Tianjin Medical Journal

MiR-338-5p can promote the proliferation, migration and invasion of bladder cancer cells by targeting TSHZ3

LIU Hongwei, ZHU Yi, XIANG Lingbao, XIONG Hong, CHEN Ruiqi

2023, 51 (10): 1025-1031. doi: 10.11958/20221910

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Objective To investigate the mechanism of miR-338-5p regulating the proliferation, migration and invasion of bladder cancer cells by targeting TSHZ3. Methods The expression of miR-338-5p in 33 samples of bladder cancer tissue and paracancerous tissue was detected by qRT-PCR. Bladder cancer T24 and UM-UC-3 cells were transfected with mimics-NC, miR-338-5p mimics, inhibitor-NC and miR-338-5p inhibitor, and the transfection efficiency was detected by qRT-PCR. CCK-8 assay was used to detect the effect of overexpression or knockdown of miR-338-5p on the proliferation of bladder cancer cells. Transwell assay was used to detect the effect of miR-338-5p on migration and invasion of bladder cancer cells. The potential target genes of miR-338-5p were predicted by TargetScan, miRDB and Targetminer databases, and TSHZ3 was chosen for the target gene. The targeting relationship between miR-338-5p and TSHZ3 was verified by dual-luciferase reporter gene assay and Western blot assay. miR-338-5p mimics and TSHZ3 overexpression plasmids were co-transfected in bladder cancer cells, and the proliferation, migration and invasion of cells were detected by CCK-8 and Transwell assays. Western blot assay was used to detect the overexpression of miR-338-5p or TSHZ3 against Wnt/ β-catenin signaling pathway. Results miR-338-5p was significantly upregulated in bladder cancer (P<0.05). Overexpression of miR-338-5p increased the proliferation, migration and invasion of bladder cancer cells (P<0.05), while transfection of miR-338-5p inhibitor decreased the proliferation, migration and invasion of bladder cancer cells (P<0.05). Bioinformatics analysis and the dual-luciferase reporter gene assay showed that there was a targeting relationship between miR-338-5p and TSHZ3. The rescue experiment showed that co-transfection of miR-338-5p mimics and TSHZ3 overexpression plasmid partially eliminated the promoting effect of overexpression of miR-338-5p on the proliferation, migration and invasion of bladder cancer cells (P<0.05). Western blot results showed that overexpression of miR-338-5p significantly decreased the expression level of TSHZ3 protein and increased Wnt3a and β-catenin protein expression levels (P<0.05). In addition, overexpression of TSHZ3 reduced Wnt3a and β-catenin protein expression levels (P<0.05). Conclusion miR-338-5p is upregulated in bladder cancer, and promotes the proliferation, migration and invasion of bladder cancer by targeting TSHZ3 expression. The mechanism may be related to the activation of Wnt/β-catenin signaling pathway.

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Effects of Mesothelin on proliferation, apoptosis, migration and invasion of breast cancer MDA-MB-231 cells

LI Yuxiang, LI Ge, GAO Zhenfang, HOU Chunmei, HAN Gencheng, SUN Huiyan, WANG Lisheng

2023, 51 (10): 1032-1039. doi: 10.11958/20230452

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Objective To investigate the effect of Mesothelin on proliferation, apoptosis, migration and invasion of human triple-negative breast cancer cells (TNBC). Methods Immunohistochemical staining was used to detect the expression of Mesothelin in TNBC tissue, paracancer tissue and breast cancer cell lines. TNBC cell line MD-MB-231 cells were transfected with the overexpressed Mesothelin lentivirus plasmid (the mesothelin group) and the control Scramble plasmid (the Scramble group). MD-MB-231 cells with no infected lentivirus were used as the control group. Flow cytometry was used to detect the efficiency of virus infection. The expression of Mesothelin was detected by qPCR, flow cytometry and Western blot assay. Cell proliferation was detected by CCK-8. Annexin V-APC/PI was used to detect apoptosis. The expression of anti-B lymphoblastoma-2 (Bcl-2), anti-Bcl-2 associated X protein (Bax), E-cadherin, N-cadherin and Vimentin were detected by Western blot assay. Cell migration ability was detected by scratch test. The invasive ability of cells was detected by Transwell assay. Results The expression of Mesothelin was significantly higher in TNBC tumor tissue than that in paracancer tissue (P<0.05), and Mesothelin was expressed in breast cancer cell line. Flow cytometry showed that the efficiency of lentivirus infection was more than 90%. Compared with the Scramble group, the expression level of Mesothelin mRNA was increased in the Mesothelin group, and the cell proliferation rate was increased, while the cell apoptosis rate was decreased. The expression of Bax and E-cadherin protein decreased, the expression of Bcl-2, N-cadherin and Vimentin protein increased, and the migration and invasion ability of cells were significantly enhanced (P<0.05). Conclusion Mesothelin can promote the proliferation, inhibit the apoptosis and increase the ability of migration and invasion of human triple-negative breast cancer MDA-MB-231 cells.

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Impacts of Casticin on biological behavior of gastric cancer cells by regulating miR-378/PRRX1 axis

ZHANG Cong, FENG Hua, HUANG Xiaolong, WANG Xuan

2023, 51 (10): 1040-1047. doi: 10.11958/20222053

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Objective To explore effects of Casticin (Cas) on the proliferation, invasion, migration and microRNA-378 (miR-378)/paired related homeobox 1 (PRRX1) axis of gastric cancer (GC) SGC-7901 cells. Methods SGC-7901 cells were treated with Cas at different concentrations (0, 5, 10, 20, 40 and 80 μmol/L) for 48 h, and the cell viability of SGC-7901 was detected by tetrazole salt (MTT) method. SGC-7901 cells at logarithmic growth stage were divided into 7 groups: the control group, the Cas low concentration group (10 μmol/L), the Cas medium concentration group (20 μmol/L), the Cas high concentration group (40 μmol/L), the Cas high concentration+miR-378 small interfering RNA (siRNA) group (40 μmol/L Cas+miR-378 siRNA), the Cas high concentration+PRRX1 group (40 μmol/L Cas+PRRX1 overexpressed plasmid) and the Cas high concentration+miR-378 siRNA+PRRX1 group (40 μmol/L Cas+miR-378 siRNA+PRRX1 overexpression plasmid). Cell activity, colony formation number, apoptosis rate, invasion and migration ability of SGC-7901 cells were determined by MTT assay, colony formation assay, flow cytometry, Transwell chamber assay and scratch assay, respectively. Expression levels of miR-378 and PRRX1 protein in SGC-7901 cells were detected by fluorescence quantitative PCR and Western blot assay. Double luciferase assay verified the targeting relationship between miR-378 and PRRX1. Xenografted SGC-7901 cells were xenografted into nude mice to verify the anticancer activity of Cas and its effect on the expression of miR-378. Results The cell viability of SGC-7901 cells in Cas 10, 20, 40 and 80 μmol/L groups decreased in a concentration-dependent manner (P<0.05), and 10, 20 and 40 μmol/L Cas were selected as the follow-up study concentration. Compared with the control group, the cell viability, colony formation number, invasion cell number, scratch healing rate and PRRX1 protein expression level of SGC-7901 cells in the Cas low, medium and high concentration groups were decreased successively (P<0.05), while the apoptosis rate and the expression level of miR-378 were increased successively (P<0.05). Compared with Cas high concentration group, the cell viability, colony formation number, invasion cell number, scratch healing rate and PRRX1 protein expression levels were increased in the Cas high concentration+miR-378 siRNA group and the Cas high concentration+PRRX1 group, and the apoptosis rate was decreased (P<0.05). The expression level of miR-378 in SGC-7901 cells was decreased in the Cas high concentration+miR-378 siRNA group (P<0.05). Compared with the Cas high concentration+miR-378 siRNA group, the expression level of miR-378 in SGC-7901 cells was increased in the Cas high concentration+PRRX1 group (P<0.05). Compared with the Cas high concentration+miR-378 siRNA group and the Cas high concentration+PRRX1 group, the cell viability, colony formation number, invasion cell number, scratch healing rate and PRRX1 protein expression levels were increased in the Cas high concentration+miR-378 siRNA+PRRX1 group (P<0.05), and the apoptosis rate was decreased (P<0.05). In SGC-7901 cells, miR-378 could target the expression of PRRX1. Cas inhibited the growth of transplanted tumor and promoted the expression of miR-378 in nude mice (P<0.05). Conclusion Cas may inhibit the proliferation, invasion and migration of SGC-7901 cells by regulating the miR-378/PRRX1 axis.

