Tianjin Medical Journal

Study on the effect of Chlorella extract on promoting skin wound healing in diabetic mice

HUANG Yu, HE Ruiying, LIU Sen, CHEN Kaiyuan, LI Meiyun, CHENG Jianye, WU Yan

2024, 52 (4): 337-345. doi: 10.11958/20230825

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Objective To investigate the effect of Chlorella vulgaris extract (CE) on skin wound healing in diabetic mice. Methods The blank control group (CON group), CE low (1 mg/L), medium (10 mg/L) and high (100 mg/L) dose groups were set up in vitro cytotoxicity assay. The additional H₂O₂(100 μmol/L) group was set up for antioxidant experiments, and the concentration of H₂O₂ (100 μmol/L) in each experimental group was also added. The additional lipopolysaccharide (1 mg/L) group was set up for the anti-inflammatory experiments, and the concentration of lipopolysaccharide (1 mg/L) in each experimental group was also added. The cytotoxicity of CE was detected by MTT method. The antioxidant activity of CE was investigated by fluorescent probe assay. The anti-inflammatory effect of CE was detected by fluorescent quantitative PCR. The blank control group (CON group), CE low (0.5 g/L), medium (5 g/L) and high (50 g/L) dose groups were set up, and the antibacterial properties of CE were examined by plate colony counting method. The diabetic mouse model was induced by streptozotocin, and model mice were divided into the control group, the CE low (0.5 g/L), medium (5 g/L) and high (50 g/L) dose groups using the random number table method, with 12 mice in each group. After the model was successfully established, two 6 mm wounds were symmetrically created on each side of spine of mouse. The wounds were administered once daily for a total of 14 consecutive days. The wound healing was observed and photographed on days 0, 7 and 14 respectively. The wound samples were taken and processed for histology (7 d and 14 d) and immunofluorescence (7 d). Results In vitro experiments: CE low, medium and high dose groups showed good cytocompatibility with NIH-3T3 cells compared to the control group, and CE low, medium and high dose groups showed weaker reactive oxygen species (ROS) fluorescence signal compared to the H₂O₂ group. CE low, medium and high dose groups showed lower expression of CD86 and higher expression of CD206 compared to the control group (P<0.05). The inhibition of bacterial growth was enhanced in the CE low, medium and high dose groups compared to the control group (P<0.05). In vivo experiment results demonstrated that wound healing rates on the 7th and 14th day after operation were accelerated in the CE group, and the wound healing effect was obvious. Results of HE and Masson staining showed that there were more collagen deposition in wound in the CE low, medium and high dose groups compared to the control group. Results of the immunofluorescence assay showed that the higher expression levels of α-smooth muscle actin (α-SMA), platelet endothelial cell adhesion molecule-1 (CD31) and macrophage mannose receptor (CD206) after treatment, and the lower levels of leukocyte differentiation antigen 86 (CD86) in wound compared to the control group. Conclusion CE can effectively promote wound healing in diabetic mice, and the mechanism may be related to its antioxidant, anti-inflammatory and antibacterial effects.

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The effect of ADAM8 expression on acute alcoholic liver injury in mice

YANG Mengli, LI Sanqiang, ZHANG Kaijie, CUI Qinyi, FENG Jiayang, LI Yilin, LI Haoyuan, QI Jinghan

2024, 52 (4): 346-349. doi: 10.11958/20231741

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Objective To investigate the role and molecular mechanism of CRISPR/Cas9 technology targeting the inhibition of disintegrin and metalloprotease 8 (ADAM8) in acute alcoholic liver injury in mice. Methods Eighteen healthy Kunming female mice aged 6-8 weeks were randomly divided into the normal group, the alcohol group and the plasmid group, with 6 mice in each group. The normal group was given no treatment, and the plasmid group was injected with three-in-one recombinant plasmid ADAM8-sgRNA3 (3 g/kg) by CRISPR/Cas9 technology to inhibit the ADAM8 gene using hydrodynamic injection tail vein method. The alcohol group was injected with an equal amount of physiological saline through tail vein. After 3 days of regular feeding, the alcohol group and the plasmid group were subjected with 50% (V/V) alcohol analytical grade ethanol by a one-time gavage (14 mL/kg) to induce acute liver injury. After fasting for 16 hours, eyeball blood sample was taken to detect activities of alanine aminotransferase (ALT) and aspartate aminotransferase (AST). Mice were euthanized and liver tissue was separated and extracted. Hematoxylin-Eosin staining (HE staining) and Periodic Acid-Schiff staining (PAS staining) were used to detect liver injury and glycogen changes in each group of mice. The expression levels of ADAM8, cytochrome CYP450 2E1 (CYP2E1), heat shock protein 70 (HSP70) and mitogen-activated protein kinase (MAPK) pathway related factors were detected by Western blot assay. Results Compared with the normal group, ALT and AST were increased, liver injury score was increased, glycogen content was decreased, ADAM8, CYP2E1, phosphorylated extracellular regulated kinase 1/2 (p-ERK1/2), phosphorylated p38 (p-p38) MAPK and phosphorylated c-Jun aminoterminal kinase (p-c-Jun) were increased in the alcohol group. The expression of HSP70 was decreased (P<0.05). Compared with alcohol group, ALT and AST were decreased, liver injury score was decreased in the plasmid group (P<0.05). Glycogen content was increased. ADAM8, CYP2E1, p-ERK1/2, p-P38 MAPK and p-c-Jun expression levels were decreased, while HSP70 expression was increased (P<0.05). Conclusion Targeted inhibition of ADAM8 expression by CRISPR/Cas9 technology can improve acute alcoholic liver injury in mice through MAPK signaling pathway.

