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15 February 2022, Volume 50 Issue 2 Previous Issue    Next Issue

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MiR-1307-3p promotes the proliferation and migration of breast cancer cells by targeting ISM1 #br# LU Xuyang, LIU Yi, LI Xiaohui, HUANG Xin, HUANG Cuixia, MA Shuxiang, DOU Weiyu, ZENG Qiyan 2022, 50 (2):  113-119.  doi: 10.11958/20210892 Abstract ( 512 )   PDF (2102KB) ( 2582 )   Objective To explore the possible molecular mechanism of miR-1307-3p in promoting proliferation and migration by targeting ISM1 in breast cancer cells. Methods (1) Cancer Genome Atlas (TCGA) database was used to analyze the expression level of miR-1307-3p and its relationship with clinical indicators in breast cancer patients. (2) qPCR, CCK-8 assay, Scratch test and flow cytometry were used to detect the effects of miR-1307-3p over-expression or silence on the expression of miR-1307-3p, cell proliferation, migration and apoptosis in breast cancer MCF-7 cells, respectively. (3) The target gene of mir-1307-3p was predicted by bioinformatics analysis, and verified by double luciferase reporter gene experiment. Results The relative expression levels of miR-1307-3p were significantly up-regulated both in breast cancer cell lines and breast cancer tissues. Overexpression of miR-1307-3p significantly promoted cell proliferation and migration in MCF-7 cells; on the contrary, miR-1307-3p inhibitor significantly inhibited cell migration and induced cell apoptosis. ISM1 may be the target genes of miR-1307-3p. The expression level of ISM1 was significantly lower in breast cancer tissues than that in normal breast tissues, and was significantly correlated with clinical stages. Conclusion MiR- 1307-3p can promote the proliferation and migration of breast cancer cells, which may affect the occurrence and development of breast cancer by targeting ISM1 gene. Related Articles | Metrics

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Mechanism of PTEN/mTOR pathway in high glucose-induced apoptosis of human villous trophoblasts HTR-8/SVneo #br# PENG Qing, WEN Yanjing△, LI Qian, LI Man, CHANG Meiying 2022, 50 (2):  120-124.  doi: 10.11958/20211542 Abstract ( 468 )   PDF (604KB) ( 2556 )   Objective To study the mechanism of high glucose-induced apoptosis of human villous trophoblasts HTR- 8/SVneo. Methods Human villous trophoblasts HTR-8/SVneo were cultured in vitro and divided into the blank control group, the high glucose group, the negative control group (NC), the phosphatase and tensin homology deleted on chromosome ten (PTEN) -siRNA group (inhibition of PTEN expression) and the mammalian target of rapamycin (mTOR) pathway inhibitor group (GDC-0349 group). Methyl thiazolyl tetrazolium (MTT) assay and flow cytometry were used to detect the proliferation of HTR-8/SVneo cells and the apoptosis of HTR-8/SVneo cells, respectively. The expression of apoptosis related proteins and proteins related to PTEN/mTOR pathway were detected by Western blot assay. Results Compared with the blank control group, HTR-8/SVneo cell proliferation inhibition rate, apoptosis rate and Bcl-2 related X protein (Bax), cleaved caspase-3 and PTEN protein expression levels were significantly increased in the high glucose group (P＜0.05), while the expression levels of B-cell lymphoma-2 (Bcl-2), phosphorylation phosphatidylinositol-3-kinase (p-PI3K)/PI3K, phosphorylation protein kinase B (p-AKT)/AKT and phosphorylation mTOR (p-mTOR)/mTOR protein were significantly decreased (P＜0.05). Compared with the high glucose group and the NC group, the HTR-8/SVneo cell proliferation inhibition rate, apoptosis rate, Bax, cleaved caspase-3 and PTEN protein expression levels were significantly decreased in the PTEN-siRNA group (P＜0.05), while the expression levels of Bcl-2, p-PI3K/PI3K, p-AKT/AKT and p-mTOR/mTOR protein were significantly increased (P＜0.05). Compared with the PTEN-siRNA group, the proliferation inhibition rate, apoptosis rate and the protein expression levels of Bax, cleaved caspase-3 were significantly increased in the GDC-0349 group (P＜0.05), while the expression levels of Bcl-2, p-PI3K/PI3K, p-AKT/AKT and p-mTOR/mTOR protein were significantly decreased (P＜0.05). Conclusion High glucose can inhibit the growth and induce apoptosis of human villous trophoblasts HTR-8/SVneo, which may be achieved by up-regulating the expression of PTEN and inhibiting the activation of PI3K/AKT/mTOR pathway. Related Articles | Metrics

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Study on the mechanism of quercetin improving gemcitabine resistance in NSCLC through the EMT pathway mediated by miR-101/EZH2 axis QIAN Qiyu, YUAN Gaili, MA Shanshan, LUO Limei, XU Yu 2022, 50 (2):  125-130.  doi: 10.11958/20211879 Abstract ( 446 )   PDF (502KB) ( 2502 )   Objective To investigate the effect of quercetin on epithelial-mesenchymal transdifferentiation (EMT) of non-small cell lung cancer (NSCLC) and its molecular mechanism in improving gemcitabine (Gb) resistance from the perspective of microRNA-101 (miR-101)/Zeste gene homologous protein 2 (EZH2) axis. Methods A549 and A549/Gb cells were cultured with different concentrations of quercetin, and CCK-8 method was used to screen suitable quercetin concentration for the follow-up test. A549 cells were divided into the blank group, the A549 + EMT inducer group, the A549+ quercetin group and the quercetin + EMT inducer group. A549 cells and A549/Gb were divided into the A549 group, the A549/Gb group, the A549/Gb + quercetin group, the A549/Gb + EMT inducer group, the miR-101-inhibitor group, the quercetin+miR-101-inhibitor group and the miR-NC group. Real-time fluorescence quantitative PCR (qPCR) was used to detect the expression level of miR-101 in each group. The expression of EZH2, transforming growth factor (TGF) - β1/ drosophila, mother DPP homolog 4 (SMAD4), p-SMAD4, and EMT marker proteins E-cadherin, N-cadherin and Vimentin were detected by Western blot assay. The optical density (OD) of each group was measured by CCK-8 method, and IC50 and IR were calculated. Results The IC 50 of quercetin for A549 and A549/Gb were 64 and 128 μmol/L, respectively. After EMT inducer enhanced the EMT characteristics of A549 cells (the expression levels of N-cadherin and Vimentin were increased, the expression of E-cadherin was decreased), the expression of miR-101 was decreased, EZH2, TGF-β1 and pSMAD4/SMAD4 protein expression, IC50 and IR increased (P＜0.05). Quercetin could inhibit the EMT characteristics of A549 cells, reduce IC50 and IR, up-regulate miR-101, inhibit EZH2, TGF-β1 and p-SMAD4/SMAD4 protein expression (P＜0.05). EMT inducers could reverse the above effects of quercetin (P＜0.05). Compared with A549 cells, the EMT characteristics increased in A549/Gb cells, and IC50, IR, EZH2, TGF- β1 and p-SMAD4/SMAD4 expressions were all increased (P＜0.05). The inhibition of miR-101 or treatment with EMT inducer could further increase the EMT characteristics, IC50, IR, EZH2, TGF-β1 and p-SMAD4/SMAD4 protein expression of A549/Gb cells (P＜0.05). Quercetin could inhibit the EMT process of A549/Gb cells and reduce its IC50, IR and EZH2, TGF-β1 and p-SMAD4/SMAD4 protein expression (P＜0.05). The inhibition of miR-101 expression could reverse the above effects of quercetin (P＜0.05). Conclusion Quercetin can up-regulate miR-101 and inhibit EZH2 expression, thereby inhibiting EMT process and reducing NSCLC cell drug resistance. Related Articles | Metrics