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Influence of circACTR2 on high glucose-induced trophoblast cell injury by regulating miR-23a-3p/TBL1X axis

HUANG Chunyan, XIANG Mi, FEI Zhiyi, GAO Qin

2023, 51 (10): 1048-1053. doi: 10.11958/20230156

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Objective To investigate the influence of circular RNA actin related protein 2 (circACTR2) on high glucose-induced trophoblast cell injury by regulating miR-23a-3p/transducin β1X-linked protein (TBL1X) axis. Methods Human chorionic trophoblast cells HTR-8/Svneo were grouped into the NG group (5.5 mmol/L glucose), the HG group (25 mmol/L glucose), the si-NC group (25 mmol/L glucose+transfected with si-NC), the si-circACTR2 group (25 mmol/L glucose+transfected with si-circACTR2), the si-circACTR2+inhibitor-NC group (25 mmol/L glucose+co-transfected with si-circACTR2 and inhibitor-NC) and si-circACTR2+miR-23a-3p inhibitor group (25 mmol/L glucose+co-transfected with si-circACTR2 and miR-23a-3p inhibitor). Real-time quantitative PCR (qPCR) was performed to measure expression levels of circACTR2 and miR-23a-3p in cells. Cell proliferation was detected by CCK-8 assay. Cell apoptosis was detected by flow cytometry. Scratch assay was performed to detect cell migration. Enzyme-linked immunosorbent assay was applied to detect malondialdehyde (MDA) level, lactate dehydrogenase (LDH) and superoxide dismutase (SOD) activities. Western blot assay was used to detect expressions of TBL1X, proliferating cell nuclear antigen (PCNA), matrix metalloproteinase-2 (MMP-2), MMP-9 and cysteine protease-3 (caspase-3) in cells. Dual-luciferase reporter gene experiment was performed to verify the targeting relationship between circACTR2, TBL1X and miR-23a-3p respectively. Results Compared with the NG group, the HTR-8/Svneo cell expression of miR-23a-3p, proliferative ability, scratch healing rate, expression of PCNA, MMP-2, MMP-9 and SOD activity were reduced in the HG group, and the expression of circACTR2, TBL1X, caxpase-3, MDA content, LDH activity and apoptosis rate were increased (P<0.05). Compared with the HG group and the si-NC group, the miR-23a-3p expression, proliferative ability, scratch healing rate, expression of PCNA, MMP-2 and MMP-9, and SOD activity in HTR-8/Svneo cells were increased in the si-circACTR2 group, and the expression of circACTR2, TBL1X and caxpase-3, MDA content, LDH activity and apoptosis rate were decreased (P<0.05). On the basis of knocking down circACTR2, the down-regulation of miR-23a-3p significantly attenuated the promotion of circACTR2 knockdown on the proliferation and migration of high glucose-induced HTR-8/Svneo cells, and enhanced the ability of apoptosis and oxidative stress. The results of dual-luciferase reporter gene assay showed that circACTR2 targeted to negatively regulate the expression of miR-23a-3p, and miR-23a-3p targeted to negatively regulate the expression of TBL1X. Conclusion Knocking down circACTR2 can inhibit high glucose-induced trophoblast cell injury and exert a protective effect by regulating miR-23a-3p/TBL1X axis.

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The protective effect and mechanism of melatonin on myocardial ischemic reperfusion injury

ZHENG Junyi, LI Xiaofeng, GUO Xukun

2023, 51 (10): 1054-1058. doi: 10.11958/20231005

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Objective To investigate the protective effect of melatonin on myocardial ischemia reperfusion injury(IRI) by regulating Akt/FOXO4/BIM signaling pathway and its mechanism. Methods Myocardial IRI model was established, and cells were divided into the control group (CTRL group), the ischemia reperfusion group (IRI group), the melatonin treatment group (IRI+MEL group) and the Akt inhibitor group (IRI+MEL+MK2206 group). MTT method was used to detect the cell survival rate. The level of released LDH was measured with microplate reader. Flow cytometry was used to detect cell apoptosis. PCR method was used to detect the inflammatory gene levels of interleukin 6 (IL-6), tumor necrosis factor α (TNF-α) and monocyte chemotactic protein-1 (MCP-1). Western blot assay was used to detect the protein expression changes of p-Akt, p-FOXO4 and BIM. Results Compared with the CTRL group, the cell survival rate decreased and LDH content increased, the early stage and total apoptosis rates were increased, expression levels of IL-6, TNF-α and MCP-1 genes were increased, p-Akt protein expression decreased, and p-FOXO4 and BIM protein expression levels increased in the IRI group (P<0.05). Compared with the IRI group, melatonin therapy increased the cell survival rate and p-Akt protein expression, decreased LDH content, cell apoptosis, IL-6, TNF-α, MCP-1 gene level and p-FOXO4 and BIM protein expression (P<0.05). Compared with the IRI+MEL group, MK2206 partially reversed the protective effect of melatonin on IRI cardiomyocytes (P<0.05). Conclusion Melatonin has protective effect on ischemia reperfusion injured myocardial cells, and its mechanism may be related to the regulation of Akt/FOXO4/BIM signaling pathway to reduce apoptosis and cell inflammation.

