Table of Content

05 February 2024, Volume 44 Issue 2

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Original Articles

Sestrin1 is involved in the regulation of gluconeogenesis in mouse liver cells

GUO Yanfang, GENG Chao, XIE Xianghong, CHEN Enhui, GUO Zeyu, ZHANG Minglong, LIU Xiaojun

2024, 44(2):  141-146.  doi:10.16352/j.issn.1001-6325.2024.02.0141

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Objective To investigate the role and regulatory mechanism of stress-inducing protein 1(SESN1) in liver gluconeogenesis of fasting mice. Methods RT-qPCR was used to detect mRNA expression of SESN1 in liver tissues of C57BL/6J mice and primary mouse hepatocytes treated with forskolin (Fsk) and dexamethasone(Dex). HepG2 cells were transfected with plasmids and the effects of SESN1 overexpression on mRNA expression of gluconeogenesis related genes PGC-1α, PEPCK and G6Pase was detected by RT-qPCR. The effect of SESN1 on the promoter activity of PGC-1α in HepG2 cells was studied using a dual luciferase reporter system. The effect of SESN1 on PGC-1α deacetylation was detected by overexpression of SESN1 and inhibition of SIRT1 expression.By knocking down SIRT1 expression, we detected whether it mediated the changes in mRNA levels of SESN1 induced gluconeogenesis related genes. Results The mRNA expression of SESN1 was significantly increased in liver tissues of starved C57BL/6J mice and in primary hepatocytes treated with Fsk and Dex(P＜0.001). Over-expression of SESN1 in HepG2 cells promoted mRNA expression of PGC-1α, PEPCK and G6Pase(P＜0.001) and promoter activity of PGC-1α(P＜0.001). Over-expression of SESN1 decreased the acetylation level of PGC-1α in primary hepatocytes.Sirt family inhibitors NAM and shRNA adenovirus interfered with SIRT1 expression respectively, and antagonized the deacetylation effect of SESN1 on PGC-1α. The expression of PGC-1α, PEPCK and G6Pase induced by SIRT1 was also significantly impaired(P＜0.000 1). Conclusions SESN1 regulates liver gluconeogenesis in mice with a SIRT1-dependent mechanism.

Dexmedetomidine alleviates TNF-α-induced injury of human neuroblastoma cell line SH-SY5Y through activating AMPK

MU Yang, YANG Zhiwen

2024, 44(2):  147-153.  doi:10.16352/j.issn.1001-6325.2024.02.0147

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Objective To study the effect of dexmedetomidine (DEX) on TNF-α-induced injury of human neuroblastoma cell line SH-SY5Y and its mechanism. Methods CCK8 assay was used to detect cell activity and determine the optimal dose of DEX and TNF-α. The cells were divided into control group, model group, DEX intervention model group, and the intervention group of DEX plus compound C. Western blot was used to detect the expression of p-AMPK, SNHP, KIF5B, Drp1 and OPA1, and ELISA was used to detect the level of IL-1β and IL-6. Mitochondrial membrane potential (Δψm), respiratory chain complex enzyme activity (complex Ⅰ-Ⅳ), ATP, MDA, SOD, GSH, ROS were determined with commercially available kits. Results Compared with control group, level of p-AMPK,OPA1 and SNPH l in model group significantly decreased, while the level of Drp1 and KIF5B significantly increased (P＜0.01). The level of complex Ⅰ~Ⅳ, Δψm, ATP, GSH and SOD in DEX group significantly increased as compared with model group. The level of MDA, IL-1β and IL-6 significantly decreased(P＜0.01). Compared with DEX group, the level of Complex Ⅰ-Ⅳ, Δψm, ATP, GSH and SOD in DEX+CC group significantly decreased, while the level of MDA, IL-1β and IL-6 significantly increased(P＜0.01). Conclusions DEX improves mitochondrial function, alleviates oxidative stress and inflammatory response as well as TNF-α-induced cell damage with an AMPK dependent mechanism.

Nutlin-3a regulates mouse adipose function by inhibition of CIDEC expression

CHEN Enhui, YANG Jiahui, ZHAO Wei, XIE Xianghong, GUO Yanfang, LIU Xiaojun, YAN Li

2024, 44(2):  154-158.  doi:10.16352/j.issn.1001-6325.2024.02.0154

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Objective To investigate the effect of Nutlin-3a, a mouse double minute 2 homolog (MDM2) inhibitor, on lipid metabolism of mouse adipose. Methods High-fat diet-induced obesity (DIO) C57BL/6J mice were randomly divided into a control group injected with DMSO and an experimental group injected with Nutlin-3a. Then we conducted glucose tolerance (GTT) and insulin tolerance (ITT) tests. The epididymal white adipose tissue (eWAT), inguinal white adipose tissue (iWAT) and brown adipose tissue (BAT) of animals were isolated and microscopy of WATs with hematoxylin-eosin (HE) staining was performed to observe the morphological changes of adipocytes. The expression of lipid metabolism related gene cell death-inducing DFF45-like effector C (CIDEC) in eWAT were detected by qPCR and Western blot. Results Compared with the control group, Nutlin-3a was found to promote the body weight (P＜0.001),but no effect on glucose tolerance and insulin sensitivity in DIO mice. Nutlin-3a treatment decreased the size of adipocytes and fat deposition in adipose tissue and downregulated the mRNA and protein levels of CIDEC in eWAT. Conclusions Nutlin-3a inhibits the formation of lipid droplets by downregulating expression of CIDEC in white adipose tissue.