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Mechanism of LncRNA GAS5 targeting miR-103 to reduce insulin resistance in 3T3L1 adipocytes #br# AO Wen, XU Zaige, BAI Yang, LIU Huishuang 2022, 50 (2):  131-136.  doi: 10.11958/20211644 Abstract ( 484 )   PDF (619KB) ( 2519 )   Objective To explore the mechanism of long non-coding RNA (LncRNA) growth arrest-specific 5 (GAS5) targeting microRNA (miR) -103 to reduce 3T3L1 adipocyte insulin resistance (IR). Methods The 3T3L1 mouse preadipocytes were cultured and induced to differentiation, and oil red O staining was used to identify cell differentiation. The 3T3L1 adipocyte IR model was established. Cells were divided into the control group, the model group, the empty vector group (transfected with pEGFP-C1 empty vector), the GAS5 over-expression group (transfected with pEGFP-C1-GAS5 vector), the GAS5 over-expression + mimic NC group (transfected with pEGFP-C1-GAS5+mimic NC) and the GAS5 overexpression+miR-103 mimic group (transfected with pEGFP-C1-GAS5+miR-103 mimic). Real-time fluorescent quantitative PCR was used to detect the levels of GAS5 mRNA and miR-103 in cells. liquid scintillation method was used to detect glucose uptake capacity. Western blot assay was used to detect the expression levels of insulin receptor substrate-1 (IRS-1), p-IRS-1, peroxisome proliferator-activated receptor γ (PPARγ), glucose transporter 4 (GLUT4), and protein kinase B (AKT) and p-AKT proteins. Dual luciferase was used to identify the targeting sites of miR-103 and GAS5. Results After induction, the cells were round, the cell body enlarged, the cytoplasm was rich and contained a large number of lipid droplets. The oil red staining was obvious, showing‘finger-ring-like’structure. The model was successfully constructed. Compared with the control group, the GAS5 mRNA level, glucose uptake capacity, p-IRS-1/IRS-1, PPARγ, GLUT4 and pAKT/AKT protein levels decreased in the model group, the empty vector group and the GAS5 over-expression+miR-103 mimic group (P＜0.05), the miR-103 mRNA level in cells increased (P＜0.05). Compared with the model group and the empty vector group, the GAS5 mRNA level, glucose uptake capacity, p-IRS-1/IRS-1, PPARγ, GLUT4 and p-AKT/AKT protein levels increased in the GAS5 over-expression group and the GAS5 over-expression+mimic NC group (P＜0.05), while the miR-103 mRNA level in cells decreased (P＜0.05). Compared with the GAS5 over-expression group and the GAS5 over-expression + mimic NC group, the GAS5 mRNA level, glucose uptake capacity, p-IRS-1/IRS-1, PPARγ, GLUT4 and p-AKT/AKT protein levels decreased in the GAS5 over-expression + miR-103 mimic group (P＜0.05), the miR-103 mRNA level increased (P＜0.05). The complementary binding sites of MiR-103 and GAS5 were verified by the dual luciferase targeting relationship. Conclusion Targeted down-regulation of miR-103 expression after over-expression of GAS5 can reduce IR of 3T3L1 adipocytes. Related Articles | Metrics