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Micheliolide ameliorates renal lesion of unilateral ureteral obstruction mice by inhibiting NF-κB/NLRP3 axis

LEI Xianghong, YAN Wenjun, XIONG Meimei, LONG Haibo, CHEN Sijia

2023, 51 (10): 1059-1064. doi: 10.11958/20230150

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Objective To investigate the effect and mechanism of micheliolide on renal lesions of unilateral ureteral obstruction (UUO) mice. Methods Male C57BL/6J mice were randomly divided into the sham group, the UUO (model) group and the UUO+micheliolide (administration) group. Mice in the UUO+ micheliolide group were given micheliolide by gavage at 25 mg/kg body weight after the establishment of the animal model. Renal tissue samples were collected 8 days after operation, and the renal tissue was stained with HE, Masson and TUNEL staining. The protein expressions of nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3), caspase-1, interleukin (IL)-1β, tumor necrosis factor (TNF)-α and (phosphorylation)- nuclear transcription factor (NF)-κB p65 were detected by immunohistochemistry and Western blot assay. Results HE staining showed that there were no pathological changes in renal tissue in the sham group. Mice in the UUO group had progressive renal tubule dilation and interstitial edema with a small amount of inflammatory cell infiltration. The renal lesions were significantly improved in the UUO+MCL group compared with those of the UUO group. Masson staining showed that compared with the sham group, the collagen volume fraction (CVF) was significantly increased in the UUO group, while CVF was significantly reduced in the UUO+MCL group than that in the UUO group. TUNEL staining showed that compared with the sham group, the apoptosis rate was increased in the UUO group, and the apoptosis rate was significantly decreased in the UUO+MCL group than that of the UUO group. Western blot assay and immunohistochemistry result showed that the expression levels of NLRP3, caspase-1, IL-1β, TNF-α and p-NF-κB p65 were increased in kidney tissue of the UUO group compared with those of the sham group. Compared with the UUO group, NLRP3, caspase-1, IL-1β, TNF-α and p-NF-κB p65 were significantly decreased in the UUO+MCL group. Conclusion The expression of NLRP3 inflammasome and related inflammatory factors in renal tissue of mice with unilateral ureteral obstruction are increased. Micheliolide can reduce the inflammatory response of renal tubule interstitium by inhibiting NF-κB pathway and NLRP3 inflammasome activation, thereby improving renal fibrosis.

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Naa20 alleviated myocardial injury in sepsis by inhibiting AIM2 expression

LIU Chang, FAN Caihong, LIU Jia, LIU Yashan, ZHANG Shiqi, LIU Shiming, SHEN Yanna

2023, 51 (10): 1065-1070. doi: 10.11958/20230338

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Objective To explore the regulatory effect and mechanism of Naa20 on lipopolysaccharide (LPS) induced myocardial injury in sepsis. Methods Twelve male C57BL/6J mice were divided into the control group and the myocardial injury model group (n=6 in each group). Mice of the myocardial injury model group were treated with a single intravenous injection of LPS, while mice of the control group were injected with the same dose of PBS. Cardiac function of mice in each group was evaluated by cardiac Doppler ultrasonography 12 h after LPS injection. The serum content of CK-MB was detected by enzyme-linked immunosorbent assay (ELISA). Hematoxylin-eosin staining (HE) was used to evaluate myocardial injury in mice. Cardiomyocytes from SD rats were isolated and cultured in vitro and divided into the control group and the LPS group. Mitochondrial membrane potential and reactive oxygen species (ROS) release in cardiomyocytes were detected by fluorescent probe. The protein expression levels of AIM2, GSDMD, activated GSDMD (GSDMD-N), IL-1β, activated IL-1β (p17) and Naa20 were detected by Western blot assay. The content of IL-1β in supernatant was determined by ELISA. The expression level of Naa20 in cardiomyocytes was detected by quantitative real-time PCR (qPCR). H9c2 cells were randomly divided into the control group, the Naa20 group, the LPS group and the Naa20+LPS group. Methyl thiazolyl tetrazolium (MTT) assay and lactate dehydrogenase (LDH) assay were used to detect the viability of H9c2 cells. The interaction between Naa20 and AIM2 was detected by immunoprecipitation. Results Compared with the control group, the left ventricular ejection fraction (LVEF) and left ventricular fractional shortening (FS) were decreased in the myocardial injury model group (P<0.01), the serum content of CK-MB was increased (P<0.01), and HE staining showed pathological changes in heart of mice. In vitro, compared with the control group, mitochondrial membrane potential was decreased, while ROS production, expressions of AIM2, GSDMD, GSDMD-N, IL-1β, p17 and Naa20 and the content of IL-1β in supernatant were significantly increased in the LPS group (P<0.05). Results of immunocoprecipitation confirmed that Naa20 interacted with AIM2. Compared with the LPS group, the viability of H9c2 cells and the acetylation of AIM2 were increased, while the protein expression levels of AIM2, GSDMD, GSDMD-N and the content of IL-1β in supernatant were decreased in the Naa20+LPS group (P<0.05). Conclusion Naa20 can regulate AIM2-mediacted pyroptosis by inhibiting AIM2 expression, thereby alleviating sepsis-induced myocardial injury.

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Protective effect and mechanism of inosine on acute lung injury induced by lipopolysaccharide in rats

XU Nongyu, ZHONG Jiangshan, LI Duo

2023, 51 (10): 1071-1075. doi: 10.11958/20230059

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Objective To investigate the lung protective effect of inosine on acute lung injury (ALI) rats and its associated mechanism. Methods Twenty SD rats were randomly divided into 4 group: the control (NC) group, the model (LPS) group, the inosine low-dose intervention (IN-L) group and the inosine high-dose intervention (IN-H) group. Rats challenged with intratracheal lipopolysaccharide (LPS) were treated after 1, 6, and 12 hours with 100 mg/kg inosine (IN-L group), 200 mg/kg inosine (IN-H group) and normal saline (LPS group). The NC group received equal volume normal saline during the modeling and intervention phases. After 24 hours of the induction of ALI, the arterial blood was used to detect partial pressure of oxygen (PaO₂), the lung tissue sample was collected for the wet/dry weight (W/D) value and pathological examination. Serum and bronchoalveolar lavage fluid (BALF) were obtained to measure polyadp ADP-ribose polymerase (PARP)-1, inducible nitric oxides (iNOS), tumor necrosis factor (TNF)-α and interleukin (IL)-1β. Lung tissue homogenate was prepared to determine the toll-like receptor 4 (TLR4), myeloid differentiation primary response 88 (MYD88) and nuclear factor kappa-B (NF-κB) proteins. Results LPS induced lung tissue injury in rats, resulting in severe hypoxemia, increased levels of PARP-1 and iNOS, promoted the secretion of inflammatory factors TNF-α and IL-1β, and up-regulated expression levels of TLR4, MYD88, NF-κB proteins in lung tissue (P<0.05). Inosine intervention down-regulated expression levels of TLR4, MYD88 and NF-κB in lung tissue, decreased the production of PARP-1, iNOS, TNF-α and IL-1β, alleviated the degree of lung tissue injury and improved hypoxemia (P<0.05). The above changes were more obvious in the IN-H group than those in the IN-L group (all P<0.05). Conclusion Inosine inhibits LPS-induced lung inflammation and reduces inflammatory cytokines, which may be related in part to the inosine down-regulation of TLR4/MYD88/NF-κB signaling pathway, the inhibition of PARP-1 activity and the reduction of NF-κB nuclear translocation.

