Table of Content

05 January 2024, Volume 44 Issue 1

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Original Articles

Circ-DDX5 inhibits the proliferation and invasion of human breast cancer cell lines by targeting miR-3940

LI Jiangli, SUN Jing, TANG Yijun, GUO Junlan, CHEN Bo, GUO Shengnan

2024, 44(1):  1-7.  doi:10.16352/j.issn.1001-6325.2024.01.0001

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Objective To investigate the expression of circular-RNA DDX5(circ-DDX5) in breast cancer tissues and its relationship with the clinical stage of breast cancer patients, and to analyze the regulatory mechanism of circ-DDX5 on the proliferation and invasion of human breast cancer cell line. Methods The expression level of circ-DDX5 in breast cancer tissues and its correlation with the clinical stage of breast cancer patients were analyzed by TCGA database. Bioinformatics analysis and dual-luciferase reporter gene experiments verified the targeting relationship between circ-DDX5 and miR-3940. The correlation between circ-DDX5 and miR-3940 expression in breast cancer tissues was analyzed by TCGA database. The expression level of circ-DDX5 in breast cancer SK-BR-3, MDA-MB-231, BT-549, MCF-7, and HCC-1937 cells was detected by RT-qPCR. The circ-DDX5 over-expression plasmid and negative control plasmid were transfected into MDA-MB-231 cells, which were named circ-DDX5 group and NC group, respectively. The proliferation and invasion of MDA-MB-231 cells in the circ-DDX5 group and the NC group were detected by colony formation assay and Transwell assay. The expressions of proliferation phenotype protein and invasion phenotype protein of MDA-MB-231 cells were detected by Western blot. The expression level of miR-3940 in MDA-MB-231 cells of circ-DDX5 group and NC group was detected by RT-qPCR. Results The expression of circ-DDX5 in breast cancer tissues was lower than that in adjacent tissues (P＜0.01) and the expression level of circ-DDX5 was negatively correlated with the clinical stage of breast cancer patients (P＜0.01). There was a targeting relationship between circ-DDX5 and miR-3940 (P＜0.01). The expression of circ-DDX5 and miR-3940 in breast cancer tissue was negatively correlated (P＜0.01). The expression of circ-DDX5 in human breast cancer cell lines was lower than that in immortalized breast epithelial cells MCF-10A (P＜0.05 or P＜0.01). Compared with the NC group, the over-expression of circ-DDX5 could significantly inhibit the proliferation and invasion of MDA-MB-231 cells(P＜0.01), as well as the proliferation phenotype proteins (cyclin C, CDK3) and invasion phenotype proteins (Snail, vimentin) expression (P＜0.01) and miR-3940 expression (P＜0.01). Conclusions The expression of circ-DDX5 in breast cancer tissues and cells is low. circ-DDX5 inhibits the proliferation and invasion of breast cancer MDA-MB-231 cells by targeting the expression of miR-3940.

miR-139-5p enhances the inhibition effect of metformin on cell proliferation of pancreatic cancer cell line PANC-1 cultured in normal-glucose medium

YU Jie, MA Minglei, ZHANG Huabing, PING Fan, LI Wei, XU Lingling, LI Yuxiu

2024, 44(1):  8-15.  doi:10.16352/j.issn.1001-6325.2024.01.0008

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Objective To investigate the effects of metformin(Met) on the proliferation of pancreatic cancer cells under different glucose concentration culture conditions, and to find the potential role of miR-139-5p in the process. Methods PANC-1 cells were treated with different concentrations of metformin(0/5/10/20 mmol/L) in 25 mmol/L(high-glucose group,HG) or 5 mmol/L(normal-glucose group,NG) glucose culture, cell proliferation, apoptosis, migration and cell cycle were detected after 48 h. The expression of miR-139-5p was quantitatively detected by RT-qPCR, and the miR-139-5p mimics were transfected into PANC-1 cells to clarify the role of miR-139-5p. Results Metformin inhibited the proliferation, promoted apoptosis, and induced S phase and G₂/M phase arrest of PANC-1 cells under in high glucose and normal glucose culture conditions, and its anti-proliferation and pro-apoptosis effects were more significant in the normal glucose groups. The expression of miR-139-5p was up-regulated by metformin treatment in normal but not in high glucose culture. Further studies showed that miR-139-5p mimics inhibited of PANC-1 cells proliferation without metformin pre-incubation and enhanced the anti-proliferation effect of 5 mmol/L metformin. The pro-apoptotic effect of 10 mmol/L metformin in normal glucose culture conditions. Conclusions In normal-glucose culture conditions, metformin can inhibit proliferation, induce apoptosis and cell cycle arrest of PANC-1 cells more significantly than in higher-glucose culture, which may be partly related to the up-regulation of miR-139-5p.