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Secretory clusterin attenuated hydrogen peroxide induced cardiomyocyte injury by inhibiting mitophagy #br# HE Jing, SUN Xiaohui, YANG Li, WU Yuliang 2022, 50 (2):  136-142.  doi: 10.11958/20211411 Abstract ( 517 )   PDF (650KB) ( 2591 )   Objective To investigate the effects of secretory clusterin (sCLU) on oxidative-induced cardiomyocyte injury and the underlying mechanism. Methods Rat cardiomyocytes H9C2 were used in this experiment. Experiment 1 was divided into the control group (only medium change) and the H2O2 group (treated with 100 µmol/L H2O2). Experiment 2 was divided into the control group, the pcDNA3.1 group (transfected with pcDNA3.1 empty plasmids), the pcDNA3.1-sCLU group (transfected with pcDNA3.1-sCLU overexpressed plasmids), the H2O2 group and the H2O2+pcDNA3.1 group (transfected with pcDNA3.1 empty plasmids for 48 h, and then treated with 100 µmol/L H2O2). The H2O2+pcDNA3.1-sCLU was transfected with pcDNA3.1-sCLU over-expressed plasmids for 48 h, and then 100 µmol/L H2O2. Experiment 3 was divided into the DMSO group (adding 1% volume of DMSO), the Mdivi-1 group (treated with 10 µmol/L Mdivi-1), the H 2O2+DMSO group (treated with 1% volume of DMSO for 30 min, and then added 100 µmol/L H2O2), the H2O2+Mdivi-1 group (treated with 100 µmol/L Mdivi-1 for 30 min, and then treated with 100 µmol/L H2O2), the H2O2+pcDNA3.1-sCLU group and the H2O2+pcDNA3.1-sCLU+Mdivi-1 group (transfected with pcDNA3.1-sCLU overexpressed plasmids for 48 h, and then treated with 10 µmol/L Mdivi-1 for 30 min, and then added 100 µmol/L H2O2). MTT assay was used to detect cell viability. TUNEL assay was used to detect cell apoptosis. The corresponding Kit was used to detect the activity of reactive oxygen species (ROS), superoxide dismutase (SOD) and malondialdehyde (MDA). The expression of sCLU in cells was detected by qRT-PCR and ELISA. The protein expression levels of CLU, mitochondrial autophagy related proteins PINK1, Parkin and LC3 were detected by Western blot assay. Results In experiment 1, compared with the control group, the relative expression levels of sCLU mRNA, sCLU protein levels in cell culture supernatant and total CLU protein levels in cells were decreased in the H 2O2 group (P＜0.05). In experiment 2, compared with the control group, cell viability, SOD levels and LC3-II/LC3-I ratio were all decreased in the H 2O2 group (P＜0.05). Apoptosis rate, ROS and MDA levels, PINK1 and Parkin protein expression levels were all increased in the H2O2 group (P＜0.05). Compared with H2O2 group, cell viability rate, SOD levels and LC3-Ⅱ/LC3-Ⅰ ratio were increased in H2O2+pcDNA3.1-sCLU group (P＜0.05). Apoptosis rate, ROS and MDA levels, PINK1 and Parkin protein expression levels were all decreased in H2O2+pcDNA3.1-sCLU group (P＜0.05). In experiment 3, compared with DMSO group, mdivi-1 group showed no significant difference in cell viability and apoptosis rate. Cell viability was decreased and apoptosis rate was increased in H2O2+DMSO group (P＜0.05). Compared with the H2O2+DMSO group, cell viability was increased and cell apoptosis rate was decreased in the H2O2+ Mdivi-1 group and H2O2+pcDNA3.1-sCLU group (P＜0.05). There were no significant differences in cell viability and the cell apoptosis rate between the DMSO group and the Mdivi-1 group. Compared with the DMSO group, the cell viability decreased and the cell apoptosis rate increased in the H2O2+DMSO group (P＜0.05). Compared with the H2O2+DMSO group, cell viability increased and cell apoptosis rate decreased in the H2O2+Mdivi-1 group, the H2O2+PCDNA3.1-sCLU group and the H2O2+PCDNA3.1-SCLU+Mdivi-1 group (P＜0.05). There was no significant difference in cell viability between the H 2O2+Mdivi-1 group, H2O2+pcDNA3.1-sCLU group and the H2O2+pcDNA3.1-sCLU+Mdivi-1 group (P＞ 0.05). Conclusion sCLU can protect myocardial cells under oxidative stress, and the mechanism may be related to the inhibition of mitophagy. Related Articles | Metrics

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The effects of exogenous hydrogen sulfide on cognitive function and intestinal barrier function through Nrf2/ARE/HO-1 pathway in schizophrenic rats CHEN Li, MA Yali, LI Benliang 2022, 50 (2):  143-149.  doi: 10.11958/20211303 Abstract ( 377 )   PDF (973KB) ( 2544 )   Objective To investigate the effects of exogenous hydrogen sulfide (H2S) on cognitive function and intestinal barrier function through nuclear factor E2 related factor 2/antioxidant response element/heme oxidase 1 (Nrf2/ARE/ HO-1) pathway in schizophrenic rats. Methods Forty SD rats were randomly divided into the control group, the model group, the H2S intervention group (intraperitoneal injection of 10 mg/kg H2S donor GYY4137), the Nrf2 inhibitor group (intraperitoneal injection of 10 mg/kg Nrf2 inhibitor ATRA) and the H2S+Nrf2 inhibitor group (intraperitoneal injection of GYY4137, ATRA), with eight in each group. Except for the control group, the other groups were injected intraperitoneally with 0.2 mg/kg of MK-801 to construct the schizophrenic rat model. After the model was successfully constructed, intervention was carried out according to the administration method of each group. The model group and the control group were replaced with normal saline, continuous intervention for 7 days. Morris water maze test was used to measure the cognitive function of rats. Hematoxylin-eosin staining (HE) was used to observe the pathological changes of hippocampus and intestine tissue. TUNEL was used to detect the apoptosis of intestinal mucosal cells in rats. Enzyme-linked immunosorbent (ELISA) kit was used to determine the levels of H2S, D-lactic acid, intestinal tissue superoxide dismutase (SOD), MDA, tumor necrosis factor (TNF)-α, and interleukin (IL)-6 in rat plasma. Western blot assay was used to detect the apoptosis of intestinal tissue and Nrf2/ARE/HO-1 pathway related proteins of rats. Results Compared with the control group, the injury of hippocampal tissue and intestinal tissue were aggravated in the model group. The incubation period, plasma D-lactic acid, intestinal mucosal cell apoptosis rate, intestinal Caspase-3, Bax protein, MDA, TNF-α, IL-6, Nrf2 and HO-1 were increased (P＜0.05). The number of puncture, the level of plasma H2S, the height of small intestinal villi, the depth of recess, the level of Bcl-2 protein and SOD decreased in intestinal tissue (P＜0.05). Compared with the model group, the injury of hippocampus and intestine was alleviated in the H2S intervention group. The incubation period, plasma D-lactic acid, intestinal mucosal cell apoptosis rate, intestinal tissue Caspase-3, Bax protein, MDA, TNF-α and IL-6 decreased (P＜ 0.05). The number of puncture, the level of plasma H2S, the height of small intestinal villi, the depth of recess, the levels of Bcl-2, Nrf2, HO-1 protein and SOD in intestinal tissue increased (P＜0.05). Nrf2 inhibitor can partially reverse the beneficial effects of H 2S on cognitive function, intestinal barrier and hippocampus of model rats. Conclusion Exogenous H 2S can significantly improve the cognitive function and intestinal barrier function of schizophrenic rats, which may be achieved by activating the Nrf2/ARE/HO-1 pathway. Related Articles | Metrics