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Effects of mild hypothermia treatment on the immune and neurological function in a swine model of extracorporeal cardiopulmonary resuscitation after cardiac arrest

WANG Jinxiang, XU Guowu, WANG Tailan, WANG Le, JIN Heng, CHAI Yanfen

2023, 51 (10): 1076-1079. doi: 10.11958/20230333

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Objective To investigate the effect of mild hypothermia treatment (MHT) on levels of immune and neurological function indicators in a swine model of extracorporeal cardiopulmonary resuscitation after cardiac arrest (CA). Methods Eighteen Bama miniature pigs were randomly divided into the sham group, the normal temperature (NT) group and the mild hypothermia (MHT) group with 6 pigs in each group. CA model was established by right ventricular fibrillation in the NT group and the MHT group. Then cardiopulmonary resuscitationt and temperature management were followed. The target temperature of the NT group was 37 ℃, and the target temperature of the MHT group was 32-36 ℃. In the sham group, only conventional tube placement was performed, and the target temperature was 37 ℃. At 24 h after resuscitation, neurologic deficit score (NSD) was evaluated. Animals were then euthanized and venous blood sample was taken to detect brain injury and immune function indicators. Results Compared with the sham group, the NDS score increased in the NT group and the MHT group, and expression levels of neuron specific enolase (NSE) and S100B protein increased (P<0.05). Compared with the NT group, the NDS score, NSE and S100B protein expression levels were decreased in the MHT group (P<0.05). There were no significant differences in levels of IgM, IgA, IgG and T cell subsets CD4, CD8, CD4/CD8 between the groups. Conclusion Mild hypothermia treatment can significantly improve NDS, reduce brain injury after CA and has no significant effect on immune function.

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Effects of Mycoplasma pneumoniae community acquired respiratory distress syndrome toxin on monocyte subsets

ZHANG Cen, WANG Zhihua, YANG Lei

2023, 51 (10): 1080-1083. doi: 10.11958/20230118

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Objective To investigate the effect of Mycoplasma pneumoniae community-acquired respiratory distress syndrome (CARDS) toxin on peripheral blood mononuclear cell subsets in mice. Methods The recombinant CARDS toxin was prepared. Thirty-two mice were divided into the model group and the normal control group according to random number table with 16 mice in each group. Each mouse in the model group was injected with 40 μL PBS containing 50 μg of CARDS toxin. In the normal control group, the same dose of PBS was injected into trachea. Blood samples were collected from abdominal aorta 24 h after the modeling, and the proportion of peripheral monocyte subsets (CD11b⁺Ly6G^-Ly6C⁺) was detected by flow cytometry. The mice were sacrificed, and lung histopathological changes were observed by HE staining, and the lung tissue injury score was performed. The expression levels of interleukin (IL)-1β and IL-18 in lung tissue were detected by Real-time PCR. The levels of IL-1β and IL-18 in peripheral blood were determined by enzyme-linked immunosorbent assay. Results Compared with the normal control group, CD11b⁺Ly6G^-Ly6C⁺ratio of mouse monocytes was increased in the model group (P<0.05). HE staining showed that alveolar wall thickened, inflammatory cell infiltration increased, pathological score increased significantly (P<0.01), the expression levels of IL-1β and IL-18 mRNA in lung tissue and contents of inflammatory cytokines IL-1β and IL-18 in serum increased significantly (P<0.01). Conclusion The increased proportion of Ly6C⁺ monocytes in mice induced by CARDS toxin may be closely related to the incidence of Mycoplasma pneumoniae pneumonia.

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Impact of loganin on pain response in rats with lumbar disc herniation by inhibiting CXCL12/CXCR4 signal pathway

QIAO Song, HU Yanping, HE Shenghua

2023, 51 (10): 1084-1089. doi: 10.11958/20230249

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Objective To investigate the impact of loganin on pain response in rats with lumbar disc herniation (LDH) by regulating CXCL12/CXCR4 signal pathway. Methods Ten of the 50 rats were randomly selected as the sham group according to random number table method, and LDH rat model was established in other rats. Model rats were randomly divided into the model group, the CXCR4 antagonist group (AMD3100 group, intrathecal injection, 20 μg), the loganin group (5 mg/kg, intraperitoneal injection), and the loganin (5 mg/kg, intraperitoneal injection)+CXCL12 group (intrathecal injection, 250 ng), with 10 animals in each group. After the intervention according to drug groups, the latency of thermal foot withdrawal reaction and the threshold of mechanical foot withdrawal threshold were measured respectively. HE staining was applied to observe histomorphological changes of spinal dorsal horn. Levels of TNF-α, IL-1β and IL-6 in spinal dorsal horn were detected by ELISA. Immunofluorescence staining was applied to detect the activation of microglia. Western blot assay was used to detect the level of CXCL12/CXCR4 pathway related proteins. Results Compared with the sham group, the model group had severe spinal cord dorsal horn tissue injury, the latency of thermal foot withdrawal reaction and mechanical foot withdrawal threshold were lower, and levels of microglia, levels of TNF-α, IL-1β, IL-6, and protein expression levels of CXCL12, CXCR4, NLRP3, caspase-1 and IL-1β in spinal dorsal horn tissue were higher (P<0.05). Compared with the model group, the pathological injury of spinal dorsal horn tissue was reduced in the AMD3100 group and the loganin group. The latency of thermal foot withdrawal reaction and mechanical foot withdrawal threshold were higher, and levels of microglia, levels of TNF-α, IL-1β, IL-6, and the protein expression levels of CXCL12, CXCR4, NLRP3, caspase-1 and IL-1β in the spinal dorsal horn tissue were also lower (P<0.05). CXCL12 could weaken the improvement effect of loganin on pain response in LDH rats (P<0.05). Conclusion Loganin can improve the pain response of LDH rats, which may be related to the inhibition of CXCL12/CXCR4 signal pathway.