Culture of glioblastoma U87 stem-like cells and identification of its metabolic phenotype and tumorigenic ability

QIU Jiaxing, LIU Yuhan, GUO Hongjiang, ZHANG Diya, WANG Yucheng, JU Rui, GUO Lei

2024, 44(1):  16-22.  doi:10.16352/j.issn.1001-6325.2024.01.0016

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Objective To cultivate glioblastoma U87 stem-like cells (SLCs) and to detect the level of stemness biomarkers, mitochondrial respiratory capacity and the capacity of in vivo tumorigenesis. Methods B-27, growth factors EGF and bFGF was added into DMEM/F-12 culture in serum-free stem cell culture medium for U87 SLCs. Suspended culture of U87 SLCs was suspended using the neuro-sphere formation assay, while adherent culture of U87 SLCs was achieved by coating Matrigel matrix on the culture surface. The mRNA and protein level of stemness biomarkers in culture were detected using real-time quantitative PCR and Western blot. The proportion of CD133⁺ cells in culture was detected by flow cytometry. The changes of cell oxygen consumption rate were detected by Seahorse cell metabolism analysis. Cell tumorigenesis ability was verified by subcutaneous tumor transplantation in animals. Results U87 SLCs in stem cell culture medium would grow into typical sphere morphology within one week, and the spheres would continue to grow as the culture process prolongs. At the appropriate concentration of adhesive, U87 SLCs adhered to and grow well in stem cell culture medium. The mRNA transcription of stemness biomarkers such as CD133, nestin, OLIG2, CD44, CD15, and integrin α6(ITGA6) was significantly increased as found in both culture methods, and the protein levels of CD133 and nestin were also increased under both methods(P＜0.05). U87 SLCs showed higher mitochondrial reserve respiratory capacity (P＜0.05). U87 SLCs could form larger subcutaneous tumors with fewer inoculated cells (P＜0.05), and grew faster in vivo with stronger tumorigenic ability. Conclusions U87 SLCs have typical stemness characteristics and may function as tumor cell model with higher stemness properties.

miR-142-3p inhibits cerulein-induced apoptosis of rat pancreatic exocrine cell line AR42J by regulating Hmgb1

SU Shixiang, WANG Yuyang, QIN Zongshuai, HUANG Guixiang, XU Jian, CEN Lanying, QIN Yueqiu

2024, 44(1):  23-30.  doi:10.16352/j.issn.1001-6325.2024.01.0023

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Objective To investigate the effect of miR-142-3p on the apoptosis of rat pancreatic exocrine cell line AR42J by regulating Hmgb1. Methods AR42J cells were divided into blank group (blank), acute pancreatitis model group (AP, 100 nmol/L cerulein for 24 h), and then transfected with miR-142-3p mimics, mimics NC,miR-142-3p inhibitor and inhibitor NC, respectively. The cells in the model group were recorded as miR-142-3p mimics group, mimics NC group, miR-142-3p inhibitor group and inhibitor NC.The expression of miR-142-3p in cells was detected by RT-qPCR. The protein expressions of HMGB1, caspase-3, Bax and Bcl-2 were detected by Western blot. Hoechst staining was used to determine cell apoptosis. The apoptosis rate of cells was detected by flow cytometry. The targeting relationship between miR-142-3p and Hmgb1 was determined by dual luciferase reporter gene assay. Results Compared with blank control group, the expression level of miR-142-3p in the AP group was significantly down-regulated (P＜ 0.01), the expression level of HMGB1 and caspase-3 proteins was up-regulated(P＜ 0.05), the expression level of Bax protein was significantly up-regulated (P＜ 0.01), the expression level of Bcl-2 protein was significantly decreased(P＜ 0.01) and the apoptosis rate increased significantly (P＜ 0.01). Compared with the mimics NC group, the level of miR-142-3p in the miR-142-3p mimics group was significantly up-regulated(P＜ 0.01), the expression of HMGB, caspase-3 and Bax proteins was significantly down-regulated (P＜ 0.01), the expression of Bcl-2 protein was up-regulated (P＜ 0.05), and the apoptosis rate decreased significantly (P＜ 0.01).Compared with inhibitor NC group, the expression level of miR-142-3p in miR-142-3p inhibitor group was down-regulated (P＜ 0.05), the expression levels of HMGB1, caspase-3 and Bax proteins were significantly up-regulated (P＜ 0.01), the expression level of Bcl-2 protein was decreased (P＜ 0.05) and the apoptosis rate increased significantly (P＜ 0.01) .The dual luciferase reporter gene assay showed that Hmgb1 was the target gene of miR-142-3p. Conclusions 1)The expression of miR-142-3p was low in the model group. 2)miR-142-3p can inhibit the apoptosis of AR42J cells by inhibiting the expression of Hmgb1.