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Influence of Yiqi Shengqing recipe on neuron pyroptosis in ischemic stroke rats by regulating HIF-1α/NLRP3 signal pathway

WANG Yue, QUAN Xingmiao, WANG Yu, SONG Chunxia, SHAO Yue, XU Liwei

2024, 52 (4): 350-355. doi: 10.11958/20231141

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Objective To investigate the influence of Yiqi Shengqing recipe on neuronal pyroptosis in ischemic stroke rats by regulating hypoxia inducible factor-1α (HIF-1α)/NOD-like receptor thermal protein domain associated protein 3 (NLRP3) signal pathway. Methods SD rats were randomly divided into the sham operation group (S group), the model group (M group), the low, medium and high dose (3.465, 6.930 and 13.860 g/kg) Yiqi Shengqing recipe groups, and high-dose Yiqi Shengqing recipe+HIF-1α activator DMOG (13.860 g/kg Yiqi Shengqing recipe+40 mg/kg DMOG) group, 15 animals in each group. The stroke model was constructed by thread plug method. After successful modeling, neural function defects were evaluated including TTC staining for evaluating the volume of cerebral infarction, ELISA for detecting the serum interleukin (IL)-1β and IL-18 levels, HE staining for detecting pathological changes of ischemic cortex, TUNEL staining for detecting neuronal apoptosis and Western blot assay for detecting the expression of pyroprotein and HIF-1α/NLRP3 pathway protein. Results Compared with the S group, the neurological deficit score, infarct volume, IL-1β, IL-18 contents, apoptosis rate of nerve cells, GasderminD-N (GSDMD-N), Caspase-1, HIF-1α and NLRP3 protein levels were increased in the M group (P<0.05). Compared with the M group, the neurological defect score, infarct volume, IL-1β, IL-18 contents, neuronal apoptosis rate, GSDMD-N, Caspase-1, HIF-1α and NLRP3 protein levels were decreased in the low, medium and high dose Yiqi Shengqing recipe groups (P<0.05). DMOG attenuated the improvement effect of high-dose Yiqi Shengqing recipe on neuronal death in ischemic stroke rats. Conclusion Yiqi Shengqing recipe may reduce the neuronal pyroptosis in ischemic stroke rats by down-regulating HIF-1α/NLRP3 signal pathway.

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Effect of miR-582-5p targeting regulation of FOXO1 on neuronal damage in neonatal rats with hypoxic ischemic encephalopathy

