoides L. inhibits proliferation of bladder cancer cell line T24
    ZHANG Peibo, LI Yi
    2023, 43(7):  1122-1126.  doi:10.16352/j.issn.1001-6325.2023.07.1122
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    Objective To investigate the effect of total flavonoids of Hippophae rhamnoides L.(HTF) on the proliferation and apoptosis of bladder cancer cell line T24 and underlying mechanism. Methods The bladder cancer T24 cells were incubated with different concentrations (25, 50, 100 μg/mL) of HTF for 24 h, then CCK-8 assay, flow cytometry and Western blot were used to detect proliferation, apoptosis and the expression of Bcl-2 and Bax proteins, respectively. The expression of miR-144 was detected by RT-qPCR. miR-144 inhibitors were transfected into T24 cells to construct T24 cells whose expression of miR-144 was knocked down,then 100 μg/mL HTF was used to interfere with the miR-144-knoched down T24 cells. The same methods were used to detect the effects of knocking down of miR-144 on cell proliferation and apoptosis of T24 cell. Results Compared with the control group, different concentrations of HTF reduced the cell A value and the expression of Bcl-2, and increased the apoptosis rate and the expression of Bax. At the same time, the expression of miR-144 (P<0.05) was increased.The A value of T24 cells that knocked down miR-144 and the expression of Bcl-2 protein in the cells was higher than those of control T24 cells(P<0.05), but the apoptosis rate and the expression of Bax protein were lower(P<0.05). After T24 cells that knocked down miR-144 were treated with 100 μg/mL HTF, their A value and the expression of Bcl-2 protein in the cells were higher than those of T24 cells that not knocked down miR-144(P<0.05), but the apoptosis rate and the expression of Bax protein were lower than those of control T24 cells (P<0.05). Conclusions HTF may slow-down the proliferation of bladder cancer cells and speed-up their apoptosis by up-regulating miR-144.
    Construction and validation of a risk prediction model for frailty in patients with inflammatory bowel disease
    YOU Lili, YANG Lixiao, WANG Siyao, QIAN Na, LU Min, SONG Meng, GUAN Yuxia
    2023, 43(7):  1127-1133.  doi:10.16352/j.issn.1001-6325.2023.07.1127
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    Objective To establish a risk prediction model for frailty in patients with inflammatory bowel disease and verify its prediction effect. Methods A classic sampling method was used to collect samples from 360 patients with inflammatory bowel disease who were hospitalized in the Department of Gastroenterology of Peking Union Medical College Hospital from July 2021 to November 2021. According to the evaluation results of the FRAIL Scale (FS), patients were divided into two group: frail group and non-frail group. The differences of demographic factors and disease related factors between the two categories of patients were compared. Randomized number method was used to divide the model group and verification group at a ratio of 8∶2. Logistic regression analysis was used to establish a risk prediction model. Hosmer-Lemeshow χ2 was used to test the prediction model fitting effect, and receiver operator characteristic curve (ROC curve) was drawn. Area under ROC curve (AUC) and Youden index were calculated to evaluate the differentiation ability of the prediction model. Results The incidence of frailty in patients with inflammatory bowel disease was 16.6%. Age (OR=1.041), emaciation (OR=10.135), several times of exercise per month (OR=0.345), ulcerative colitis (OR=2.952), quality of life (OR=0.979), family care index (OR=1.629), depression (OR=1.194) and sleep disorders (OR=1.158) were independent influencing factors (P<0.05). The results of Hosmer-Lemeshow test showed that χ2=1.594, P=0.991, AUC=0.934 (95% confidence interval, 95% CI: 0.901~0.966), the maximum Youden index was 0.728, the optimal critical value was 0.128, the sensitivity was 92.0%, and the specificity was 81.8%. The validation results show that the sensitivity of the model is 88.9%, the specificity is 72.6%, and the accuracy is 91.55%. Conclusions The prediction effect of the risk prediction model is good, which can provide a reference for clinical evaluation of the frail risk of patients with inflammatory bowel disease.
    Clinical Sciences
    Clinical features and neonatal outcomes of ten pregnant women infected with Omicron variant of SARS-CoV-2
    BAO Qiufang, CAI Qiongyi, YANG Huijuan, HONG Linliang
    2023, 43(7):  1134-1137.  doi:10.16352/j.issn.1001-6325.2023.07.1134
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    Objective To investigate the clinical characteristics and neonatal outcomes of pregnant women infected with Omicron variant of SARS-CoV-2 in order to provide reference for diagnosis and treatment. Methods Data of 10 pregnant women infected with Omicron on variant in Quanzhou First Hospital were collected, their epidemiological and clinical characteristics as well as the growth and development of newborns were analyzed. Results All the 10 patients had a history of exposure to the epidemic area and showed mild or asymptomatic clinical symptoms. The first clinical symptoms were fever and cough. All pregnancies and the delivery was ended smoothly with no adverse outcome. Conclusions Infection with the Omicron variant of SARS-CoV-2 during pregnancy has little effect on pregnancy outcome but this result needs to be confirmed by a larger sample study.