Optimisation of primary osteoblast cell culture from suckling mouse

WANG Zuoyu, ZHOU Yang, YANG Junwei

2024, 44(2):  159-166.  doi:10.16352/j.issn.1001-6325.2024.02.0159

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Objective To develop a suitable medium and optimize culture time for the primary osteoblast culture from suckling mouse, so to provide an improved experimental protocol for primary osteoblast culture in vitro. Methods Primary osteoblasts were collected from skull of CD1 suckling mouse by interrupted enzyme digestion. The purified osteoblasts were harvested by differential centrifugation. The incubation time, concentration of fetal bovine serum(FBS),β-glycerophosphate sodium and dexamethasone were tested and optimized. The change of osteoblast maturation marker was examined by Western blot(WB) and immunofluorescence staining (IF). The osteogenic activity was determined by alkaline phosphatase staining, alizarin red staining and ultrastructure. Results Primary osteoblast were obtained from sucleling mouse skull bone by interrupted enzyme digestion for proliferation and transgenerational expansion.The expression of osteoblast maturation markers was parallel to the time of induction culture and the concentration of FBS. Mature osteoblasts were obtained by culturing the cells with 10% FBS for 14 days. The differentiation of primary osteoblasts was induced by different concentrations of β-glycerophosphate and dexamethasone. The results showed that the expression of osteoblast maturation markers was higher under the culture conditions of 10 mmol/L β-glycerophosphate and 5 nmol/L dexamethasone(P＜0.01), and the staining of alkaline phosphatase and alizarin red was obvious,and the osteogenic activity was better too. Conclusions Primary osteoblasts isolated from the skull of suckling CD1 mice cultured in induction medium containing 10% fetal bovine serum, 10 mmol/L β-glycerophosphate sodium and 5 nmol/L dexamethasone for 14 days show good osteogenic activity and are suitable for in vitro experimental studies.

Expression and role of triggering receptor expressed on myeloid cells 2 in high glucose-treated microglia

WANG Zhaohui, LIU Xiao, ZHOU Yue, WEI Xinyi, WANG Yue, LI Junfa, ZHAO Li

2024, 44(2):  167-173.  doi:10.16352/j.issn.1001-6325.2024.02.0167

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Objective To explore the expression of triggering receptor expressed on myeloid cells 2 (TREM2) in high-glucose microglia and to investigate the role of TREM2 in the proliferation, migration and phagocytosis of high-glucose microglia. Methods Microglia cells were divided into control group and high-glucose treatment group(67.5 mmol/L glucose, 24 h). The microglia cells were counted and the expression of Iba1 and TREM2 was detected. TREM2 siRNA was transfected to detect the proliferation and migration of microglia. The amyloid β-peptide (Aβ) with a fluorescent tag was added to observe the phagocytosis of Aβ by microglia. Results Compared to normal microglia, the number of microglia significantly decreased after high-glucose treatment (P＜0.001), while the expression of TREM2 and Iba1 markedly increased (P＜0.001). High glucose and TREM2 did not affect the proliferation of microglia. Compared to the normal group, the migration of microglia significantly decreased after high-glucose treatment (P＜0.05) and TREM2 did not affect the migration ability of high-glucose microglia. Compared to the normal group, the phagocytosis of Aβ by microglia significantly decreased in the high-glucose treated group(P＜0.001). Furthermore, TREM2 knockdown further decreased the phagocytosis of Aβ by high-glucose microglia (P＜0.001). Conclusions The expression of TREM2 in microglia significantly increases after high-glucose treatment, which significantly affects phagocytosis of Aβ by microglia.

Tetrandrine attenuates IL-1β-induced injury of human articular chondrocytes in vitro

OUYANG Yunlang, ZHANG Ning, CHEN Liping, JIA Bingshen, JIAO Tuo

2024, 44(2):  174-179.  doi:10.16352/j.issn.1001-6325.2024.02.0174

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Objective To investigate the effect of tetrandrine (Tet) on injury of primary human articular chondrocytes induced by interleukin-1β (IL-1β). Methods Human articular chondrocytes were divided into control group, IL-1β group, hypoxia inducible factor(HIF-1α) inhibitor group [2-methoxyestradiol(2-ME2) group], Tet groups containing low, medium and high concentrations. Each group has six replicated samples. MTT assay was used to detect the viability of cells; cell apoptosis was detected by flow cytometry; the level of inflammatory related factors like tumor necrosis factor-α (TNF-α), matrix metalloproteinase 3(MMP-3), inducible nitric oxide synthase (iNOS), cyclooxygenase-2(COX-2) and the activity of antioxidant factors like super oxide dismutase (SOD) and glutathione peroxides (GPx) in cells were detected by ELISA; Western blot was used to detect the expression of HIF-1α and VEGF in cells. Results Compared with the control group, the apoptosis rate, level of TNF-α,MMP-3, iNOS,COX-2 and the protein expression of HIF-1α and VEGF in IL-1β group all increased, the cell survival rate and the activity of SOD and GPx significantly decreased (P＜0.05); compared with IL-1β group, the apoptosis rate, the level of TNF-α, MMP-3, iNOS, COX-2 and the protein expression of HIF-1α and VEGF in 2-ME2 group and Tet groups with low, medium, and high concentration significanthy decreased(P＜0.05). The cell survival rate and the activity of SOD and GPx significantly increased(P＜0.05). Conclusions Tetrandrine attenuates IL-1β-induced injury of human articular chondrocytes.