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The expression of six kinds of mitochondrial N-formyl peptides in plasma and pancreatic FPR1 in severe acute pancreatitis rats XIAO Yi, ZHANG Guixian, GAO Ruifang, LI Xia, SHEN Hongsheng, LIU Hongbin 2022, 50 (2):  150-154.  doi: 10.11958/20211715 Abstract ( 447 )   PDF (1185KB) ( 2610 )   Objective To detect the expression levels of six mitochondrial N-formyl peptides (NFPs) in plasma and formyl peptide receptor 1 (FPR1) in pancreas of rats with severe acute pancreatitis (SAP), and to explore the correlation between them and the progression of SAP. Methods Thirty male SD rats were divided into the sham operation group, the SAP model 3 h group and the SAP model 6 h group by random number table method, with 10 rats in each group. In the sham operation group, the pancreas was turned slightly after laparotomy, and then the abdominal cavity was closed. SAP rat model was prepared in the SAP groups by retrograde injection of 5% sodium taurocholate (50 mg/kg) into the biliary duct. The expression levels of six mitochondrial NFPs in plasma were detected by Western Blotting. The pathological changes of pancreatic tissue were observed by hematoxylin-eosin staining. Immunohistochemistry was used to detect the protein expression of FPR1 in pancreas. Results Compared with the sham operation group, pancreatic hemorrhage, acinoid cell necrosis, inflammatory cell infiltration, edema and other pathological changes occurred in the SAP model 3 h group and the SAP model 6 h group, and the pathological scores were significantly increased, especially in the SAP model 6 h group (P＜ 0.05). Immunohistochemical results showed that the expression levels of FPR1 in pancreatic tissue were significantly higher in the SAP model 3 h group and the SAP model 6 h group than that of the sham group, especially in the SAP model 6 h group (P＜0.05). The expression levels of six mitochondrial NFPs were varied. The expression levels of MT-ND1, MT-ND3 and MT-ND6 were gradually increased with the prolonging of SAP modeling time (P＜0.05), while MT-ND4 showed a decreasing trend (P＜0.05). There was no significant difference in the expression of MT-ND5 between the three groups. Conclusion FPR1 and some mitochondrial N-formyl peptides are closely associated with the progression of SAP, and which may be related to the excessive inflammatory response and amplification of inflammatory cascade in SAP. Related Articles | Metrics

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Gastrodin can improve the head twitch behavior of rats with Tourette syndrome DAI Weifeng, HAN Xue, YUE Jiaojiao, GE Guolan 2022, 50 (2):  155-159.  doi: 10.11958/20211208 Abstract ( 592 )   PDF (519KB) ( 2549 )   Objective To investigate the effect and mechanism of gastrodin on head twitch behavior in rats with Tourette syndrome (TS). Methods 1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane (DOI) induction method was used to replicate TS rat model. Forty Wistar rats were randomly divided into four groups (n=10): the normal group (gastric administration of normal saline), the TS model group (from the first day of modeling, intragastric administration with normal saline at 2 h after DOI injection), the TS+ gastrodin group (from the first day of modeling, intragastric administration with 30 mg/kg gastrodin at 2 h after DOI injection), and the TS+gastrodin+CHIR-99021 group [glycogen synthase kinase-3β (GSK- 3β) inhibitor] (from the first day of modeling, 2 h after DOI injection, intraperitoneal injection with 2 mg/kg CHIR-99021 firstly, and then intragastric administration with 30 mg/kg gastrodin), continuous administration for 21 d. After the last administration, the rat’s head twitching behavior within 30 min was counted. The levels of striatal 5-hydroxyindole (5-HT), 5-hydroxyindole acetic acid (5-HIAA), and dopamine (DA) were detected by kits. The number of DA neurons in the substantia nigra was detected by immunohistochemical method. The expression of 5-HT transporter (SERT), 5- hydroxyindole 2A receptor (5-HT2AR), 5-hydroxyindole 2C receptor (5-HT2CR) and GSK-3β phosphorylation in the striatum were detected by Western blot assay. Results Compared with the normal group, the number of head twitches, DA level, number of DA neurons, SERT level and p-S9-GSK-3β, p-S9-GSK-3β/t-GSK-3β level increased in the TS model group, and the 5-HT level decreased (P＜0.05). Compared with the TS model group, the number of head twitches, DA level, number of DA neurons, SERT level and p-S9-GSK-3β, p-S9-GSK-3β/t-GSK-3β level decreased in the TS+gastrodin group, and the 5-HT level increased (P＜0.05). Compared with the TS+gastrodin group, the number of head twitches, DA level, number of DA neurons, SERT level and p-S9-GSK-3β, p-S9-GSK-3β/t-GSK-3β level increased , and the 5-HT level decreased in the TS+gastrodin+CHIR-99021 group (P＜0.05). Conclusion Gastrodin can improve the head twitch behavior of TS rats, and its mechanism may be by inhibiting the phosphorylation of GSK-3β, regulating the expression of SERT, affecting the level of 5-HT, and then inhibiting the release of DA in striatum. Related Articles | Metrics

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Effects of fasudil on oligodendrocyte maturation and myelination after white matter injury in preterm rats #br# TANG Yinglin, XU Qiongfang, XU Liting 2022, 50 (2):  160-166.  doi: 10.11958/20203641 Abstract ( 380 )   PDF (1399KB) ( 2537 )   Objective To investigate the effects of fasudil (FSD) on oligodendrocyte (OLs) maturation and myelination after white matter injury (WMI) in preterm rats. Methods Eighty male pups on postnatal day 2 (P2) were randomly divided into the sham group (n=30), the WMI group (n=25) and the FSD group (n=25). WMI model was established in the WMI group and the FSD group. After modeling, the FSD group received intraperitoneal injection of FSD［25 mg/(kg·d)］for 4 consecutive days. The WMI group and the sham group were given the same amount of saline. Neurofunctional severity score was used to evaluate the neurofunctional severity. TUNEL assay was used to determine the apoptosis of OLs, and immunofluorescence was performed to detect the expression levels of Olig2, Nkx2.2 and myelin basic protein (MBP). Western blot assay was used to detect the expression of MBP. The ultrastructure of corpus callosum and corona radiata was observed by electron micrograph, and the small excited postsynaptic current (mEPSC) was analyzed by electrophysiology in vitro. The balance test and the new object recognition test were used to evaluate the motor behavior of rats. Results The neurological severity score was significantly lower in the FSD group than that of the WMI group at 0.5, 1, 2 and 4 days postoperatively (P＜0.05). FSD treatment significantly decreased the density of total apoptotic cells and OLs apoptotic cells in corpus callosum and corona radiata (P＜0.05), and increased the density of Olig2+Nkx2.2+ cells and MBP volume fraction (P＜0.05). In addition, the number of myelinated axons in the corpus callosum and corona radiata of the WMI group was less than that of the sham group, while the FSD group had more myelinated axons than WMI group (P＜0.05). In vitro electrophysiological analysis showed that the frequency of mEPSC was significantly higher in the FSD group than that of the WMI group (P＜0.05). Behavioral test results showed that the frequency of foot slip was lower in the FSD group than that in the WMI group, and which showed better recognition ability. Conclusion FSD can induce OLs maturation, promote myelination and neural function recovery, and can be used as a feasible drug for the treatment of WMI in premature infants to promote clinical recovery. Related Articles | Metrics