    Perioperative management of adult patients with congenital heart disease undergoing neurosurgery
    WU Bei, YANG Liu, HAN Ruquan
    2023, 43(7):  1138-1142.  doi:10.16352/j.issn.1001-6325.2023.07.1138
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    Objective To review and analyze the perioperative anesthesia management of patients with adult congenital heart disease (ACHD) underwent neurosurgery, and to summarize the key points of perioperative anesthesia management for these patients, so as to achieve the goal of high clinical quality development. Methods This retrospective study analyzed the perioperative medical records of patients with ACHD underwent neurosurgery from January 2018 to December 2022. Demographic data, CHD types and complications, perioperative anesthesia management, postoperative complications and prognosis were analyzed. Results A total of 21 ACHD patients underwent neurosurgery were included in this analysis, 16 with atrial septal defect, 4 with ventricular septal defect, and 1 with single atrium and single ventricle accompanied with cyanosis and compensated hyper-hemoglobin. Left to right shunt was observed in 19 cases, and bidirectional shunt in 1 case. 11 cases were associated with heart chamber enlargement(right heart and left atrium). 20 cases received general anesthesia, and 1 case received nerve block regional anesthesia. six cases were persistently injected vasoactive drugs to maintain circulation stabilization. The intraoperative fluid balance was -6~+ 913 mL/h. The operation time was 46~355 min. 6 cases returned to ICU after surgery. 4 cases suffered central nervous system infection, 2 electrolytes disturbances, 2 coagulation abnormalities, 1 lower limb venous thrombosis, 1 abnormal liver function, and 1 death. The average length of hospitalization was (12.5±5.9) days. Conclusions Circulatory,cardiopulmonary function and fluid therapy are the key points of perioperative management of ACHD patients undergoing neurosurgery.
    Mini Reviews
    Progress on the pathological roles of immune checkpoint CD276 protein in tumorigenesis and development
    CHEN Zhongjie, HUANG Jiaru, HU Qing, ZHAO Zhenqin, CHEN Qiaoqiao, SUN Junyuan, JIA Jing
    2023, 43(7):  1143-1147.  doi:10.16352/j.issn.1001-6325.2023.07.1143
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    The immune checkpoint CD276 protein, belonging to the B7 family molecules, could interact with important signaling pathways, such as phosphatidylinositol 3 kinase/protein kinase B(PI3K/AKT)signaling pathway, resulting to the regulation of tumor energy metabolism, inhibition of the anti-tumor activity of immune cells, promotion of tumor angiogenesis, and the assistance of tumor immune escape and metastasis. CD276 also potentially related to tumor stemness and chemotherapy resistance closely. Several antibody drugs and cell therapy methods targeting at CD276 have been proved in clinical trials. CD276 can also be used as a biomarker for tumor diagnosis and subtyping. The upstream regulatory mechanisms of CD276 include transcriptional level regulation mediated by non-coding RNA and transcription factors, and post-translational modification level regulation mediated by methylation and glycosylation.
    An update on the mechanism of neural stem cell transplantation in the treatment of Alzheimer’s disease
    CHEN Xiaokun, BAO Xinjie, GAO Jun, LI Yongning
    2023, 43(7):  1148-1151.  doi:10.16352/j.issn.1001-6325.2023.07.1148
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    At present, no effective strategy exists to slow down the progression of Alzheimer’s disease (AD). A deeper understanding of the pathogenesis of AD and the development of new therapeutic approaches are essential. Stem cell transplantation for the treatment of neurodegenerative diseases is currently a research hotspot. Numerous studies have suggested that neural stem cells (NSCs) can differentiate into functional neurons, replace dead neurons, and form synaptic connections with adjacent surviving neurons, thereby achieving a therapeutic effect. In theory, the use of NSCs to replace and restore damaged cholinergic neurons and synapses may provide a new therapeutic option for AD. This review summarizes the recent preclinical research progress of NSC transplantation for the treatment of AD.