Rapamycin attenuates ioversol-induced acute kidney injury in rat models

LI Qingju, YU Ran, CHEN Jiajia, CHEN Haoyu, SONG Jian, WANG Wanpeng

2024, 44(1):  31-36.  doi:10.16352/j.issn.1001-6325.2024.01.0031

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Objective To investigate the role of autophagy in contrast-induced acute kidney injury (CI-AKI) in rats and to explore its possible mechanism. Methods SD rats were randomly divided into control group, acute kidney injury model group (intravenous injection of contrast medium ioversol via tail vein; model), rapamycin(RAPA) group and hydroxychloroquine (HCQ)group. Blood urea nitrogen(BUN) and serum creatinine(Scr) contents were measured and the potential change foun in renal pathology was detected by HE staining and microscopy. Transmission electron microscopy was used to observe auto-phagy-related changes in ultrastructure.Western blot was used to observe the expression of microtubule-associated protein 1 light chain 3(LC3)Ⅱ/LC3Ⅰ,ubiquitin-binding protein p62 and Histone deacetylase 4(HDAC4). The expression of HDAC4 was also observed by RT-qPCR. Results Compared with control group, the level of BUN, Scr and HDAC4 expression in the model and HCQ group was increased (P＜0.01),the proximal tubules of the kidney were significantly damaged. In the model group, autophagososomes and autolysosomes increased, accompanied by an increase of LC3Ⅱ/LC3Ⅰ and a decrease in the p62 level (P＜0.05,P＜0.01); Compared with model group, there were more autophagosomes and autolysosomes were found in RAPA group(P＜0.01), accompanied by increased LC3Ⅱ/LC3Ⅰ ratio and decrease in the p62 and HDAC4(P＜0.05,P＜0.01). In contrast, the number of autophagy related structures decreased in HCQ group(P＜0.01), accompanied by the simultaneous increase of LC3Ⅱ/LC3Ⅰ, p62 and the increase of HDAC4(P＜0.01). Conclusions Ioversol may induce autophagy activation, while enhancing autophagy by RAPA alleviates CI-AKI induced renal dysfunction. The mechanism is potentially atributed to the regulation of HDAC4.

Molecular mechanism of ATF6 regulating the reproduction related gene HSPA1L

WANG Yuanyuan, ZHU Xilin, WU Xiaopan, LIU Ying

2024, 44(1):  37-42.  doi:10.16352/j.issn.1001-6325.2024.01.0037

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Objective To explore the effect of endoplasmic reticulum stress activating transcription factor 6 (ATF6) on the expression of reproduction related gene heat shock protein family A member 1 like (HSPA1L) and preliminarily clarify its regulatory molecular mechanism. Methods The ATF6 over-expression plasmid was transfected into HEK-293T cells and the over-expression efficiency was verified by RT-qPCR and Western blot. The transcriptome sequencing information of testis tissue of male ATF6 knockout mice was used to screen five reproduction related genes downstream of ATF6. The dual luciferase reporter assay selected the downstream genes with high promoter activity and detected the effect of over-expression of ATF6 on the promoter activity of downstream genes.The possible binding sites of ATF6 and downstream gene promoters were predicted by gene-regulation. RT-qPCR and Western blot were used to detect the effect of over-expression of ATF6 on downstream gene expression in HEK-293T cells. Whether ATF6 binds to downstream gene promoters was determined by electrophoretic mobility shift assay (EMSA). Results The expression of ATF6 mRNA (P＜0.001) and protein (P＜0.001 and P＜0.05) in HEK-293T cells was significantly increased after transfection. HSPA1L(P＜0.001 and P＜0.05), a reproductive related gene downstream of ATF6 was screened by transcriptome sequencing and dual luciferase reporter assay. ATF6 promoted the truncated promoter activity of HSPA1L(P＜0.001). After over-expression of ATF6, the expression of HSPA1L was significantly increased(P＜0.001). The differences were statistically significant. ATF6 protein could bind to the aagtcgtcac DNA sequence of HSPA1L promoter. Conclusions ATF6, a key molecule of endoplasmic reticulum stress, regulates the expression of reproduction related gene HSPA1L by binding to the promoter of HSPA1L.This result will lay a foundation for further research on the prevention or treatment of endoplasmic reticulum stress (ERS) related male infertility.

Effects of lncRNA FEZF1-AS1 on proliferation, migration and invasion through regulating EZH2 of lung interstitial cells