Serum CA211 level and NK cell number are associated with prognosis of patients with non-small cell lung cancer

DING Fei, WANG Sa, HUANG Changxin

2024, 44(2):  180-184.  doi:10.16352/j.issn.1001-6325.2024.02.0180

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Objective To exploring the correlation between the expression level of serum carbohydrate antigen 211 (CA211) and the number of natural killer (NK) cells and prognosis in non-small cell lung cancer (NSCLC) patients. Methods 132 NSCLC patients admitted to the Affiliated Hospital of Hangzhou Normal University from June 2019 to July 2022 were selected as the test group, and 132 patients with benign lung lesions during the same period were selected as the control group. Data were collected from the laboratory to analyze the relationship between serum CA211 expression and NK cell count in NSCLC with clinical prognosis, as well as the related factors affecting the prognosis of NSCLC patients. Results The neutrophil to lymphocyte ratio(NLR) and serum CA211 expression in the test group were higher than those in the control group(P＜0.05), while the count of NK cells was less than that in the control group(P＜0.05); The expression level of serum CA211 in the test group was negatively correlated with the count of NK cells(r=-0.405, P＜0.001); There were significant differences in lymph node metastasis and TNM staging among patients with different levels of serum CA211 expression and NK cell count(P＜0.05); The one-year survival rate of patients with low expression of CA211 was significantly higher than that of patients with high expression of CA211(P＜0.05), and the one-year survival rate of patients with high count of NK cells was higher than that of patients with low count of NK cells(P＜0.05); Lymph node metastasis, TNM staging and CA211 level were all risk factors affecting the prognosis of NSCLC patients(P＜0.05), while counting of NK cells was protective factor for the prognosis of NSCLC patients(P＜0.05). Conclusions The level of serum CA211 and the number of NK cells in NSCLC patients are closely related to pathological characteristics and clinical prognosis.

Downregulation of demethylase FTO inhibits proliferation of human liver cancer cell line HepG2

LU Huiying, WANG Jianguo

2024, 44(2):  185-191.  doi:10.16352/j.issn.1001-6325.2024.02.0185

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Objective To explore the mechanism of the demethylase fat mass and obesity associatal(FTO) gene on the proliferation of human liver cancer cell line HepG2. Methods HepG2 cells were divided into the control group, FTO group (transfected with FTO over-expression plasmid), si-FTO group (transfected with si-FTO)and si-FTO+si-FoxO1 group (simultaneously transfected with si-FTO and si-FoxO1). The expression of FTO in cells was detected by RT-qPCR. Cell proliferation, invasion and apoptosis were examined using CCK-8 assay, Transwell chamber assay and flow cytometry, respectively. Dot blot assay was performed to measure m⁶A methylation , and protein expression of FoxO1 in cells was detected by Western blot. Results Analysis of overall survival in liver cancer patients using The Cancer Genome Atlas (TCGA) showed higher expression of FTO associated with shorter survival (P＜0.05). Compared with normal liver cells HL7702, FTO relative expression was significantly increased in human liver cancer cells HepG2 (P＜0.05).The relative expression of FTO in si-FTO group cells was lower than that in the control group, while the relative expression of FTO in FTO group was higher than that in the control group (P＜0.05). The relative expression of FTO in the si-FTO+si-FoxO1 group was higher than that in the si-FTO group(P＜0.05). Compared with the Control group, cell viability, count of invasive cells, relative level of m⁶A were significantly decreased, while apoptosis rate and protein expression of FoxO1 were significantly increased in the si-FTO group(P＜0.05). Cell viability, count of invasive cells, and relative expression level of m⁶A were significantly higher, while apoptosis rate and protein expression of FoxO1 were significantly lower in the FTO group compared to the Control group (P＜0.05). Compared with the si-FTO group, cell viability, invasive cells and relative level of m⁶A were significantly increased, while apoptosis rate and protein expression of FoxO1 were significantly decreased in the si-FTO+si-FoxO1 group(P＜0.05). Conclusion High expression of FTO is associated with poor clinical prognosis. Knockdown of the demethylase FTO gene inhibits proliferation and invasion of liver cancer cells and induces their apoptosis. The mechanism is potentially related to the regulation of FoxO1.