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Correlation study between the SCF and SDF-1 in follicular fluid and embryonic development of POI #br# LIU Baoling, ZAHNG Jing, ZHONG Xuemei, JIN Xuan 2022, 50 (2):  167-171.  doi: 10.11958/20211583 Abstract ( 354 )   PDF (597KB) ( 2582 )   Objective To analyze the correlation between stem cell factor (SCF) and stromal cell-derived factor-1 (SDF-1) in follicular fluid of patients with primary ovarian insufficiency (POI), and the correlation between SCF and embryonic development. Methods A total of 144 infertile patients who underwent routine in vitro fertilization (IVF) treatment in our center were selected. According to the diagnosis, they were divided into the POI group (n=44), the elderly group (≥ 40 years old, n=40) and the normal group (< 40 years old, n=60). The levels of SCF, SDF-1 and E2 in follicular fluid were detected by ELISA. The correlation between SCF, SDF-1 and E2 was analyzed in different groups. The POI group was divided into three subgroups according to the SCF concentration. The differences in normal fertilization rate, cleavage rate and excellent embryo rate were compared between POI subgroups. Results The SCF level of follicular fluid was significantly higher in the POI group and the elderly group than that in the normal group (P＜0.05). SDF-1 and E2 were significantly lower in the POI group than those in the normal group (P＜0.05). SCF was negatively correlated with SDF-1 (rs= -0.400，P＜0.05）and positively correlated with E2 (rs=0.341，P＜0.05) in follicular fluid of the POI group. In the POI subgroups, the excellent embryo rates of the ＜200 ng/L group and the 200-300 ng/L group were significantly lower than those of the > 300 ng/L group (P＜0.05). Conclusion SCF and SDF-1 may interact with each other to participate in the development of POI, and SCF may be an valuable biochemical marker to predict embryonic development. Related Articles | Metrics

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Study on the correlation between serum neutrophil extracellular trap level and lupus nephritis #br# HAN Jing, GU Jinyi, LIU Yi, QIN Yuan, ZHAO Linghua 2022, 50 (2):  171-176.  doi: 10.11958/20211707 Abstract ( 399 )   PDF (449KB) ( 2673 )   Objective To investigate the relationship between serum neutrophil extracellular traps (NETs) and the prognosis of lupus nephritis (LN), and to analyze the prognostic value of NETs in LN. Methods There were 92 patients with LN (the LN group), 97 patients with systemic lupus erythematosus (SLE) and normal renal function (the SLE group), and 83 healthy volunteers (the control group) in this study. According to SLEDAI-2000 score, LN patients were divided into the mild activity group (score≤6, n=31), the moderate activity group (score7-12, n=33), the severe activity group (score＞12, n= 28). According to the occurrence of renal endpoint events during the follow-up period, LN patients were divided into the poor prognosis group (n=25) and the good prognosis group (n=67). Levels of serum NETs were detected to analyze the correlation between NETs and deoxyribonuclease 1 (DNASE1), renal function, SLEDAI-2000 score, renal tissue activity index (AI) and chronic index (CI) scores. Logistic regression was used to analyze the factors affecting the prognosis of LN, and receiver operating characteristic curve (ROC) was used to analyze the prognosis value of NETs in patients with LN. Results The serum level of NETs was higher in the LN group than that of the SLE group and the control group. The serum level of NETs was higher in the severe activity group than that of the moderate activity group and the mild activity group. The serum NETs level was higher in the moderate activity group than that of the mild activity group (P＜0.05). Levels of NETs were negatively correlated with glomerular filtration rate (eGFR) and DNASE1 (r=-0.769), and positively correlated with 24 h urinary protein quantification, SLEDAI-2000 score, renal tissue AI and CI (P＜0.05). High AI score and high NETs were risk factors for poor prognosis of LN (P＜0.05). The area under curve (AUC) of NETs for predicting prognosis in patients with LN was 0.871, with a sensitivity of 80.00% and a specificity of 86.57%. Conclusion Elevated levels of NETs are associated with degree of renal injury and renal end point events in LN, and can be used as a prognostic indicator for LN. Related Articles | Metrics

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Clinical study of cardiovascular protective effect of Qishen yiqi dropping pills on patients with hypertension complicated by coronary heart disease #br# ZHANG Ying, SHAO Dujing, LIU Xiaogang, ZHAO Yingqiang, LIU Yujie 2022, 50 (2):  177-181.  doi: 10.11958/20212082 Abstract ( 1239 )   PDF (422KB) ( 2739 )   Objective To observe the influences of Qishen yiqi dropping pills on blood pressure, blood lipid, inflammatory factors, anginal symptoms and major adverse cardiovascular events in patients with hypertension complicated by coronary heart disease. Methods Data of 192 patients with hypertension complicated by coronary heart disease were retrospectively analyzed. Patients were randomly divided into two groups, 96 for each group. The control group were given standard treatment of coronary artery disease and hypertension. The experimental group were given Qishen yiqi dropping pills (0.5 g once, three times/d) combined with standard treatment of coronary artery disease and hypertension. Changes of systolic blood pressure (SBP), diastolic blood pressure (DBP), total cholesterol (TC), low- density lipoprotein cholesterol (LDL-C), C-reactive protein (hs-CRP), interleukin 6 (IL-6), scores of symptom dimensions of Seattle angina pectoris scale (SAQ) including physical limitation, anginal stability, anginal frequency, treatment satisfaction and the incidence rate of major adverse cardiovascular events before and after treatment were compared between the two groups. Results Data of SBP, DBP, TC, LDL-C, hs-CRP and IL-6 were significantly decreased after treatment compared with that of before treatment (P＜0.01), and scores of physical limitation, anginal stability, anginal frequency and treatment satisfaction were significantly increased after treatment in the 2 groups (P＜0.05). After 1 year treatment, SBD, DBP, the levels of TC, LDL-C, hs-CRP and the incidence rate of major adverse cardiovascular events were significantly lower in the experimental group than those of the control group (P＜0.05). The scores of physical limitation, anginal stability, anginal frequency and treatment satisfaction were significantly higher in the experimental group than those of the control group (P＜0.01). Conclusion For patients with hypertension complicated by coronary heart disease, Qishen yiqi dropping pills can play a more effective role in lowering blood pressure, reducing levels of blood lipid and inflammation, and improving anginal symptoms and consequently reducing incidence rate of major adverse cardiovascular events, on the basis of standard Western medicine therapy. Related Articles | Metrics