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Effect and its mechanism of emodin on the ferroptosis of chondrocytes in rats with knee osteoarthritis

WANG Ke, YE Hanlu

2023, 51 (10): 1090-1097. doi: 10.11958/20221976

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Objective To investigate the effect and its mechanism of emodin on the ferroptosis of chondrocytes in rats with knee osteoarthritis (KOA). Methods A total of 96 SD rats were divided into the sham operation (sham) group, the KOA group, the emodin (EMO) group and the emodin+Nrf2 inhibitor (EMO+ML385) group, with 24 rats in each group. Except for the sham group, KOA rat models were established by modified Hulth method in each group. The tumor necrosis factor-α (TNF-α), nitric oxide (NO) and prostaglandin E2 (PGE2) levels were detected by enzyme-linked immunosorbent assay (ELISA). General morphology of knee joint was observed with naked eyes. The pathological morphology of knee joint cartilage was observed with safranine O-solid green staining. Apoptosis rate of chondrocytes of knee joint was detected by TdT-mediated dUTP nick end labeling (TUNEL) staining. Malondialdehyde (MDA), reactive oxygen species (ROS), glutathione (GSH) and ferrous ion (Fe²⁺) levels in knee joint cartilage were detected by biochemical reagent kit. Expression levels of matrix metalloproteinase (MMP)-3, MMP-13, collagen type II α1 (COL2A1), prostaglandin-endoperoxide synthase 2 (PTGS2), glutathione peroxidase 4 (GPX4), acyl-CoA synthetase long chain family member 4 (ACSL4), nuclear factor E2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) mRNA or protein in knee joint cartilage were detected by real-time fluorescence quantitative PCR (qRT-PCR), Western blot assay or immunohistochemistry. Results Compared with the sham group, the serum TNF-α, NO and PGE2 levels, Osteoarthritis Research Society International (OARSI) score, apoptosis rate, mRNA expression of MMP-3, MMP-13 and PTGS2, MDA, ROS, and Fe²⁺ levels, proportion of ACSL4 positive cells, and protein expression of ACSL4, nuclear Nrf2 and HO-1 increased in the KOA group. GSH levels, mRNA expression of COL2A1, proportion of GPX4 positive cells, and protein expression of GPX4 and cytoplasmic Nrf2 decreased (P<0.05). The articular cartilage was obviously damaged. EMO can improve the inflammatory reaction, knee joint cartilage tissue injury and chondrocyte ferroptosis of KOA rats, and further activate Nrf2/HO-1 signal pathway. ML385 not only inhibited the activation of Nrf2/HO-1 signal pathway, but also attenuated the improvement of EMO on knee joint cartilage tissue injury and chondrocyte ferroptosis of KOA rats. Conclusion Emodin can activate Nrf2/HO-1 signal pathway, inhibit chondrocyte ferroptosis and protect knee articular cartilage of KOA rats.

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The improvement effect of astaxanthin on rats with polycystic ovary syndrome by regulating Klotho expression and Wnt/β-Catenin signaling pathway

SONG Yu, ZHU Zhengyan, HUANG Huan, LI Chun, LU Jingquan, LIU Heyu, HU Lina

2023, 51 (10): 1098-1103. doi: 10.11958/20221727

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Objective To study the improvement mechanism of astaxanthin in rats with polycystic ovary syndrome (PCOS) by regulating Klotho expression and Wnt/β-Catenin signaling pathway. Methods Thirty PCOS model rats were constructed by intragastric administration of letrozole, and rats were randomly grouped into 6 groups: the model group, the low-dose astaxanthin group (50 mg/kg), the middle-dose astaxanthin groupm (100 mg/kg), the high-dose astaxanthin group (200 mg/kg), the high-dose astaxanthin+no-load group and the high-dose astaxanthin+Klotho knockdown group, with 5 rats in each group. Another 5 SD rats were taken as the control group. After treatment, the body weight and ovarian weight of rats in each group were measured. Forty-five SD rats were modeled in the same way and randomly grouped into: the model group, the astaxanthin group (200 mg/kg), the astaxanthin+Klotho knockdown group, the astaxanthin+lithium chloride group and the astaxanthin+PD98059 (MEK inhibitor) group, with 9 rats in each group. Another 9 SD rats were taken as the control group. After treatment, data of body weight, ovarian weight and volume, the number of cystic follicles in ovarian tissue, levels of serum hormones including follicle-stimulating hormone (FSH), luteinizing hormone (LH), testosterone (T) and levels of inflammatory factors prostaglandin 2 (PGE2), interleukin (IL)-17, tumor necrosis factor-α (TNF-α), and IL-18, the expression of Klotho protein and the expression of Wnt/β-Catenin and MEK/ERK signal-related proteins in ovarian tissue were detected in rats of each group. Results Compared with the control group, body weight and ovarian weight were obviously increased in the model group (P<0.05). Compared with the model group, body weight and ovarian weight were all decreased in the low-dose astaxanthin group, the middle-dose astaxanthin group and the high-dose astaxanthin group in a dose-dependent manner (P<0.05). Compared with the high-dose astaxanthin group, body mass and ovarian mass of rats were increased in the high-dose astaxanthin+Klotho knockdown group (P<0.05). There was no significant change in each index of rats in the high dose of astaxanthin+empty load group (P>0.05). Compared with the control group, serum level of FSH, and expression levels of Klotho, p-MEK, p-ERK1/2 proteins in ovarian tissue were obviously decreased in the model group (P<0.05). Body weight, ovarian mass and volume, number of cystic follicles, serum levels of LH, T, PGE2, IL-17, TNF-α and IL-18, and expression levels of Wnt1 and β-Catenin proteins in ovarian tissue were obviously increased in the model group (P<0.05). Compared with the model group, the level of FSH, the expression levels of Klotho, p-MEK and p-ERK1/2 proteins in ovarian tissue were increased in the astaxanthin group (P<0.05). Body weight, ovarian mass and volume, number of cystic follicles, serum levels of LH, T, PGE2, IL-17, TNF-α and IL-18, and expression of Wnt1 and β-Catenin proteins in ovarian tissue were decreased in the astaxanthin group (P<0.05). Knockdown of Klotho, lithium chloride and PD98059 can weaken the improvement effect of astaxanthin on PCOS rats. Conclusion Astaxanthin can down-regulate Wnt/β-Catenin signaling and activate MEK/ERK signaling by up-regulating Klotho, thereby preventing inflammation, improving hormone levels in PCOS rats, reducing number of cystic follicles, and ultimately alleviating the symptoms of polycystic ovaries.

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Effects of berberine on Th17/Treg balance in autoimmune prostatitis rats by regulating AMPK/NF-κB signaling pathway