Guanxinning tablet alleviates carotid atherosclerotic plaque injury in rats

ZHANG Xiuli, HU Xiaogang, MO Jun, CHEN Ling

2024, 44(2):  192-198.  doi:10.16352/j.issn.1001-6325.2024.02.0192

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Objective To investigate the effect and its mechanism of Guanxinning tablet on carotid atherosclerotic plaque in a rat model. Methods The rats were randomly divided into control group, model group (to construct carotid atherosclerosis plaque model), Guanxinning groups with low, medium and high dose of Guanxinning tablet (150,300 and 600 mg/kg), atorvastatin group (2 mg/kg), lithiumchloridemonohydrate(LiCl) (Wnt/β-catenin pathway activator) group (15 mg/kg), and Guanxinning plus LiCl group (600 mg/kg Guanxinning tablets+15 mg/kg LiCl), with 12 rats in each group. The intervention began at the third week and was administered once a day for 8 weeks. Olympus Au2700 automatic biochemical analyzer was used to detect the level of total cholesterol (TC), triglyceride (TG), low density lipoprotein (LDL) and high-density lipoprotein (HDL) in rat serum; ELISA was applied to detect the level of monocyte chemotactic protein (MCP-1) and hyper-sensitive C-reactive protein (hs-CRP) in rat serum; the level of oxidized low density lipoprotein (ox-LDL) and malondialdehyde (MDA) in rat serum were detected by spectrophotometry; HE staining was applied to find pathological changes in the common carotid artery of rats; Western blot was applied to detect the expression of Wnt3a, matrix metalloproteinase-9 (MMP-9) and β-catenin in rat common carotid artery. Results Compared with the control group, the level of TG, TC, LDL, MCP-1, hs-CRP, ox-LDL, protein expression of MDA, MMP-9, Wnt3a, β-catenin and total plaque area/total arterial area ratio increased, the HDL level decreased in model group (P＜0.05); compared with the model group, the level of TG, TC, LDL, MCP-1, hs-CRP, ox-LDL, MDA, expression of MMP-9, Wnt3a, β-catenin protein and total plaque area/total arterial area ratio in the low, medium, and high dose groups of Guanxinning tablets decreased, the HDL level increased. The effect of Guanxinning tablets was dose-dependent, and the change trend of corresponding indicators in the LiCl group was opposite to the above (P＜0.05); compared with the high dose group of Guanxinning tablets, the TG, TC, LDL, MCP-1, hs-CRP, ox-LDL, MDA levels, MMP-9, Wnt3a, β-catenin protein expression, and total plaque area/total arterial area ratio in the high dose+LiCl group of Guanxinning tablets increased, the HDL level decreased(P＜0.05). Conclusions Guanxinning tablet can inhibit the formation of atherosclerotic plaque in rats and the mechanismis potentially related to the regulation of Wnt/β-catenin pathway.

Expression of junctophilin 2 and FGF23 in atrial tissue of rabbits with atrial fibrillation cardiomyopathy

GUO Shuang, LI Shuren, ZHAO Mei, HAO Xiao

2024, 44(2):  199-203.  doi:10.16352/j.issn.1001-6325.2024.02.0199

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Objective To explore the expression of junctophilin 2(JP2)and fibroblast growth factor 23(FGF23)in a rabbit model of atrial fibrillation mediated-cardiomyopathy(AMC). Methods Rabbit models of atrial fibrillation (AF) were developed through rapid atrial stimulation and then divided into three groups: control group (pacemakers implanted without pacing, n=6), AF group (pacing with ejection fraction decrease ＜10%,n=5), and AMC group (pacing with ejection fraction decrease ≥10%, n=6).Echocardiography was performed to detect left ventricular end-diastolic diameter(LVEDD),left ventricular end-systolic diameter(LVESD) and left ventricular ejection fraction(LVEF). JP2 and FGF23 were detected by ELISA method.Western blot and RT-qPCR were conducted to detect protein and mRNA expression of JP2 and FGF23. Results Left atrial diameter, right atrial diameter and right ventricular diameter increased and LVEF decreased in the AMC group as compared with the control group. AMC group had lower LVEF and larger aorta and right ventricle diameter. Compared with the control group, the expression of FGF23(P＜0.001)and JP2(P＜0.01)in left atrial cardiomyocytes was significantly increased in the AF group, while the expression of JP2 was decreased in the AMC group(P＜0.001).AMC group had lower expression of JP2 and FGF23 compared with AF group. Compared to the control group,plasma concentration of JP2 and FGF23 increased in the AF group and FGF23 plasma concentration increased in the AMC group. Plasma concentration of FGF23 and JP2 was lower in AMC group than that in AF group . Conclusions FGF23 expression increased and JP2 expression decreased as found in the rabbit AMC model.

FHL2 regulates THP-1 macrophage foaming through NF-κB signaling pathway

CHEN Weiwei, LIAO Huang, SHI Zhenhong, LUO Ying

2024, 44(2):  204-209.  doi:10.16352/j.issn.1001-6325.2024.02.0204

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Objective To determine whether four-and-a-half LIM-only protein 2(FHL2) can affect macrophage foaming by regulating nuclear factor kappa-B(NF-κB) signaling pathway. Methods FHL2 over-expression plasmids and siRNA of FHL2 were constructed and transfected into human monocyte/macrophages cell line THP-1. Western blot was used to detect the expression of FHL2. The cells were stimulated with oxidized low density lipoprotein(ox-LDL) and the expression of IL-6, IL-1β,TNF-α and other cytokines were detected by ELISA. Oil red O staining was used to detect the degree of cell foaming. The protein expression of NF-κB signaling pathway was detected by Western blot. Results The expression of FHL2 increased after transfected with FHL2 over-expression plasmids while decreased in si-FLH2 transfected cells. FHL2 down-regulated secretion of inflammatory cytokines. Down-regulation of FHL2 alleviated THP-1 macrophage foaming. The down-regulation of FHL2 inhibited activation of NF-κB signaling pathway, while the over-expression FHL2 showed an opposite trend. Conclusions FHL2 down-regulation inhibits the activation of NF-κB signaling pathway, reduces the secretion of inflammatory cytokines and alleviates foaming of macrophages.