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Effects of ketamine and lidocaine on free radical production after tournique-induced ischemiareperfusion injury in patients undergoing knee arthroplasty #br# FAN Fei, LEI Bibo 2022, 50 (2):  181-185.  doi: 10.11958/20211610 Abstract ( 400 )   PDF (404KB) ( 2565 )   ObjectiveTo evaluate the effects of different doses of ketamine and lidocaine on free radical production after tournique-induced ischemia-reperfusion injury in patients undergoing knee arthroplasty. Methods A total of 120 patients who planned to undergo knee arthroplasty were included in this study. Patients were randomly divided into the lidocaine group, the ketamine group and the saline group, with 40 patients in each group. The lidocaine group was given lidocaine intravenously with 1 mg/kg 10 min before anesthesia induction. In the ketamine group, 0.5 mg/kg ketamine was slowly injected intravenously 10 min before anesthesia induction. The saline group was intravenously injected with 0.9% sodium chloride 10 mL. Data of systolic blood pressure (SBP), diastolic blood pressure (DBP) and heart rate (HR) of three groups were recorded before anesthesia (T0), 5 min after infusion of intervention drugs (T1), 30 min after applying tourniquet (T2), and 15 min after removing tourniquet (T3). Peripheral venous blood samples were collected at T0, T2 and T3 to detect lactic acid level and ischemia modifier albumin (IMA) level in the three groups. The time of extubation, incidence of intraoperative hypotension, intraoperative hypertension, postoperative nausea and vomiting, agitation during recovery, delayed awakening and other adverse reactions were recorded. ResultsA total of 113 patients were enrolled, including 38 in the lidocaine group, 38 in the ketamine group, and 37 in the saline group, respectively. At T1, SBP, DBP and HR were higher in the ketamine group than those in the lidocaine group and saline group (P＜0.05). There were no significant differences in SBP, DBP and HR between the lidocaine group and the saline group (P＞0.05). At T2, SBP and DBP were significantly higher in the ketamine group than those in the lidocaine group and the saline group (P＜0.05), but there was no significant difference in HR between the two groups (P＞0.05). At T3, there were no significant differences in SBP, DBP and HR between the 3 groups. At T2 and T3, the serum levels of lactic acid and IMA were significantly lower in the lidocaine group than those of the ketamine group and the saline group, while the level was significantly lower in the ketamine group than that of the saline group (P＜0.05). There were no significant differences in the time of extubation, intraoperative hypertension, incidence of intraoperative hypotension, postoperative nausea and vomiting, agitation in recovery period and delay in recovery between the three groups (P＞0.05). Conclusion Both lidocaine and ketamine can protect against tourniquet-induced ischemia-reperfusion injury in patients undergoing knee arthroplasty, but lidocaine is more effective. Related Articles | Metrics

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The expression of serum TXNIP in patients with type 2 diabetic retinopathy and its relationship with oxidative stress TAO Lin, ZHAO Xiaolian, QI Shufang 2022, 50 (2):  186-190.  doi: 10.11958/20211480 Abstract ( 529 )   PDF (438KB) ( 2578 )   Objective To investigate the expression of serum thioredoxin-interacting protein (TXNIP) in patients with type 2 diabetic retinopathy (DR) and its relationship with oxidative stress. Methods According to the severity of the disease, 82 DR patients (the DR group) were divided into the PDR group (42 cases) and the NPDR group (40 cases). In addition, 50 healthy volunteers (the control group) and 50 patients with simple type 2 diabetes (the diabetes group) were selected. General data of all subjects were collected. The levels of superoxide dismutase (SOD), reduced glutathione (GSH), malondialdehyde (MDA) and TXNIP were detected by xanthine oxidase method, dithio Dinitrobenzoic acid colorimetry, thiobarbital method and enzyme-linked immunosorbent assay. Glycated hemoglobin (HbA1c), fasting blood glucose (FPG), triglyceride (TG) and total cholesterol (TC) were determined by automatic biochemical analyzer. Results Compared with the control group, TG and TC increased significantly in the diabetes group and the DR group (P＜0.05), but there was no significant difference between the diabetes group and the DR group. HbA1c, FPG, MDA and TXNIP increased sequentially in the control group, the diabetes group and the DR group, while SOD and GSH decreased sequentially (all P＜0.05). Compared with the PDR group, MDA and TXNIP in the NPDR group decreased, while SOD and GSH increased (P＜0.05). There were no significant differences in HbA1c, FPG, TG and TC between the two groups. Higher levels of MDA and TXNIP were all risk factors for PDR in DR patients, higher levels of SOD and GSH were protective factors (P＜0.05). The area under ROC curve of serum TXNIP in the diagnosis of DR was 0.803 (95%CI: 0.730-0.875). When the cutoff value was 3.01 μg/L, the sensitivity, specificity and Youden index were 53.66%, 94.00% and 0.477, respectively. Conclusion TXNIP is abnormally high expressed in the serum of DR patients, and it may promote the occurrence and development of the disease by regulating oxidative stress response. High serum TXNIP level is a risk factor for PDR in DR patients, and serum TXNIP has certain auxiliary value in clinical diagnosis of DR. Related Articles | Metrics