MIAO Ruiyu, WANG Bi, LONG Zongmin

2023, 51 (10): 1104-1109. doi: 10.11958/20221816

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Objective To investigate the influence of berberine on T helper cell 17 (Th17)/regulatory T cell (Treg) balance in autoimmune prostatitis (EAP) rats by regulating adenylate-activated protein kinase (AMPK)/nuclear factor-κB (NF-κB) signaling pathway. Methods Ninety SD male rats were randomly divided into the control group (normal feeding), the model group, the low-dose berberine group (25 mg/kg), the high-dose berberine group (50 mg/kg), the positive control group (40 mg/kg celecoxib) and the inhibitor group (50 mg/kg berberine + 0.2 mg/kg AMPK inhibitor compound C), with 15 rats in each group. Except for the control group, rats in other groups were all injected with complete Freund's adjuvant and prostate tissue mixed suspension in pelvic region and bilateral scapula to construct the EAP rat model. After successful modeling, drug treatment was carried out by gavage, once a day for four weeks. HE staining was used to observe the pathological conditions of prostate tissue in each group. Immunohistochemical method was used to detect the expressions of retinoic acid-related nuclear orphan receptor γt (RORγt) and forkhead box protein 3 (Foxp3) in rat prostate tissue. ELISA method was used to detect serum levels of interleukin (IL)-17 and IL-10 in rats. Flow cytometry was used to detect the ratio of Th17/Treg cells in peripheral blood of rats. Western blot assay was used to detect the expression of AMPK/NF-κB signaling pathway-related proteins in rat prostate tissue. Results Compared with the model group, there were pink secretions in prostate tissue cavity of rats in the low-dose and the high-dose berberine groups and the positive control group. The glandular epithelial cells were neatly arranged, and the inflammation subsided obviously. Serum IL-17 levels, peripheral blood Th17/Treg ratio, and RORγt and p-NF-κB p65/NF-κB p65 expression levels in prostate tissue were decreased, and serum IL-10 level, Foxp3, p-AMPK/AMPK and IκBα expression levels in prostate tissue were increased (P<0.05). After adding AMPK inhibitor compound C, the promoting effect of berberine on Th17/Treg balance in EAP rats was attenuated. Conclusion Berberine can promote Th17/Treg balance in EAP rats by regulating AMPK/NF-κB signaling pathway.

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The influence of blood pressure fluctuation on poor prognosis in patients with severe aortic stenosis

YANG Xueyuan, FENG Chao, FENG Jinping

2023, 51 (10): 1110-1116. doi: 10.11958/20222067

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Objective To analyze risk factors for the poor prognosis of severe aortic stenosis (AS), and to evaluate the influence of blood pressure and blood pressure fluctuation on the poor prognosis of severe AS. Methods Clinical data of patients with acute decompensation of chronic heart failure (CHF) admitted to the Cardiac Intensive Care Unit of Tianjin Chest Hospital from December 2018 to March 2022 were collected. Patients were divided into the AS group and the non-NAS group according to aortic stenosis lesions. Baseline clinical data, follow-up results, blood pressure for consecutive 3 days after admission and blood pressure monitoring values for 2 days before discharge were collected, and indexes such as mean arterial pressure (MAP), systolic pressure difference (?SBP) and diastolic pressure difference (?DBP) were calculated. The primary endpoint event was a composite endpoint of rehospitalization and/or all-cause death due to heart failure, and the secondary endpoint event was cardiogenic death. The Kaplan-Meier survival curve was used to compare the survival differences between the two groups. Univariate and multivariate Cox regression models were used to analyze prognostic factors of aortic stenosis in patients with AS. Further subgroup analysis was conducted according to whether the AS group was complicated with hypertension and the ?SBP quartile to compare the occurrence of end events and outcome differences between groups. Results Ninety-one patients were included, including 50 in the AS group and 41 in the NAS group. Compared with the NAS group, the AS group was older, and had a higher proportion of smoking history, diabetes history, and lower extremity edema symptoms on admission, lower albumin level, and higher pulmonary artery systolic blood pressure (P<0.05). Kaplan-Meier results showed that the incidence of primary (P=0.030) and secondary (P=0.037) endpoint events were higher in the AS group than that in the NAS group. Cox regression results showed that old age, combined hypertension, elevated ?SBP, and elevated NE were independent risk factors for primary endpoint events (P<0.05), and old age and elevation of ?SBP and ?DBP were independent risk factors for secondary endpoint events (P<0.05). Subgroup analysis showed that the incidence of primary endpoint events was higher in AS patients with hypertension than that in AS patients with non-hypertension patients (P=0.011). The larger the ?SBP, the higher the incidence of primary endpoint events in the AS group (P<0.001). Conclusion Hypertension, elevated ?SBP and old age are independent risk factors for poor prognosis of AS. Patients with AS have lower survival rates. The greater the fluctuation of blood pressure, the lower the survival rate and the worse the prognosis of AS patients.

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Predictive value of prognosis of prealbumin dynamics in the treatment of malignant obstructive jaundice after biliary stent implantation

HUANG Yi, GUAN Min, ZHANG Xue, ZHANG Yong, TANG Shanhong

2023, 51 (10): 1117-1121. doi: 10.11958/20230078

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Objective To investigate the predictive value of dynamic changes in serum prealbumin (PAB) in the treatment of malignant obstructive jaundice (MOJ) with the first endoscopic retrograde cholangiopancreatography (ERCP) biliary stenting. Methods The clinical data of 119 patients with MOJ who were relieved by ERCP stenting for the first time were retrospectively analyzed. Patients were divided into the survival group and the death group according to whether their survival period was greater than 1 year. The liver function indexes of patients before and 7 days after surgery were collected, and the dynamic change rate of PAB (7 d after surgery/preoperative) was calculated. The receiver operating characteristic (ROC) curve was used to determine the best cut-off value of the dynamic change rate of PAB. Patients were divided into two groups according to the cut-off value. The survival curves of the two groups were plotted by K-M method, and risk factors affecting the long-term survival prognosis of patients were analyzed by the Cox proportional hazard model. Results The 1-year survival rate of 119 patients was 45.4% (54/119) and the case fatality rate was 54.6% (65/119). The dynamic rate of PAB was 1.1 (0.9, 1.4) in the survival group, and 0.9 (0.7, 1.0) in the death group, which was significantly higher in the survival group than that in the death group (P<0.05). ROC analysis showed that the area under the ROC curve was 0.799（95% CI：0.704-0.895）and the optimal cut-off value of PAB dynamic rate was 1.008, and the survival analysis showed that the one-year survival rate of the group with high PAB dynamic rate was significantly higher than that in the group with low PAB dynamic rate (P<0.01). Multivariate analysis showed that low dynamic rate of PAB was an independent risk factor for long-term survival of patients with MOJ after ERCP. Conclusion The dynamic rate of PAB can be used as a potential prognostic indicator for MOJ patients.

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Influencing factors and construction of risk prediction model of immune thrombocytopenia in children

ZHOU Nina, GU Jianhui, YANG Zhiping, SHENG Yu, JIANG Rong

2023, 51 (10): 1122-1125. doi: 10.11958/20221631

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Objective To study factors influencing the outcome of immune thrombocytopenia (ITP) in children, and construct a risk prediction model. Methods A total of 166 children with ITP were included in this study. Clinical characteristics and laboratory indicators of children with different outcomes were comparatively analyzed. Multivariate Logistic regression analysis was performed to screen factors influencing the outcome of children with ITP, and a risk prediction model was constructed based on analysis results. Hosmer-Lemeshow tested the goodness of fit of the model. The receiver operating characteristic (ROC) curve was plotted to analyze the predictive performance of the model. Results There were 118 children with good outcomes and 48 children with poor outcomes. Compared with the good outcome group, the proportions of age ≥1, ANA positive at first diagnosis and blood urea nitrogen (BUN) level were higher in the poor outcome group, while the proportion of platelet count (PLT) ≥20×10⁹/L at first diagnosis, absolute lymphocyte count (ALC) at first diagnosis and the percentage of megakaryocytes in bone marrow at first diagnosis were lower in the poor outcome group (P<0.05). Multivariate Logistic regression analysis showed that age ≥1 (95% CI: 1.358-83.563) and higher BUN level (95% CI: 4.013-32.615) were risk factors for disease outcome in children with ITP. PLT ≥20×10⁹/L at initial diagnosis (95% CI: 0.036-0.519) and higher ALC at initial diagnosis (95% CI: 0.197-0.571) were protective factors for disease outcome in children with ITP. Homser-lemeshow test showed χ²=8.486, P=0.387. The AUC of this model in predicting the outcome of children with ITP was 0.963 (95% CI: 0.934-0.991, P<0.001), and the sensitivity and specificity were 95.83% and 88.98%. Conclusion Age ≥1 and higher BUN level are risk factors for disease outcome in children with ITP. PLT ≥20×10⁹/L at initial diagnosis and higher ALC level at initial diagnosis are protective factors. The risk prediction model constructed according to these influencing factors has good predictive valuse for disease outcome of children with ITP.