CircRERE promotes proliferation, migration and invasion of human multiple myeloma cell lines by regulating miR-128-3p/WEE1 axis

FANG Yuan, LI Yi, WANG Wei

2024, 44(2):  210-218.  doi:10.16352/j.issn.1001-6325.2024.02.0210

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Objective To investigate the impacts of circular RNA RERE (circRERE) on proliferation, migration and invasion of human multiple myeloma (MM) cell lines by regulating microRNA (miR)-128-3p/serine/threonine protein kinase (WEE1) axis. Methods Normal plasma cells (nPCs) isolated from peripheral blood of healthy subjects and human MM cell lines(U266, RPMI-8226, NCI-H929, LP-1) were cultured. The expressions of circRERE and miR-128-3p were detected by RT-qPCR. The expression of WEE1 was detected by Western blot. After transfection, RPMI-8226 cells were divided into control group (without transfection), sh-circRERE group, sh-NC group, miR-128-3p inhibitor group, inhibitor-NC group, sh-circRERE+inhibitor-NC group and sh-circRERE+miR-128-3p inhibitor group. MTT assay and 5-ethynyl-2'-deoxyuridine (EdU) immunofluorescence staining microscopy were used to detect the proliferation. Migration and invasion were examined by scratch healing experiment as well as Transwell chamber assay. The targeting relationship between miR-128-3p and circRERE or WEE1 was verified though dual-luciferase reporter gene assay. Results Compared with nPCs, the expressions of circRERE and WEE1 in MM cells (RPMI-8226, U266, NCI-H929, LP-1) were increased, and the expression of miR-128-3p was decreased (P＜0.05). Compared with the control group, the proliferation rate, proportion of EdU positive cells, the healing rate of the scratch trauma and the number of invasion of the RPMI-8226 cells in sh-circRERE group were decreased(P＜0.05). However,miR-128-3p inhibitor group showed an opposite result and the difference was statistically significant (P＜0.05). miR-128-3p inhibitor significantly inhibited the effects of sh-circRERE on the proliferation, migration and invasion of RPMI-8226 cells (P＜0.05). Dual luciferase reporter gene experiments showed that circRERE/miR-128-3p and miR-128-3p/WEE1 had a targeting relationship. Conclusions CircRERE may promote proliferation, migration and invasion of RPMI-8226 cells, potentially with a mechanism of regulating miR-128-3p/WEE1 axis.

Omeprazole combined with different probiotics regulates intestinal microbiota to alleviate functional dyspepsia in children

HE Yun, XIAO Li, CAO Juan, LIU Zhigang, LUO Weiyao

2024, 44(2):  219-224.  doi:10.16352/j.issn.1001-6325.2024.02.0219

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Objective To explore the effect of omeprazole combined with different probiotics on regulating intestinal flora in reducing functional dyspepsia (FD) in children. Methods Two hundreds children with FD admitted to the Pediatric Department of Foshan Maternal and Child Health Hospital from January 2022 to February 2023 were selected as the study subjects. They were randomly divided into omeprazde(omep) group, groups of omeprazole +yeast(yeast group), +clostridium butyricum(clos group), and +bifidobacterium(bifi group) respectively. Results After treatment, serum level of IL-6, TNF-α, IL-1β, hs-CRP, VIP, SS, Enterobacter and Enterococcus in all groups significantly decreased as compared with the finding before treatment (P＜0.05). Those targets in the three combined treatment groups were significantly lower compared to the ome group; After treatment, the serum MOT level,bifidobacteria, and lactobacilli in each group were significantly increased (P＜0.05), and the results from three combined treatment groups demonstrated notably higher levels compared to the omep group(P＜0.05); The scores of symptoms in all groups showed a significant alleviation after the treatment(P＜0.05). Additionally, the three combined treatment groups exhibited significantly lower symptom scores than the group treated with omeprazole alone (P＜0.05). There was no difference in the incidence of adverse reactions during treatment among the groups. Conclusions Omeprazole combined with different probiotics have achieved good results in the treatment of FD in children.

Expression of leptin and its receptor correlates with hypoxia- inducible factor-1α in infants with cyanotic congenital heart disease

GUO Rong, ZHANG Sen, YUAN Jianhui, LI Xiaojue, ZHOU Shuhan, LI Shoujun, CHEN Yanyan

2024, 44(2):  225-230.  doi:10.16352/j.issn.1001-6325.2024.02.0225

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Objective To investigate the correlation between serum leptin level and body mass index (BMI) in infants with cyanosis congenital heart disease, and the relationship between leptin and Ob gene receptor (Ob-R) and hypoxia-inducible factor 1α (HIF-1α) in myocardium. Methods A total of 52 children under 6 months of age with congenital heart disease who underwent surgical treatment in the Department of Congenital Heart Surgery, Fuwai Hospital from January 2019 to October 2020 were included in this study. According to the arterial partial pressure of oxygen (PaO₂) of 90 mmHg, they were divided into cyanotic group (n=30) and acyanotic group (n=22). Their height and weight were collected to calculate BMI. The serum leptin level was measured by ELISA. The expressions of HIF-1α and Ob-R in myocardial tissue were detected by RT-PCR and Western blot. In animal model, SD rats were divided into normoxia group and hypoxia intervention group, which were subjected to continuous hypoxia (10% O₂) for 4 weeks. The hypoxia intervention group received intraperitoneal injection of HIF-1α inhibitor digoxin (2 mg/kg) daily from the 14 th to 21st day of hypoxia, respectively. The body weight of rats was recorded, and the expressions of HIF-1α and Ob-R were detected by RT-qPCR and Western blot. Results Compared with the acyanosis group, the cyanosis group had a significantly lower BMI (P＜0.05) and a lower leptin/BMI ratio (leptin /BMI) (P＜0.05). Spearman correlation analysis confirmed that serum leptin in the circulatory system was positively correlated with BMI (P＜0.05). In the cyanosis group, the expression of Ob-R increased with the upregulation of HIF-1α,showing a positive correlation. In animal model, with the down-regulation of HIF-1α expression in digoxin injection, the Ob-R level was significantly lower than that in the control group(P＜0.05), the trend of weight loss was significantly inhibited(P＜0.05). The right ventricular hypertrophy index was significantly lower than that in the control group (P＜0.05). Conclusions HIF-1α regulates the expression of Ob-R in myocardial tissue, and the mechanism of its association with leptin and Ob-R may help to find new therapeutic target for improving the prognosis of infants with congenital heart disease.