WANG Chunyan, WANG Ping, SONG Longfei, LIU Yongquan, MAN Jun

2024, 44(1):  43-50.  doi:10.16352/j.issn.1001-6325.2024.01.0043

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Objective To investigate the effects of long non-coding RNA FEZ family zinc finger 1 antisense RNA 1(lncRNA FEZF1-AS1) on enhancer of zeste homolog 2(EZH2) in regulation of proliferation, migration, invasion and epithelial-mesenchymal transition (EMT) of pulmonary interstitial cells and its mechanism. Methods The A549 cells human lung adenocarcinoma cell line were divided into control group and model group [model cells were induced into lung interstitial cells after being treated with transforming growth factor β1(TGF-β1)20 ng/mL for 48 h]. The protein expression of E-cadherin, N-cadherin and vimentin in each group was detected by Western blot. The expression of lncRNA FEZF1-AS1 and EZH2 in the two groups was detected by RT-qPCR. Cells in the transfection group were divided into si NC group,lncRNA FEZF1-AS1+OE vector group and si lncRNA FEZF1-AS1+OE EZH2 group. Cell proliferation was examined by CCK-8 method, cell migration was detected by cell scratch, and cell invasion was detected by Transwell assays. The protein expression of E-cadherin, N-cadherin, vimentin and EZH2 in each group was detected by Western blot. The direct binding effect of FEZF1-AS1 and EZH2 was determined by RNA immuno-precipitation (RIP). Results Compared with the control group, the protein expression level of E-cadherin in the model group was significantly decreased (P＜0.05),and the protein expression of N-cadherin and vimentin was significantly increased(P＜0.05). Compared with the control group, the expression level of lncRNA FEZF1-AS1 and EZH2 genes was significantly increased in the model group (P＜0.05). Compared with si NC group, the proliferation, migration and invasion ability of si lncRNA FEZF1-AS1+OE vector group were decreased, the expression of E-cadherin protein was increased while the expression of N-cadherin, vimentin and EZH2 was decreased(P＜0.05). Compared with si lncRNA FEZF1-AS1+OE vector group, the proliferation, invasion and migration of si lncRNA FEZF1-AS1+OE EZH2 group were increased (P＜0.05). E-cadherin expression was decreased, while N-cadherin, vimentin and EZH2 expressions were increased (P＜0.05). RIP experiment further confirmed that lncRNA FEZF1-AS1 had direct binding effect with EZH2. Conclusions LncRNA FEZF1-AS1 can promote the proliferation, invasion, metastasis and EMT process of pulmonary fibrosis cells by regulating EZH2.

Marein alleviates myocardial fibrosis in diabetic mice

TIAN Yating, ZHANG Fang, ZHANG Boxiang, LI Tian

2024, 44(1):  51-56.  doi:10.16352/j.issn.1001-6325.2024.01.0051

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Objective To study the effect of marein on myocardial fibrosis in diabetic mice. Methods Ten leptin receptor gene defective heterozygous (db/m) mice aged 5-6 weeks were selected as the control group and 30 diabetic mice with leptin receptor gene defective db/db were divided into: db/db group (db/db,n=10), metformin (Met) positive group (280 mg/kg daily,n=10) and marein drug intervention group (50 mg/kg,n=10). After continuous administration for 8 weeks, the cardiac morphological changes were observed by HE staining and Masson staining. The distribution and expression of vimentin were detected by immunohistochemistry method. The expression of fibronectin, vimentin, and transforming growth factor-β1(TGF-β1)protein in cardiac tissue was detected by Western blot. Results Myocardial fiber hypertrophy was observed in db/db group, and myocardial structural damage was improved in metformin group and marein group. Compared with db/m group, the expression of myocardial collagen fiber in db/db group increased (P＜0.01), while the expression of myocardial collagen fiber in metformin group and marein group decreased (P＜0.01). Compared with the control group, the expression of vimentin in myocardial tissue of db/db group was significantly increased(P＜0.01), while the expression of vimentin in metformin group and marein group was significantly decreased (P＜0.01). The expression of fibronectin, vimentin and TGF-β1 in db/db group was significantly increased as compared with those in db/m group (P＜0.01), while the expression of fibronectin, vimentin and TGF-β1 in metformin group and marein group were significantly decreased (P＜0.01). Conclusions Marein improves myocardial fibrosis in diabetic db/db mice.

Increased expression of m⁶A demethylase FTO inhibits the proliferation of nasopharyngeal carcinoma cells

LIAO Zhencheng, YANG Siyi, WU Ping'an

2024, 44(1):  57-62.  doi:10.16352/j.issn.1001-6325.2024.01.0057

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Objective To investigate the expression of N6 methyladenine (m6A) demethylase human fat mass and obesity-associated(FTO) protein in nasopharyngeal carcinoma (NPC), and the effect of over-expression of FTO on the proliferation of nasopharyngeal carcinoma in vitro and in vivo. Methods Immunohistochemistry method was used to detect the expression of FTO protein in nasopharyngeal carcinoma tissues and para-cancerous tissues; The dominant expression cell line of FTO was screened, the over-expression FTO cell line was constructed. The cell proliferation was examined by soft-agar method. A mouse tumor model was developed for measurement of tumor growth. Results The expression of FTO in nasopharyngeal carcinoma tissues was lower than that in adjacent tissues. Low expression of FTO promoted proliferation of NPC cells, while over-expression of FTO reversed this effect. Conclusions FTO inhibits proliferation of nasopharyngeal carcinoma and this result may provide an experimental technology in searching therapeutic targets of chemotherapy for nasopharyngeal carcinoma.