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Effects of plasma vWF and vWF lyase levels on diabetic nephropathy CAO Lei, HU Enbi, LI Xiaonan 2022, 50 (2):  190-194.  doi: 10.11958/20211643 Abstract ( 535 )   PDF (435KB) ( 2606 )   Objective To explore the levels and their possible effects of serum von Willebrand factor (vWF) and vWF lyase (ADAMTS13) in patients with diabetic nephropathy (DN). Methods Fifty cases of healthy physical examination (the normal control group) and 144 patients with type 2 diabetes (the T2DM group) were selected in this study. According to the ratio of urine microalbumin to urine creatinine (ACR), the diabetic T2DM patients were divided into the simple diabetes group (n=42, ACR＜30 mg/g), the microalbuminuria group (n=56, 30 mg/g≤ACR˂300 mg/g), and the macroalbuminuria group (n=46, ACR≥300 mg/g). The immunoturbidimetric method was used to detect vWF levels. The double antibody sandwich method was used to detect ADAMTS13 level, creatinine (sCr), urea nitrogen (BUN) and glycosylated hemoglobin (HbA1c). The glomerular filtration rate (eGFR), D-Dimer and fibrinogen (Fib) were estimated. Results HbA1c, Fib, D-dimer and vWF increased successively in the normal control group, the T2DM group, the microalbuminuria group and the macroalbuminuria group, while ADAMTS13 decreased in turn (P＜0.05). Multiple linear regression analysis showed that albumin and D-dimer were the main factors affecting serum vWF, and D-dimer was the main factor affecting ADAMTS13 levels. ROC curve analysis showed that vWF was more sensitive in diagnosing DN renal damage, while D-dimer had more balanced sensitivity and specificity in the diagnosis of DN renal damage, and specificity was more advantage. Conclusion Serum vWF and ADAMTS13 may play an important role in the occurrence and development of DN, and their levels can be used as clinical indicators for the early diagnosis and prediction of DN. Related Articles | Metrics

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Effects of leg elevation and using exsanguination band blood-expelling methods on the short-term effectiveness after total knee arthroplasty ZHANG Yuying, JIANG Jun, MENG Xiaoguang, MA Peng, TANG Shusen, REN Qiuxiao 2022, 50 (2):  195-199.  doi: 10.11958/20212738 Abstract ( 460 )   PDF (428KB) ( 2622 )   Objective To compare and explore the effects of leg elevation of blood-expelling and blood-expelling by the exsanguination band on the early clinical outcomes during total knee arthroplasty (TKA). Methods A total of 80 patients with initial TKA were selected. According to different blood-expelling methods, patients were divided into the leg elevation of blood-expelling group (n=40) and the blood-expelling by the exsanguination band group (n=40). The clinical data were recorded. The levels of hemoglobin (HGB), creatine kinase (CK), C-reactive protein (CRP), interleukin (IL) -6, tumor necrosis factor (TNF)-α and D-Dimer 1 d before operation and 1 d after operation were compared between the two groups. The visual analogue scale (VAS) and knee-joint range of motion (ROM) 1 d before operation and 1 d, 3 d, 7 d and 1 month after operation, and Hospital for Special Surgery Score (HSS) 1 d before operation and 1 d, 7 d, 1 month and 3 months after operation were compared and analyzed between the two groups. The incidences of postoperative complications were recorded in the two groups. Results There were no significant differences in general data and clinical indexes before operation between the two groups. One day after operation, the level of HGB was higher in the leg elevation of bloodexpelling group than that in the other group (P＜0.05). The levels of CRP, CK, IL-6, TNF-α and D-Dimer were lower in the leg elevation of blood-expelling group than those in the blood-expelling by the exsanguination band group (P＜0.05). The knee-joint ROM and HSS of the two groups increased with time after operation, while the VAS decreased. The knee-joint ROM was higher in the leg elevation of blood-expelling group than that in the blood-expelling by the exsanguination group on 1 d and 3 d after operation, while the VAS were lower on 1 d, 3 d, and 7 d after operation in the leg elevation of bloodexpelling group than the another group (P＜0.05). There were no significant differences for HSS between different blood-expelling methods. The incidence of lower limb swelling after operation was higher in the blood-expelling by the exsanguination band group than that in the leg elevation of blood-expelling group (P＜0.05). The occurrence of other complications between the two groups were no significant differences (P＜0.05). Conclusion During TKA, leg elevation of blood-expelling method is not only simple to operate, but also has more advantages in reducing early pain, swelling, inflammatory response, vein thrombosis and other complications, which helps patients recover quickly has higher clinical practical value. Related Articles | Metrics

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The application value of region of interest selection for intravoxel incoherent motion diffusion-weighted imaging parameters in the evaluation of tumor angiogenesis and lymphovascular invasion in rectal cancer LIAO Zonghui , YUAN Yi, PU Hong, LI Hang 2022, 50 (2):  200-205.  doi: 10.11958/20211221 Abstract ( 690 )   PDF (616KB) ( 2521 )   Objective To evaluate the application value of region of interest (ROI) selection for intravoxel incoherent motion (IVIM) diffusion-weighted imaging parameters with MVD, VEGF and LVI in rectal cancer. Methods Fifty-four patients with rectal cancer underwent preoperative IVIM and conventional MRI were included in this study. Three methods of ROI measurements (volume ROI, maximum area ROI and small ROI) were used to independently measure the IVIM parameters including pure diffusion coefficient (D), perfusion-related diffusion coefficient (D*), and perfusion fraction (f) values of the rectal cancer. The expressions of VEGF, MVD and LVI were detected by immunohistochemical staining. IVIM parameters were analyzed in the different expressions of VEGF, MVD and LVI in rectal cancers. The receiver operating characteristic (ROC) curve of each parameter of IVIM was analyzed, and the area under the ROC curve (AUC) was calculated to evaluate the diagnostic performance of each parameter for VEGF, MVD and LVI. Results Among the three ROI methods used by 2 doctors to detect each parameter value, the consistency of D value, D* value and f value measured by volume ROI was the best, and the consistency of D value diagnosis among the three parameters of IVIM was generally more advantageous. Among the low and high MVD expressions, only the D value measured by the volume ROI showed a significant difference (P＜0.05). In VEGF negative and positive patients, only the volume ROI and small ROI measured D values were significant (P＜0.05). In LVI-negative and positive patients, only the D value measured by volume ROI had a statistically significant difference (P＜0.05). ROC curve analysis results showed that the D value of volume ROI was better for judging MVD, VEGF and LVI, especially for judging the sensitivity of high MVD expression. The D value of small ROI showed better specificity for diagnosing negative expression of VEGF. Conclusion D value can reflect the angiogenesis of rectal cancer. In particular, D value measured by volume ROI can more objectively and accurately evaluate the MVD, VEGF and LVI before surgery. Related Articles | Metrics