Improving circadian rhythm disturbance reduces myocardial ischemia-reperfusion injury in diabetic rats

QIN Xiaoying, LIU Hui, HAN Chongfang, HE Jiandong

2024, 44(2):  231-234.  doi:10.16352/j.issn.1001-6325.2024.02.0231

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Objective To investigate whether discontinuous sleep supplementation can reduce myocardial ischemia-reperfusion injury in diabetic rats aggravated by circadian rhythm disorder. Methods The rats were injected intra-peritoneal with 1% streptozotocin(STZ) 30 mg/kg combined with high-fat and high-glucose diet to replicate diabetic model. Forty diabetic rats were randomly divided into four groups with 10 in each: sham surgery group(Sham group),ischemia-reperfusion group(I/R group), in which the left anterior descending coronary artery(LDA) was ligated for thirty minutes and reperfusion for 2 h,circadian rhythm disorder group(Crd group,24 h daily light and food),discontinuous sleep supplementation group(Dss group,every 3 hours of illumination and 1.5 hours break at night). We analyzed the myocardial infarct size (by 2,3,5-triphenyltetrazolium chloride staining),determined serum creatine kinase-myoglobin(CK-MB) activity and cardiac troponin Ⅰ(cTnⅠ) concentrations; the expression level of BMAL1 and REV-ERBα was determined by Western blot. Results Compared to the sham group, the I/R group showed a significantly increased in myocardial infarct size, serum CK-MB activity and cTnⅠ concentration. The expression of the myocardial biological clock gene BMAL1 was down-regulated, while the expression of REV-ERBα was up-regulated(P＜0.05).Compared to the I/R group, the Crd group showed a significantly increase in myocardial infarct size, serum CK-MB activity and cTnⅠ concentration. The expression of the myocardial biological clock gene BMAL1 was down-regulated, while the expression of REV-ERBα was up-regulated (P＜0.05).Compared to the Crd group, Dss group showed a significantly decrease in the myocardial infarct size, serum cTn concentration and CK-MB activity.Furthermore,there was an increased protein expression of BMAL1 and a decrease of REV-ERBα(P＜0.05). Conclusions Discontinuous sleep supplementation can reduce myocardial ischemia-reperfusion injury in diabetic rats aggravated by circadian rhythm disorder.

Preparation of Lir@BSA-PMF nanoparticles and verification of their cell functions

HUANG Qingyu, CHEN Qiying, SUN Shengjia, WU Bangwei, LIN Shan, Alimujiang·MAIMAITIJIANG

2024, 44(2):  235-241.  doi:10.16352/j.issn.1001-6325.2024.02.0235

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Objective To synthesize bovine serum albumin(BSA)-loaded liraqlutide (Lir)-nanoparticles coated with platelet membrane fragments (PMF) using a“bottom-up” nano-engineering chemistry technique, and to evaluate their cyto-compatibility and potential function of anti-oxidative stress. Methods PMF was extracted as reported previously. Lir@BSA nanoparticles were prepared by self-assembly method. PMF was coated on the surface of Lir@BSA nanoparticles by co-extrusion to prepare Lir@BSA-PMF. The physical and chemical properties of Lir@BSA-PMF particles were characterized as particle size, Zeta potential, transmission electron microscopy and particle size stability. The encapsulation efficiency, loading efficiency and cumulative release efficiency of liraglutide were calculated by enzyme-linked immunosorbent assay. Further, SDS-PAGE was used to analyze whether there was a similar membrane protein distribution of platelet membrane on Lir@BSA-PMF bionicnanocarrier.CCK-8 assay was used to verify the biocompatibility of the materials. Reactive oxygen species (ROS) experiment was used to explore the effect of Lir@BSA-PMF on cell oxidative damage. The uptake of cells on Lir@BSA-PMF bionic nano capsules was verified by cell phagocytosis experiment. Results Lir@BSA-PMF nanoparticles had a stable particle size of 25 nm with a spherical morphology, and a Zeta potential value of -25.5 mV. The encapsulation efficiency, loading efficiency and cumulative release efficiency of liraglutide were 85.56%, 7.96% and 77.06%, respectively. SDS-PAGE analysis showed that the Lir@BSA-PMF bio-mimetic nano capsules retained the similar membrane protein distribution as platelet membrane. CCK-8 assay verified that the nanomaterials were non-cytotoxic. ROS results showed that Lir@BSA-PMF nanomaterials had obvious antioxidant properties. The results of cell phagocytosis showed that the cells had a good phagocytosis effect on Lir@BSA-PMF nanoparticles. Conclusions The nanoparticles Lir@BSA-PMF are successfully synthesized and have no effects on cells viability in vitro. The particles are taken up by cells and show a significant function of antioxidant damage.