Construction and drug resistance evaluation of cell strains which stably express FIP1L1-PDGFRA protein and its mutants

LIU Jingwen, SONG Haoxin, ZHU Lei

2024, 44(1):  63-68.  doi:10.16352/j.issn.1001-6325.2024.01.0063

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Objective To construct mouse BaF3-FIP1L1-PDGFRA (F/P), BaF3-F/P-T674I and BAF3-F/P-D842V pre-B cell strains which stably express F/P fusion protein and its T674I and D842V mutants in order to evaluate their activity by checking their responses to tyrosine kinase inhibitors (TKIs). Methods Lentivirus infected technique was used to transfect the target gene into BaF3 cells. RT-qPCR was used to detect mRNA expression, and CCK-8 method was used to detect the inhibitory effect of TKIs on the proliferation of stable cell strains. Results The constructed BaF3-F/P, BaF3-F/P-T674I and BAF3-F/P-D842V cell strains all transcripted FIP1L1 and PDGFRA mRNA. They exhibited malignant phenotypic characteristics of proliferation independent of IL-3 and sensitivity to corresponding TKIs. Conclusions The pre-B-cell strains stably expressing F/P and its T674I and D842V mutants are successfully constructed, which provide a good cell model for the development of compounds targeting at those molecules.

Sodium hydrosulfide increases glutathione synthesis in cardiac muscle cell line HL-1 under hyperglycemia and hyperlipidemia

ZHANG Weicai, LIU Siren, WANG Shangnong

2024, 44(1):  69-76.  doi:10.16352/j.issn.1001-6325.2024.01.0069

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Objective To investigate sodium hydrogen sulfide(NaHS) with function of regulating glutathione(GSH) synthesis to reduce reactive oxygen species(ROS) production in type 2 diabetic cardiomyopathy (DCM). Methods Mouse cardiomyocyte cell line HL-1 was incubated with high concentration of glucose (HG:40 mmol/L) and palmitate (Pal:500 μmol/L) as a cell model of type 2 DCM. HL-1 cells were incubated with NaHS (100 μmol/L), DL-propargylglycin(PPG,1 mmol/L) and N-acetyl-l-cysteine(NAC,5 mmol/L), respectively for 72 hours. The expression of cystathionine-γ-lyase(CSE) and the key enzymes of glutathione production was tested by Western blot. Dihydroethidium(DHE) and dichlorofluoromethane(DCFH) were used to detect the content of ROS in HL-1 cells. Cell viability was detected by CCK8 kit. The content of total GSH was detected. The interaction between muscle specific ring finger protein 1(Murf1) and nuclear factor erythroid-derived 2-related factor 2(Nrf2) and Nrf2 ubiquitylation was determined by co-immunoprecipitation(co-IP). Results Compared with control group, the expression level of CSE,solute carrier family 7 members 11(SLC7A11), glutamate cysteine ligase C(GCLC), glutamate cysteine ligase M(GCLM) and glutathione synthetase(GSS) in HL-1 cells treated incubated with high glucose and palmitate was decreased, however, NaHS was found to restore it. NaHS reduced the content of ROS in HL-1 cells treated with high glucose and palmitate. The interaction between murf1 and Nrf2 was confirmed by co-immunoprecipitation(Co-IP). Compared with NaHS group, the ubiquitylation level of Nrf2 was enhanced in high glucose and palmitate group. Conclusions Sodium hydrosulfide may reduce the ubiquitylation level of Nrf2 and promote the expression of key enzymes of GSH synthesis.

Wogonin promotes Th17/Treg cell balance in rat models with autoimmune hepatitis

DENG Juan, WANG Xiufang, SUN Ruiqing

2024, 44(1):  77-83.  doi:10.16352/j.issn.1001-6325.2024.01.0077

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Objective To investigate the impacts of wogonin (WG) on Th17/Treg cell balance in autoimmune hepatitis (AIH) rats. Methods A total of 10 rats were randomly selected as the control group. The remaining rats were injected with concanavalin A (ConA, 12.5 mg/kg) solution via tail vein to construct AIH model rat, which were randomly divided into AIH group, L-WG group (10 mg/kg), M-WG group (20 mg/kg), H-WG group (30 mg/kg), H-WG+VPA group (30 mg/kg WG+300 mg/kg Notch signal pathway activator VPA), 10 rats in each group and administered once a day for 10 days. Serum inflammatory factors and liver function indexes were detected by ELISA; HE staining was used to observe the pathological morphology of liver tissue; the level of spleen Th17/Treg cells was detected by flow cytometry; Western blot was used to detect the expression of spleen retinoid acid related orphan receptor γ t (RORγt), fork head box protein P3 (Foxp3) and liver Notch signal pathway proteins. Results The liver tissue structure of control group was normal and the staining was clear; In AIH group, the cells of liver tissue showed edema, the increase of cell volume led to the compression and narrowing of liver sinuses, and a large number of inflammatory cell infiltration and a small amount of necrosis occurred. The contents of alanine aminotransferase (ALT), aspartate aminotransferase (AST), interleukin (IL)-17 and IL-23, level of Th17 cells, Th17/Treg, the expression of RORγt, Notch, hes family bHLH transcription factor 1 gene (HES1) and hes related family bHLH transcription factor with YRPW motif 1 (HEY1) protein in AIH group were greatly higher than those in control group (P＜0.05), the contents of IL-10 and TNF-β, level of Treg cells, and level of Foxp3 protein were greatly lower(P＜0.05); Compared with AIH group, the liver injury in L-WG group, M-WG group and H-WG group was improved, the contents of ALT, AST, IL-17 and IL-23, level of Th17 cells, Th17/Treg, the expression of RORγt, Notch, HES1 and HEY1 protein were greatly lower (P＜0.05), the contents of IL-10 and TNF-β, level of Treg cells, and level of Foxp3 protein were greatly higher (P＜0.05); VPA reversed the improvement effect of H-WG on AIH rats. Conclusions WG could promote Th17/Treg cell balance in AIH rats by down-regulating Notch signal pathway.