    Progress of specialized pro-resolving mediators in diabetic kidney disease
    LIU Chong, LIU Yingying, GUO Zhaoan
    2023, 43(7):  1152-1156.  doi:10.16352/j.issn.1001-6325.2023.07.1152
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    Specialized pro-resolving mediators (SPMs) can promote the regression of chronic inflammatory response in diabetic kidney disease (DKD), inhibit renal fibrosis and protect renal function by inhibiting renal intrinsic cell inflammatory response, antioxidant stress injury and regulate macrophage transformation. With the in-depth study of SPMs, it is expected to become a new target for the treatment of chronic inflammatory diseases such as DKD.
    Role of cigarette smoke in the development of severe pneumonia and related research progress
    ZENG Xin, ZHOU Qifeng, TONG Jin
    2023, 43(7):  1157-1161.  doi:10.16352/j.issn.1001-6325.2023.07.1157
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    Cigarette smoke(CS) contains a large number of toxic and carcinogenic compounds. CS induces severe lung infections through different molecular and cellular mechanisms of alteration of respiratory tract structure, innate and adaptive immunity and microbiome homeostasis. CS is strongly associated with poor prognosis in corona virus disease 2019(COVID-19). The different mechanisms through which CS effects contribute to the understanding of the pathophysiology of CS-induced severe pneumonia and explore potential medicine.
    Progress of NLRP3 inflammasome in the pathogenesis of heart disease
    GAO Yuan, ZHENG Gang, QI Jing, ZHANG Feng
    2023, 43(7):  1162-1166.  doi:10.16352/j.issn.1001-6325.2023.07.1162
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    NLRP3 inflammatsome is the most characteristic inflammasome. It can play some roles in the pathogenesis of heart diseases such as heart failure, myocardial infarction, atherosclerosis and arrhythmia. Inhibiting the activation of NLRP3 inflammasome can effectively treat heart diseases. The assembly and activation process of inflammasome are complex. Revealing its activation process and its relationship with heart disease may develop new drugs targeting at the prevention and treatment of heart disease.
    Advances on the role and molecular mechanism of microRNAs in cardiac hypertrophy
    WANG Fan, DENG Yongzhi
    2023, 43(7):  1167-1170.  doi:10.16352/j.issn.1001-6325.2023.07.1167
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    MicroRNAs(miRNAs) are a class of non-coding RNAs associated with a variety of cardiovascular diseases, including hypertension, heart failure and myocardial infarction. Expression of miRNAs changes during cardiac hypertrophy, which regulates cardiac hypertrophy by regulating cardiomyocyte autophagy, inflammatory response and energy metabolism. This article reviews the expression changes, potential targets and molecular mechanisms of miRNAs in cardiac hypertrophy.
    Medical Education
    Teaching reform of integrated course of endocrine system in basic medicine
    LI Wei, ZHU Huijuan, XIA Weibo, ZHU Lei
    2023, 43(7):  1171-1174.  doi:10.16352/j.issn.1001-6325.2023.07.1171
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    The 4+4 Medical Doctor Program set up by Peking Union Medical College has the characteristics of diverse professional backgrounds and short learning experience of medical courses. It is a pilot project to set up an integrated course of “organ-system-function”(OSF) basic medicine in the program. Teachers try to build an autonomous learning environment through the modular object-oriented dynamic learning environment(Moodle) platform in endocrine system teaching, carry out collective lesson preparation and trial lecture system, combine with the history of medicine and the introduction to medical scientific research, in order to explore a teaching model suitable for 4+4 Medical Doctor Program. According to the questionnaire survey, 80.0% of the students are generally satisfied with the teaching effect, 45.0% suggest that the basic knowledge content should be strengthened, and 55.0% of them believe that it is necessary to increase the humanistic knowledge related to the professional content.
    Cultivation of medical student’s science popularization ability in teaching of pharmacy
    ZHANG Ying, FANG Hongjuan
    2023, 43(7):  1175-1178.  doi:10.16352/j.issn.1001-6325.2023.07.1175
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    The scientific popularization of pharmacy knowledge is one of the important directions for the transformation of pharmaceutical services in China. As the main force of the future pharmaceutical popular science information dissemination, the capacity building of popular science of college students is crucial. This article discusses the necessity and significance of improving the capacity of pharmacy college students in popular science based on current situation and gives some suggestions on how to integrate the education in medical colleges, clinical pharmacists and pharmacy colleges to improve the pharmacy college students’ popular science capacity. The recommended strategy is to strengthen the cultivation of students’ popular science awareness and sense of social responsibility, makes them to be competent in pharmaceutical popular science education and serves the general public in their future work to build a “Healthy China”.