Clinical Sciences

Improvement of early exercise combined with electrical stimulation of neuromuscular system on ICU acquired weakness in patients with severe pneumonia

LU Yunxia, FENG Yue, JIANG Jinxia, YANG Shuai

2024, 44(2):  242-246.  doi:10.16352/j.issn.1001-6325.2024.02.0242

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Objective To investigate the effect of early activity combined with electrical stimulation of neuromuscular system in improving intensive care unit (ICU)-acquired weakness (ICU-AW) patients with severe pneumonia. Methods A total of 150 patients with ICU-AW caused by severe pneumonia admitted to emergency intensive care unit(EICU) in a tertiary hospital in Shanghai were enrolled as the study subjects, and randomly divided into control group (75 cases) and combined group (75 cases). The control group took early exercise, and the combination group was given early exercise plus electrical stimulation of neuromuscular system. The recovery from mechanical ventilation (ICU length of stay, duration of mechanical ventilation, weaning extubation rate, ICU rollout rate), lung function [forced vital capacity (FVC), forced expiratory volume in the 1 s (FEV1)/FVC, massive inspiratory pressure (MIP)],muscle strength[Medical Research Council (MRC) score], disease severity[acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ)], and complications were compared between the two groups. Results The ICU length of stay and mechanical ventilation time in the combined group were significantly lower than those in the control group (P＜0.05). After intervention, FVC, FEV1/FVC, MIP and MRC scores in the two groups were significantly increased(P＜0.05), and compared with the control group, the combined group was significantly increased(P＜0.05). After intervention, APACHE Ⅱ score was significantly reduced in the two groups(P＜0.05), and compared with the control group, APACHE Ⅱ score in combined group were significantly reduced (P＜0.05). The complication rate in the combined group (9.33%) was significantly lower than that of control group (24.00%) (P＜0.05). Conclusions Early exercise combined with neuromuscular electrical stimulation in patients with severe pneumonia ICU-AW can effectively promote recovery of patients because of improved lung function. This therapy is proved to be be safe and effective.

Efficacy of cinobufacini capsule combined with docetaxel plus cisplatin chemotherapy for the treatment of non-small cell lung cancer

LI Juan, LI Xiaofeng, XU Shengzhi, TANG Kai

2024, 44(2):  247-251.  doi:10.16352/j.issn.1001-6325.2024.02.0247

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Objective To investigate the impacts of cinobufotalin combined with docetaxel+cisplatin chemotherapy on the disease control and the level of vascular endothelial growth factor (VEGF) in serum of patients with non-small cell lung cancer. Methods Totally 70 patients with non-small cell lung cancer admitted to Zhejiang Veteran Hospital from March 2019 to March 2022 were included. They were randomly divided into combination group (cinobufotalin combined with docetaxel+cisplatin,n=35) and control group (docetaxel+cisplatin chemotherapy alone,n=35).The disease control rates of two groups, serum VEGF level, carcinoembryonic antigen (CEA), cytokeratin 19 (CK19) and carbohydrate antigen 125 (CA125) were measured using ELISA; life quality of patients was evaluated with the KPS score; The adverse events between two groups were compared. Results The disease control rate in the combination group (33/35, 94.29%) was higher than that that of control group (25/35, 71.43%,) (P＜0.05). After treatment, the level of VEGF in both groups of patients decreased, and the KPS score increased (P＜0.05) especially in the combination group (P＜0.05). After treatment, the level of serum CEA, CA125, and CK19 of control group and combination group were obviously lower than those before treatment(P＜0.05), while those in the combination group were even lower (P＜0.05). The incidence of adverse events in the combination group (5/35, 14.29%) was lower than that in the control group (13/35, 37.14%) (P＜0.05). Conclusions Cinobufotalin combined with docetaxel+cisplatin chemotherapy as a potential new chemotherapy significantly reduces the level of VEGF and tumor biomarkers in serum factor, improves the life quality of patients.The combined therapy is proved tobe safe.

Low-frequency electrical stimulation relieves pain and promotes gastrointestinal function recovery after gynecological laparotomy

YANG Yajing, ZHU Weipei, ZHOU Liulin

2024, 44(2):  252-255.  doi:10.16352/j.issn.1001-6325.2024.02.0252

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Objective To observe the effect of low-frequency electrical stimulation on the rehabilitation of patients after gynecological abdominal surgery. Methods Sixty-three patients who underwent open surgery in gynecology department of Taixing Clinical College of Bengbu Medical College from June 2021 to July 2022 were selected. The patients were randomly divided into control group (31 cases) and a low-frequency electrical stimulation group (32 cases). The low-frequency electrical stimulation group was subjected to stimulation within the patient's tolerable range once a day for 30 minutes each time, and the intensity of each stimulation was adjusted based on clinical situation. The control group selected the same acupoints and pasted electrodes, connected to the treatment device but no electrical stimulation. The electrode strip was removed after 30 minutes,then record the postoperative Visual Analog Scale (VAS) score as well as the time from the end of the surgery to the first discharge and defecation. Results The VAS score at 48 hours after surgery showed a low degree of pain in the low-frequency electrical stimulation group (3.6±1.2) compared to that in control group (4.5±1.4); After 72 hours of surgery, the VAS score was lower in the low-frequency electrical stimulation group (1.7±0.9) compared to the control group (3.3±1.4), indicating a lower degree of pain. The first exhaust time (26.9±6.7)h vs. (35.5±13.0)h was shorter in the low-frequency electrical stimulation group; The first bowel movement time (49.0±5.4)h vs. (64.4±13.8)h was shorter in the low-frequency electrical stimulation group compared to the control group. Conclusions Low frequency electro-physiological stimulation can alleviate post-operative pain and shorten exhaust and defecation time in patients undergoing gynecological open surgery.