Clinical Sciences

Investigation on the risk of recurrent immune thrombocytopenia in children and establishment of a predictive model

SHEN Chentao, XIA Yalin, SHENG Yeping, CHU Peipei, LI Jianqin

2024, 44(1):  84-91.  doi:10.16352/j.issn.1001-6325.2024.01.0084

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Objective To investigate the recurrence of immune thrombocytopenia (ITP) in children and to establish a predictive model. Methods A total of 288 children with ITP admitted to Children's Hospital of Wujiang District and Children's Hospital Affiliated to Suzhou University from January 2018 to April 2022 were collected. The factors potentially related to the recurrence of ITP in children were screened . The children in the model group were divided into 2 groups according to the presence or absence of recurrence and the indicators of the 2 groups were compared. After screening the potential influencing factors by LASSO regression and the independent influencing factors of relapse in children with ITP patients by Logstic regression analysis, we constructed a column-line graph model by using R language and validated it. Results A total of 37 (18.47%) of 201 patients in the model group experienced relapse. The age, blood type, duration of disease before treatment, antecedent infections, bleeding, initial treatment regimen, antinuclear antibody titer, initial count and mean platelet volume, initial platelet distribution width, initial peripheral blood lymphocyte count and time length to effective platelet count after treatment were found in the recurrence group versus the non-recurrence group The difference was statistically significant(P＜0.05).The results of multifactorial logistic regression analysis performed on the basis of LASSO regression showed that blood type, duration of illness before treatment, antecedent infection, initial treatment regimen, initial peripheral blood lymphocyte count, and time to effective platelet count after treatment were independent influences on the conversion of cough variant asthma to classic asthma in children. Based on the results of the multifactorial analysis, a column chart model for predicting ITP recurrence in children was developed in R. The results of the receiver operating characteristic(ROC) analysis showed that the area under curve(AUC) of the column chart model for predicting ITP recurrence in children in the modeling group was 0.867[95% CI(0.796, 0.938)] with a sensitivity of 84.2% and a specificity of 73.1%, and that in the validation group, the AUC was 0.838 [95% CI(0.765), 0.911] with a sensitivity of 82.3% and a specificity of 78.4%, 0.911)] sensitivity was 82.3% and specificity was 78.4%. The Bootstrap method was used to repeat the sampling 1 000 times, and the validation group was used for validation. The results of the calibration curve showed that the prediction curves of the model group and the validation group were basically fitted with the standard curve, suggesting that the model prediction accuracy was high. The results of the decision curve analysis of the model group showed that the net benefit rate of patients was greater than zero when the probability threshold of the column line graph model of predicting ITP recurrence in children was 0.15-0.75. Conclusions ITP recurrence in children is mainly affected by the patient's age, blood type, and pre-treatment course of the disease, and the column-line diagram model based on these factors has a high accuracy and differentiation for ITP recurrence in parenting children.

Factors affecting nosocomial death in elderly patients with COVID-19 and construction of a risk predictive model