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A pilot study on the value of triggered non-contrast enhanced MRI angiography in the diagnosis of varicose veins of lower extremities XU Tuanxin, LIU Xiao, ZHU Gangming, WU Shaoteng, LIU Chengkang, YANG Gai, YE Peihao 2022, 50 (2):  205-209.  doi: 10.11958/20211670 Abstract ( 617 )   PDF (622KB) ( 2494 )   Objective To investigate the clinical application of triggered non-contrast enhanced (TRANCE) magnetic resonance imaging in varicose veins (VV) of lower extremities. Methods Thirty patients with 41 segments of veins were examined by TRANCE and Doppler ultrasonography (DUS). Pearson correlation analysis was used to analyze the signal ratio (SR) and vessel diameter ratio (VDR) of VV. The Likert score consistency of TRANCE images was evaluated between the two doctors and between before and after 2 weeks. The digital subtraction angiography (DSA) was used as the gold standard, the diagnostic efficiency and consistency of the two inspection methods were evaluated. Results SR was negatively correlated with VDR (r=-0.440，P＜0.05). The overall Likert score was lower in the pelvic segment than that of thigh segment (P＜ 0.05), and there was no significant difference in Likert scores between the thigh and calf segments. The two doctors had good consistency in Likert scores of the pelvic, thigh, and calf segments before and after 2 weeks, and the Likert scores elevated by the same doctor before and after 2 weeks. With surgical results as the gold standard, the sensitivity of TRANCE and DUS of 41 segments of VV were 94.59% and 86.49%, the specificity were 50.00% and 75.00%, and the accuracy were 90.24% and 85.37%, respectively. The diagnostic efficiency kappa value of the two methods was 0.617, indicating good consistency. Conclusion TRANCE can quantitatively analyze the degree of VV and has good image quality in the diagnosis of lower extremity varicose veins, and the diagnostic efficiency is consistent with DUS. It can be used as an important preoperative auxiliary examination method. Related Articles | Metrics

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The diagnostic value of abdominal ultrasound in the early differentiation of neonatal food protein-induced enterocolitis syndrome and neonatal necrotic enterocolitis YANG Tianjiao, KE Chuanghong, ZENG Cizheng, LIN Liman, HU Hailing, MO Xiaohuan 2022, 50 (2):  210-213.  doi: 10.11958/20211732 Abstract ( 429 )   PDF (572KB) ( 2531 )   Objective To explore the value of abdominal ultrasound in the differential diagnosis of neonatal food protein-induced enterocolitis syndrome (NFPIES) and early neonatal necrotic enterocolitis (NEC). Methods Twenty-one children diagnosed as NFPIES and early NEC (n=48) were selected in this study. Data of sex, gestational age, clinical symptoms (vomiting, abdominal distension and bloody stool) and the percentage of blood eosinophils (EO) within 8 hours of clinical symptoms were collected. The time of the first abdominal ultrasound examination of all children was within 8 hours of the occurrence of clinical symptoms, and the interval between the abdominal ultrasound examination and the first examination was 8-24 hours. The changes of abdominal ultrasound signs (intestinal wall pneumatosis, intestinal peristalsis, intestinal wall thickening and peritoneal effusion) were dynamically compared. Results There were no significant differences in general baseline data between the NFPIES group and the early NEC group. The proportion of peripheral blood EO and the proportion of children with EO ratio≥0.05 were significantly higher in the NFPIES group than those in the early NEC group (P＜0.01). The proportion of children with positive abdominal ultrasound signs was lower in the NFPIES group than that in the early NEC group (P＜0.05). The ultrasonographic signs of peristalsis showed slowing at the diseased site, and the remaining peristalsis was normal without peristalsis disappearance or peritoneal effusion. In the re-examination of abdominal ultrasound, the ultrasound signs of gas in the intestinal wall and slowing of bowel movement were significantly lower in the NFPIES group than those with positive abdominal ultrasound examination for the first time (P＜0.05), while there was no significant difference in the proportion of four children with positive abdominal ultrasound signs in the early NEC group compared with the first examination. In the re-examination of abdominal ultrasound, the proportion of four children with positive ultrasound signs was lower in the NFPIES group than that in the early NEC group (P＜0.01). Conclusion Abdominal ultrasound signs help to distinguish NFPIES from early NEC, and dynamic observation of its changes is of greater significance. Related Articles | Metrics

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Recent advances in the mechanisms of drug resistance and treatment of BRAF-mutant melanoma LI Xiyuan, ZHU Zhixin, ZHAO Hailong 2022, 50 (2):  214-219.  doi: 10.11958/20212135 Abstract ( 463 )   PDF (427KB) ( 2500 )   Melanoma is the primary malignant tumor of the skin with a very high morbidity and fatality, of which 50% are caused by viral infection of proto-oncogene BRAF mutation. The combination of BRAF and MEK inhibitors is the main treatment of BRAF mutant melanoma, but there have been some drug resistance and drug side effects. Currently, resistance of BRAF mutant melanoma is considered to include primary resistance, adaptive resistance and secondary resistance, and corresponding molecular mechanisms are related and different. At the same time, in order to reduce the incidence of drug resistance in BRAF mutant melanoma, some progress has been made in basic research and clinical application of new target inhibitors, immunotherapy and epigenetics. In this paper, we will summarize the possible mechanism of drug resistance in BRAF mutant melanoma and the current situation and prospects of optimized treatments. Related Articles | Metrics

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Progress in clinical trials of PD-1 inhibitor monoclonal antibody for nasopharyngeal carcinoma LEI Xiaomei, QU Jiaquan, TAN Tan 2022, 50 (2):  220-224.  doi: 10.11958/20211672 Abstract ( 397 )   PDF (389KB) ( 2457 )   Nasopharyngeal carcinoma (NPC) is a head and neck malignant tumor with high infection rate of EBV. The activation of programmed death protein-1 and its ligand (PD-1/PD-L1) pathway may be one of the mechanisms of immune escape from EBV-associated NPC. Compared with chemotherapy, immune checkpoint inhibitors have less adverse reactions. Survival benefit is longer and the advantages of the tolerance is better, but there is only a small number of patients will benefit from faults. At present, at least four kinds of inhibitors targeting the pathway of PD-1 single resistance in clinical trials have achieved good results, and it has been approved for the treatment of recurrent or metastatic nasopharyngeal carcinoma. Recent completed and ongoing studies on the PD-1 inhibitor treatment strategy for NPC, the efficacy evaluation and safety studies were reviewed. Related Articles | Metrics