Mini Reviews

Advances in the role of lysophosphatidyl-choline acyltransferase 1 in tumors

CHEN Xue, LUO Tian, WEI Chaojun

2024, 44(2):  256-259.  doi:10.16352/j.issn.1001-6325.2024.02.0256

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Lysophosphatidylcholine acyltransferase 1(LPCAT1), a key metabolic enzyme in the phosphatidyl-choline metabolism pathway, mediates phosphatidylcholine synthesis through deacylation-reacylation, which leads to alterations in the phospholipids composition of cell membranes and the remodelling of the cellular cytoskeleton. Lysophosphatidylcholine acyltransferase 1 has been shown to be highly expressed in gastric, breast and colorectal cancer, and can accelerate the alteration of the phospholipids composition of tumor cell membranes or interact with tumor driver genes, such as epidermal growth factor receptor and cell cycle-related genes. It might promote tumor development by affecting tumor cell proliferation, migration, invasion and resistance to chemotherapy.

Research progress on the role of ferroptosis in the pathogenesis of non-alcoholic fatty liver disease

XIAO Yanxin, LIU Yan, XU Lingling, ZHOU Yaru

2024, 44(2):  260-264.  doi:10.16352/j.issn.1001-6325.2024.02.0260

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Ferroptosis is a new type of cell death proposed in recent years, and its main characteristics are iron overload and lipid peroxidation. Ferroptosis is involved in the occurrence and development of non-alcoholic fatty liver disease(NAFLD). Iron overload can generate a large amount of reactive oxygen species through the Fenton reaction. Under the action of lipoxygenase, the unsaturated fatty acids on the liver cell membrane undergo lipid peroxidation, which induces liver cell death and leads to the occurrence of non-alcoholic fatty liver disease/non-alcoholic steatohepatitis. Blocking ferroptosis may provide one of the therapeutic strategies to protect liver cells.

Early imaging and circulating marker features of gallbladder carcinoma

ZHAO Yongzhong, XUE Yi, LI Liqun, HOU Mengsen, YANG Xiaojun

2024, 44(2):  265-269.  doi:10.16352/j.issn.1001-6325.2024.02.0265

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Gallbladder carcinoma (GBC) is one of the most common malignant tumors in the biliary system, which is difficult to diagnose in the early stage due to its high degree of malignancy, invasiveness and lack of specific clinical manifestations. In this paper, we summarize ultrasound, CT and other imaging manifestations in the early stage of GBC, and describe the role of protein markers and microRNA marker as biomarkers in the diagnosis of early GBC. The enhanced understanding of the relevant features might help to improve the accuracy of the diagnosis of early gallbladder carcinoma.

Medical Education

Investigation of continuing education requirements on the diagnosis and management of pain in non-anesthetic residents

WANG Jin, LIU Hongju, SHEN Le

2024, 44(2):  270-275.  doi:10.16352/j.issn.1001-6325.2024.02.0270

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Objective To investigate the current status and training requirements of pain medicine among non-anesthetic residents rotating in the Department of Anesthesiology at a tertiary hospital in Beijing. Methods A self-designed questionnaire of “pain medicine education requirements of rotating residents” was administered to each resident on their first day of rotation, and voluntary responses were collected. Results A total of 108 questionnaires were collected (87.1% response rate), comprising 45 from surgical residents, 42 from internal medicine residents, and 21 from emergency and intensive care residents. The results showed that surgical residents encountered a higher prevalence of surgery-related pain as compared to internal medicine residents or emergency and intensive care residents(P＜0.001). Internal medicine residents experienced a greater incidence of inflammatory pain than surgical residents(P＜0.05) or emergency and intensive care residents(P＜0.01). All residents expressed continuing education requirements on pain management. Regarding training modalities, surgical residents favored medical course (86.7%); internal medicine residents preferred outpatient clinic rotation (88.1%); and emergency and intensive care residents preferred interventional treatment rotations (90.5%). Conclusions There are a wide need for continuing medical education on pain management. The training should be tailored diversely based on the specific clinical needs of each department to improve the overall quality of continuing medical education and accelerate the development of integrative pain management.

Exploration and practice of Enhanced Recovery after Surgery and Perioperative Management course in postgraduate teaching

ZHU Qianmei, LIU Zijia, TAN Gang, SHEN Le, HUANG Yuguang

2024, 44(2):  276-280.  doi:10.16352/j.issn.1001-6325.2024.02.0276

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Objective To explore the practice and significance of the new course of “ Enhanced Recovery after Surgery (ERAS) and Perioperative Management” for graduate students under multidisciplinary cooperation. Methods The Department of Anesthesiology collaborated with the Department of Clinical Nutrition, Department of Geriatrics and six related surgical departments to develop a course of 30 credit hours on “ ERAS and Perioperative Management” in Peking Union Medical College Hospital. Researchers analyzed the teaching effectiveness of the course by collecting survey questionnaires and evaluating scheme report of ERAS case. Results Researchers found that ten graduates joined this course and they believed that learning improved their understanding of ERAS related knowledge, such as preoperative nutrition and functional state optimization, intraoperative volume and temperature management, prevention of postoperative nausea and vomiting, and perioperative pain management. Students had high satisfaction with the course and believed that it would help improve their clinical literacy. Conclusions The new course of “ERAS and Perioperative Management” for graduates may support capacity building in terms of clinical logic and thinking about ERAS and promotion of skills for perioperative management. Our experience of graduates'training with “ Enhanced Recovery after Surgery and Perioperative Management” course can be shared by other trainers of health institution of China.