DAI Jingrong, XIAO Bao, LI Lin, HU Jiangying, LIU Bin

2024, 44(1):  92-97.  doi:10.16352/j.issn.1001-6325.2024.01.0092

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Objective To study the factors affecting hospital death in elderly patients with novel coronavirus infection/disease 2019(COVID-19), and to build a risk prediction model. Methods According to the diagnostic criteria of Diagnosis and Treatment Protocol for COVID-19 Infection (Trial 10th Edition). Totally 775 elderly patients (≥60 years old) diagnosed as COVID-19 infection in the emergency department and fever clinic of the First Hospital of Changsha were selected as the research objects. General data and serum biomarkers of patients were collected. After treatment, the patients' data were divided into survival group and hospital death group. Binary Logistic regression was used to screen the independent influencing factors of death, and ROC curve was used to analyze the predictive value of related indicators on hospital death. Results After treatment, 712 patients (91.9%) survived and 63 patients (8.3%) died in hospital. Binary Logistic regression analysis showed that: ≥90 years old [OR=5.065, 95% CI (1.427,17.974)], type 2 diabetes mellitus [OR=3.757, 95% CI (1.649,8.559)], COPD[OR=5.625,95% CI (2.357,13.421)], monocyte ratio [OR=0.908,95% CI (0.857,0.963)], plasma fibringen [OR=1.376,95% CI (1.053,1.800)] and lactate dehydrogenase [OR=1.005,95% CI (1.001,o1.008)] were independent factors of in-hospital death (P＜0.05). The predictive value of diabetes mellitus + COPD + age + monocyte ratio + plasma fibrinogen + lactate dehydrogenase was proved in hospital death from COVID-19 infected patients: the area under the curve(AUC) was 0.883 (95% CI: 0.827,0.940,P＜ 0.001), the critical value ≥0.710 suggested the risk of death in hospital, the specificity was 0.851, the sensitivity was 0.857. Conclusions The hospital mortality of the elderly after COVID-19 infection is higher and closely related to type 2 diabetes, COPD, monocyte ratio, plasma fibrinogen and lactate dehydrogenase.

Development of a predictive model for perioperative blood transfusion in elderly patients undergoing unilateral total hip arthroplasty

ZANG Han, HU Ai, XU Xuanqi, XU Li

2024, 44(1):  98-102.  doi:10.16352/j.issn.1001-6325.2024.01.0098

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Objective To analyze risk factors for perioperative blood transfusion in elderly patients undergoing unilateral primary total hip arthroplasty and develop a prediction model. Methods The study retrospectively collected 467 elderly patients receiving unilateral primary total hip arthroplasty between January 2013 and October 2021 at Peking Union Medical College Hospital. The 70% of the data were used as the training set and the 30% of the data were used as the testing set. Patients were divided into the transfusion and no-transfusion groups based on the presence or absence of perioperative blood transfusion. Univariate analysis and multivariable logistic regression were conducted to analyze patient demographic characteristics, surgical information, and preoperative laboratory tests for identifying risk factors. Clinical experience was combined to establish a prediction model and draw the nomogram. The receiver operating characteristic(ROC) curve and calibration curve were used to evaluate the model in the testing set. Results A total of 91 patients(19.5%) received perioperative blood transfusion. Multivariable logistic regression suggested the history of coronary artery disease, prolonged operation time, and lower preoperative hemoglobin were risk factors for perioperative blood transfusion(P＜0.05). The prediction model was constructed based on the results of statistical analysis and clinical experience, including the history of coronary artery disease, operation time, preoperative hemoglobin, age, and American Society of Anesthesiologists(ASA) physical status＞Ⅱ. The area under the receiver operating characteristic curve(AUC) of the model was 0.809. Conclusions The prediction model for perioperative blood transfusion in elderly patients undergoing unilateral total hip arthroplasty had a good performance and could assist in clinical practice.

Mini Reviews

Research progress on the role of lipoprotein- associated phospholipase A2 in acute coronary syndrome

GUO Jianhong, WANG Yaling, CUI Wenguang

2024, 44(1):  103-107.  doi:10.16352/j.issn.1001-6325.2024.01.0103

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Lipoprotein-associated phospholipase A2(Lp-PLA2) is a protein composed of 441 amino acids, which can promote the aggregation of inflammatory cells to the inflammatory response site and the release of inflammatory factors. It can promote the synthesis of matrix metalloproteinases, increase the number of foam cells and extra cellular matrix in atherosclerotic plaque, attenuate plaque fiber cap and prone to rupture, thus promote the onset of acute coronary syndrome(ACS). Therefore, the determination and regulation of Lp-PLA2 levels in patients with ACS are clinically significant.

Progress of researches on inducing hepatic stellate cell ferroptosis for treatment of liver fibrosis

SUN Lijie, HAO Dandan

2024, 44(1):  108-113.  doi:10.16352/j.issn.1001-6325.2024.01.0108

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Emerging evidence suggests that ferroptosis, an iron-dependent form of regulated cell death, plays a critical role in the genesis of liver fibrosis through inducing hepatic stellate cell (HSC) ferroptosis to inhibit liver fibrosis or inducing hepatic ferroptosis to potentiates liver fibrosis. Pharmacologically targeting at ferroptosis with its inducers can slow down the progression of liver fibrosis in vitro and in vivo model. This review suggests that pharmacological induction of HSC ferroptosis might be used as a potential novel targeted therapy for the treatment of liver fibrosis.

Research progress of hypoxia inducible factor 1α in the mechanism of diabetic kidney disease

ZHANG Yahui, BAI Qiong

2024, 44(1):  114-118.  doi:10.16352/j.issn.1001-6325.2024.01.0114

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Hypoxia-inducible factor-1α (HIF-1α) is a nuclear transcription factor. Under high glucose and hypoxia conditions, the expression of HIF-1α is elevated, result in the increased expression of its downstream target genes VEGF, HO-1 and BNIP3, which affect angiogenesis, extra cellular matrix deposition, iron metabolism, and mitophagy, participating in the occurrence and development of diabetic kidney disease (DKD). In addition, HIF-1α promotes inflammation and renal fibrosis by affecting the production of cytokines in DKD.