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Comparison of stress response and postoperative analgesia of ultrasound-guided paravertebral block and erect spinae plane block in patients undergoing video-assisted thoracic surgery

HU Yang, ZHANG Lili, ZHANG Ye

2023, 51 (10): 1126-1130. doi: 10.11958/20222027

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Objective To evaluate the effects of ultrasus-guided thoracic paravertebral nerve block (TPVB) and erectile spinal-plane block (ESPB) on perioperative stress response and analgesia in patients undergoing thoracoscopic surgery. Methods Ninety patients underwent thoracoscopic surgery under elective general anesthesia were included and randomly divided into the control group (group C), the ESPB group (group E) and the TPVB group (group T), with 30 cases in each group. Before anesthesia induction, no treatment was given to the group C, and the group E and the group T were treated with ultrasound guided ESPB and TPVB on the affected side, and injected with 20 mL of 0.5% ropivacaine. Patient controlled analgesia (PCIA) was connected after operation. The venous blood samples were collected immediately before operation (T₀), 30 minutes after operation (T₁) and at the time of tracheal catheter removal (T₂) to test the concentration of blood glucose (Glu) and serum cortisol (Cor). The type of surgery, the duration time of surgery, the number of patients who needed to use phenylephrine and the amount of remifentanil for hypotension during operation were recorded. Visual analog scores (VAS) of patients at rest and coughing status at T₂, 12 hours after operation (T₃), 24 hours after operation (T₄), 48 hours after operation (T₅), the total number of PCIA compression and the total amount of PCIA sufentanil infusion within 48 hours after operation, the use of rescue analgesia, and the occurrence of adverse reactions were recorded. Results Compared with T₀, concentrations of Cor and Glu at T₁ and T₂, were increased in patients (P<0.05). Compared with the group C, Glu was significantly decreased in the group T, the resting VAS pain scores of T₂, T₃, T₄ and T₅ were decreased in the group T and the group E. The coughing VAS pain scores of T₂ and T₄ were decreased, and the intraoperative dosage of reifentanil, the number of PCIA compression and the total infusion of sufentanil were decreased. The remedial analgesic rate was decreased in the T group (P<0.05). Compared with the group T, the resting VAS scores of T₂, T₃ and T₄ were increased in the group E, and the coughing VAS scores of T₃ were increased, the number of PCIA compression, remifentanil usage and the total infusion of PCIA sufentanil were increased. The percentage of intraoperative use of norepinephrine for hypotension was decreased (P<0.05). Conclusion TPVB can inhibit the stress response of patients undergoing thoracoscopic surgery. TPVB could provide better postoperative analgesia than ESPB, and has a significant inhibitory effect on intraoperative blood pressure.

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Influence of body mass index on postoperative complications and long-term prognosis in patients with esophageal squamous cell carcinoma

ZHAO Xuenan, GUO Xufeng, DING Chengzhi, CHEN Dongdong, WEI Lirui, LI Chen, QIN Guijun, WANG Jiao

2023, 51 (10): 1131-1135. doi: 10.11958/20230090

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Objective To investigate the influence of body mass index (BMI) on postoperative complications and long-term prognosis in patients with esophageal squamous cell carcinoma (ESCC). Methods A total of 1 479 patients with ESCC who underwent radical resection were included in this study. According to BMI, 100 patients were divided into the low BMI group (BMI<18.5 kg/m²), 850 cases were divided into the normal BMI group (18.5 kg/m²≤BMI<24 kg/m²) and 529 cases were divided into the high BMI group (BMI≥24 kg/m²). Postoperative complications of patients were compared between the three BMI groups. Kaplan-Meier survival curve was used to compare the overall survival of the three groups, and Cox proportional risk model was used to analyze the influence of BMI on prognosis of patients. Results Compared with the normal BMI group, the incidence of incision infection was increased in the high BMI group, and the incidence rates of pulmonary infection and arrhythmia were increased in the low BMI group (P<0.05). Compared with the low BMI group, the incidence of arrhythmia was lower in the high BMI group (P<0.05). There were no significant differences in the incidence of anastomotic leakage, respiratory failure, chylothorax and vocal cord paralysis between the three groups (P>0.05). Kaplan-Meier survival curve showed that the 5-year cumulative overall survival rate of patients was significantly lower in the low BMI group than that in the normal BMI group (Log-rank χ²=16.857, P<0.01) and the high BMI group (Log-rank χ²=18.802, P<0.01), while there was no significant difference in 5-year cumulative overall survival rate between the high BMI group and the normal BMI group (Log-rank χ²=0.324, P=0.569). Multivariate Cox regression analysis showed that low BMI was an independent risk factor for the prognosis of patients with esophageal squamous cell carcinoma (HR=1.449, 95% CI: 1.084-1.938, P<0.05). Conclusion High BMI increases the incidence of incision infection after esophagectomy in patients with ESCC. Low BMI increases the incidence of pulmonary infections and arrhythmia, and is an independent predictor for poor overall survival.

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Evaluation value of QRS complex time limit combined with serum angptl2, angptl7 and sST2 for short-term prognosis of patients with acute heart failure

JIANG Meiyan, HUANG Henggui, YUAN Shiguo

2023, 51 (10): 1136-1140. doi: 10.11958/20230053

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Objective To investigate the prognostic value of QRS complex duration combined with serum angiopoietin-like protein (angptl)2, angptl7 and soluble tumor suppressor factor 2 (sST2) in patients with acute heart failure (AHF). Methods A total of 104 patients with AHF were collected as the case group, and their baseline data were recorded. According to the American New York Heart Association (NYHA) classification, patients were divided into the class Ⅱ group, the class Ⅲ group and the class Ⅳ group. According to the prognosis, patients were grouped into the good prognosis group and the poor prognosis group, and 98 healthy subjects in our hospital were collected as the control group. The QRS complex time limit was measured by electrocardiograph, and serum levels of angptl2, angptl7 and sST2 were detected by enzyme-linked immunosorbent assay. Receiver operating characteristic (ROC) curve was applied to analyze the predictive value of QRS complex duration combined with serum angptl2, angptl7 and sST2 levels on the prognosis of patients with AHF. Multivariate Cox regression was used to analyze influencing factors of poor prognosis in patients with AHF. Results The QRS complex time limit and serum levels of angptl2, angptl7 and sST2 were higher in the case group than those in the control group (P<0.05). The QRS complex time limit and serum levels of angptl2, angptl7 and sST2 in the grade Ⅱ group, the grade Ⅲ group and the grade Ⅳ group were increased in turn (P<0.05). The NYHA classification, left ventricular end diastolic dimension (LVEDD), QRS complex time limit and serum levels of angptl2, angptl7 and sST2 were greatly higher in the poor prognosis group than those in the good prognosis group, and the left ventricular ejection fraction (LVEF) was greatly lower in the poor prognosis group than that in the good prognosis group (P<0.05). The AUC of QRS complex time limit, serum angptl2, angptl7, sST2 and the combined prediction of poor prognosis in patients with AHF were 0.773, 0.767, 0.833, 0.743 and 0.907, respectivily. High NYHA class and high sST2 were independent risk factors for poor prognosis in patients with AHF (P<0.05). Conclusion The QRS complex duration, serum levels of angptl2, angptl7 and sST2 in AHF patients increase with the worsening of the disease. NYHA grading and sST2 are independent factors for poor short-term prognosis in AHF patients.

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Curative effect of butylphthalide combined with low-dose recombinant tissue-type plasminogen activator for intravenous thrombolytic therapy in elderly patients with ultra-early cerebral infarction

ZHOU Zeng, FENG Zhaohai, XU Jing, ZHENG Yingwei, DING Han, LI Meiying

2023, 51 (10): 1141-1145. doi: 10.11958/20230414

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Objective To explore the curative effect of butylphthalide (D) combined with low-dose recombinant tissue-type plasminogen activator (rt-PA) for intravenous thrombolysis on elderly patients with ultra-early cerebral infarction. Methods A total of 156 elderly patients with ultra-early cerebral infarction were retrospectively enrolled as the research objects. According to different treatment methods, patients were divided into the D+low rtPA group [30 cases, butylphthalide combined with low-dose (0.6 mg/kg) rt-PA], the D+standard rtPA group [62 cases, butylphthalide combined with standard-dose (0.9 mg/kg) rt-PA], the low rtPA group [30 cases, low-dose (0.6 mg/kg) rt-PA] and the standard rtPA group [34 cases, standard-dose (0.9 mg/kg) rt-PA]. After thrombolysis, scores of National Institutes of Health Stroke Scale (NHISS), Modified Rankin Scale (mRS) and activity of daily living (ADL), and inflammation indexes were compared between the four groups. The poor prognosis was statistically analyzed. Results After 7 days of thrombolysis, the addition of butylphthalide reduced NHISS and mRS scores, as well as neutrophil count (NET), platelet count (PLT) and neutrophil absolute value/lymphocyte absolute value (NLR) levels (P<0.05), while increased ADL scores and lymphocyte count (LYT, P<0.05). The levels of NHISS score, mRS score, ADL score, NET, LYT, PLT, NLR and platelet absolute value/neutrophil absolute value (PNR) were not affected by different doses of rt-PA (P>0.05). There was no significant difference in the overall adverse prognosis rate between the four groups (P>0.05). Conclusion Butylphthalide combined with low-dose and standard-dose rt-PA is safe and effective in the treatment of elderly patients with ultra-early cerebral infarction.

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Effect of epidural injection with different doses of 1.5% chloroprocaine in relieving breakthrough pain during labor analgesia

JI Tianzhen, XU Cheng, LIU Hongxia, LI Guofang, LI Rui, ZHU Haijuan

2023, 51 (10): 1146-1152. doi: 10.11958/20230652

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Objective To investigate the effect of epidural injection of 1.5% chloroprocaine on breakthrough pain during labor analgesia. Methods A total of 128 primiparas with breakthrough pain after epidural labor analgesia were randomly divided into four groups (R, C₁, C₂ and C₃groups). When breakthrough pain occurred, 0.15% ropivacaine 6 mL was injected epidural in the group R, and 1.5% chloroprocaine 6, 8 and 10 mL were injected epidural in the group C₁, the group C₂ and the group C₃, respectively. The visual analogue score (VAS) for pain was assessed every 3 minutes after the administration of local anesthetics. The primary outcome was recorded including the onset time of breakthrough pain after the administration and the success rate of breakthrough pain treatment. Secondary outcome included rescue analgesia rate, maternal satisfaction (5 minutes after administration of breakthrough pain, 24 hours after the delivery), VAS, modified Bromage score, fetal heart rate (FHR) at first breakthrough pain (T₀), 3 min (T₁), 6 min (T₂), 9min (T₃), 12 min (T₄), 15 min (T₅) and 18 min (T₆) after administration, the interval and duration of uterine contraction at the onset time of breakthrough pain, the second, fourth, sixth, eighth, tenth and twelfth contractions after administration, Apgar score in 1 and 5 min and adverse reactions. Results Kaplan-Meier survival analysis showed that the median onset time (95%CI) of breakthrough pain in the group R, the group C₁, the group C₂ and the group C₃ were 14.3 (13.4-15.2) min, 6.6 (5.6-7.6) min, 5.9 (5.0-6.8) min and 4.7 (3.9-5.4) min, respectively. The onset time of administration was significantly faster in the group C₃ than that in the group C₁ and the group C₂ (Log-rank χ² was 9.852 and 5.325, P<0.05). Compared with the group R, the success rate of breakthrough pain treatment and maternal satisfaction (5 minutes after administration, 24 hours after the delivery) were significantly increased in the group C₂ and the group C₃, and remedial analgesia rate was significantly decreased in the group C₃ (P < 0.01). Compared with the group C₁, the success rate of breakthrough pain treatment was increased and the remedial analgesia rate was decreased in the group C₃ Maternal satisfaction (5 minutes after administration, 24 hours after the delivery ) was significantly increased in the group C₂ and the group C₃(P < 0.01). At T₁, the VAS score was significantly lower in the group C₂ and the group C₃ than that in the group R, and which was significantly lower in the group C₃than that in the group C₁ (P < 0.05). At T_2~5, the VAS score was significantly lower in the group C₁, the group C₂ and the group C₃ than that in the group R, and which was significantly lower in the group C₂ and the group C₃ than that in the group C₁ (P < 0.05). At T₆, VAS scores were significantly lower in the group C₂ and the group C₃ than those of the group R and the group C₁ (P<0.05). There were no significant differences in the interval of uterine contractions and duration of uterine contractions between the four groups. Compared with the group R and the group C₁, the probability of lower limb numbness within 30 minutes after administration of breakthrough pain was significantly higher in the group C₃(P<0.05). There were no significant differences in modified Bromage score, FHR at different time points before and after administration, and Apgar score at 1 and 5 min between the four groups. Conclusion The optimal effective dose of epidural injection of 1.5% chloroprocaine for the treatment of breakthrough pain during labor analgesia is 8 mL.

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