Progress of the effect of hydroxyacyl- coenzyme A dehydrogenase in cancer development and its mechanism

WU Guojia, ZHAI Shujie, SUN Xiao, HUANG Yiran, LI Yongmei, SUN Li

2024, 44(1):  119-123.  doi:10.16352/j.issn.1001-6325.2024.01.0119

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A close relationship between fatty acid metabolism and cancer development is well-established. The hydroxyacyl-coenzyme a dehydrogenase (HADH), a key enzyme in fatty acid beta-oxidation, has recently been identified as an anti-oncogenic factor in various cancers and an oncogenic factor in conditions like acute myeloid leukemia. In cancer cells, HADH not only directly catalyzes fatty acid beta-oxidation but also indirectly influences multiple signaling pathways such as PPAR, TNF-α, JAK-STAT3, PI3K/Akt, IFN-γ, MAPK, and non-canonical Wnt signaling pathways, affecting cancer cell proliferation and migration. HADH shows promise as a potential tumor biomarker for diagnosis, treatment, and prognosis in different cancer types, holding significant clinical value.

Research progress of artificial blood

REN Feng, JIN Yishan, CHEN Lu, RONG Long, YU Chunhong

2024, 44(1):  124-130.  doi:10.16352/j.issn.1001-6325.2024.01.0124

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Artificial blood is a type of liquid preparation with oxygen-loading capacity and can temporarily substitute some function of blood. The developed artificial blood can be divided into four categories: artificial synthetic hemoglobin, artificial red blood cells made from natural hemoglobin, perfluorocarbons, and stem cell-differentiated red blood cells. This review focuses on the domestic and foreign research progress of artificial blood in recent years, and discusses its clinical application value, development trend, and future research, in order to provide new ideas to the development the artificial blood products and promote clinical application.

Research advances on optical characteristics in central serous chorioretinopathy

XIAO Xingyu, DAI Rongping

2024, 44(1):  131-135.  doi:10.16352/j.issn.1001-6325.2024.01.0131

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Central serous chorioretinopathy (CSC) is a chorioretinal disease that causes idiopathic serous retinal detachment (SRD), which is associated with one or more areas of pigment epithelial detachment (PED) or defect in the retinal pigment epithelium, also with characteristic ocular structural changes. CSC was classified as pachychoroid spectrum diseases (PSD); recent studies have found it mainly in Haller layer. Recent studies focused on the thick sclera in CSC patients, illustrated the close relation between which and choroidal circulation and put forward the probable pathogenesis similar to uveal effusion syndrome (UES). In addition, short axial length, hyperopia and shallow anterior chamber are also the characteristics in CSC patients, indicating that CSC is the disease not limiting to posterior oculus, but involving the whole oculus. This review summarizes the latest research advances on optical characteristics in CSC, providing the new ideas for further research on pathogenesis of CSC.

Medical Education

Teaching reform of Traditional Chinese Medicine(TCM) acupuncture course in Nursing College of Peking Union Medical College

CHEN Suhui, XU Hong, ZHANG Yamin, SUN Hua, LI Ying, WU Qunli

2024, 44(1):  136-140.  doi:10.16352/j.issn.1001-6325.2024.01.0136

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Objective To reform the teaching content of acupuncture and moxibustion for undergraduate of Nursing College and to evaluate the effectiveness of the teaching reform, so to provide constructive information for optimizing the curriculum. Methods To revise the teaching content of Traditional Chinese Medicine(TCM) acupuncture course based on the results of previous questionnaires, including reducing boring theoretical knowledge, increasing commonly used acupuncture techniques and clinical case analysis, organizing all undergraduates attending the TCM teaching in the Nursing College of Peking Union Medical College to fill out the “Acupuncture Course Content Questionnaire”in 2023. The content covers four aspects: students' basic cognition of acupuncture and moxibustion before the course, students' learning feedback,students'learning effectiveness after the course and students' suggestions for further optimization of acupuncture and moxibustion course. Results Totally 149 students participated in the questionnaire survey and 73.15% of them were interested in the acupuncture course. There were 62.42% of the students thought that the most difficult part of the course was the “Acupuncture Points”, and 71.81% of them complained the most constraining factor to the learning effectiveness was the difficulty of memorizing the meridians and acupoints. The most interesting part of the course was “Overview of Acupuncture Treatment and Acupuncture Treatment of Common Diseases”, which accounted for 44.3% of the students. Through the study, 88.59% of the students were willing to recommend acupuncture treatment to patients with indications.Students' suggestions for improvement of the acupuncture course was increase of classroom practice or learning by observation of operation videos. Conclusions The reformed acupuncture course highlights the practicability, improves the learning interest and subjective initiative, but the training of practice skill still needs to be strengthened.