YAN Haifeng, WU Xiaohong, LIN Yuqing, HUO Kaiming, WANG Yingying

2024, 52 (4): 356-361. doi: 10.11958/20230591

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Objective To investigate the effect of microRNA-582-5p (miR-582-5p) on neuronal damage in neonatal rats with hypoxic ischemic encephalopathy (HIE) through targeted regulation of forkhead box transcription factor O1 (FOXO1). Methods Newborn rats (n=90) were randomly grouped into the control (NC) group, the model (HIE) group, the miRNA non-specific control (LV-miRNA-NC) group, the miR-582-5p overexpression (LV-miR-582-5p) group, the miR-582-5p overexpression+mRNA control (LV-miR-582-5p+LV-NC) group and the miR-582-5p overexpression+FOXO1 overexpression (LV-miR-582-5p+LV-FOXO1) group. HIE model was established in all groups except the NC group, and neurological deficits were scored on rats. TTC staining was applied to measure the volume of cerebral infarction. Real-time PCR was applied to detect miR-582-5p and FOXO1 expression. Dual fluorescence reporter gene experiment was applied to detect the targeting relationship between miR-582-5p and FOXO1. HE staining method was applied to observe pathological changes in hippocampal tissue. TUNEL and NeuN fluorescence dual labeling co localization were applied to detect neuronal apoptosis in hippocampal tissue. Immunohistochemistry was applied to detect expressions of FOXO1 and Caspase-3 proteins. Results There was a targeting relationship between MiR-582-5p and FOXO1. Compared with the NC group, the neurological deficit score, cerebral infarction volume, FOXO1 expression, neuronal apoptosis rate, FOXO1 and Caspase-3 protein expression were increased in the HIE group, and the miR-582-5p expression obviously decreased, the hippocampal tissue showed obvious pathological damage (P<0.05). Compared with the LV-miRNA-NC group, the neurological deficit score, cerebral infarction volume, FOXO1 expression, neuronal apoptosis rate, FOXO1 and Caspase-3 protein expression obviously decreased in the LV-miR-582-5p group, and the miR-582-5p expression obviously increased, the pathological damage of hippocampus tissue was obviously improved (P<0.05). LV-FOXO1 was able to reverse the protective effect of LV-miR-582-5p on neuronal damage in HIE rats. Conclusion MiR-582-5p can directly target FOXO1, negatively regulate FOXO1 expression, reduce neuronal apoptosis in HIE neonatal rats, and have a protective effect on neural injury.

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The anti-tumor effect of Runfei Yishen drink on lung cancer bearing rats and its impact on tumor immune microenvironment

ZHANG Caidie, JIN Yan, ZHANG Dede

2024, 52 (4): 362-366. doi: 10.11958/20230694

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Objective To explore the anti-tumor effect of Runfei Yishen drink on lung cancer bearing rats and its impact on tumor immune microenvironment. Methods A lung cancer rat model was established by injecting Lewis lung cancer cells subcutaneously into right quarter rib of rats, and model rats were randomly divided into the model group, the low dose Runfei Yishen drink (Runfei Yishen drink-L) group (3.5 g/kg), the medium dose Runfei Yishen drink (Runfei Yishen drink-M) group (7.0 g/kg), the high dose Runfei Yishen drink (Runfei Yishen drink-H) group (14.0 g/kg) and the positive drug group (6 mg/kg cisplatin). Normal rats were used as the control group. After the intervention, lung index, spleen index and tumor inhibition rate of rats were calculated in each group. Blood samples were collected, and serum levels of transforming growth factor-β (TGF-β), interleukin (IL)-2 and interferon γ (IFN-γ) were detected by ELISA. T lymphocyte subsets in peripheral blood were detected by flow cytometry. Lung tumor tissue was isolated and its pathological changes were detected by HE staining. The expression levels of programmed death receptor 1 (PD-L1) and B-lymphoblastoma-2 (Bcl-2) were detected by Western blot assay. Results Compared with the control group, the pathological injury of the model group was serious, the lung index, TGF-β, CD8⁺, and the expression levels of PD-L1 and Bcl-2 were increased, and the spleen index, IL-2, IFN-γ, CD3⁺, CD4⁺ and CD4⁺/CD8⁺ were decreased (P<0.05). Compared with the model group, the pathological damage of the Runfei Yishen drink-L group, the Runfei Yishen drink-M group and the Runfei Yishen drink-H group gradually improved, the lung index, tumor weight, CD8⁺, TGF-β and the expression levels of PD-L1 and Bcl-2 were decreased, and spleen index, IL-2, IFN-γ, CD3⁺, CD4⁺ and CD4⁺/CD8⁺ were increased (P<0.05). There were no significant differences in above indexes between the Runfei Yishen drink-H group and the positive drug group (P>0.05). Conclusion Runfei Yishen drink can obviously inhibit tumor growth and improve the tumor immune microenvironment in lung cancer bearing rats.

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Effects of different masticatory pressures on alveolar bone remodeling in the pressure side during orthodontic tooth movement in rats

TIAN Jing, WANG Zilong, XIAO Danna, FAN Xiangfei

2024, 52 (4): 367-371. doi: 10.11958/20230630

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Objective To study the effect of different masticatory pressures on alveolar bone remodeling in rats with orthodontic moving teeth. Methods Forty-five male SD rats aged 8 weeks were randomly divided into 3 groups: the baseline group (n=5), the soft diet group (n=20) and the hard diet group (n=20). Rats in the baseline group were sacrificed at the beginning of the experiment. The experimental rat models of right upper first molar movement were established in the soft diet group and the hard diet group. Left side of the maxillary was used as the control group. Each group was fed with corresponding diet. Five rats were sacrificed in each group on the day 3, day 5, day 7 and day 14 after orthodontic treatment respectively. Micro CT was used to measure the mesial movement distance of molar, bone volume/tissue volume (BV/TV), trabecular separation (Tb.Sp) and trabecular thickness (Tb.Th) of the alveolar bone on the pressure side. The number of osteoclasts in alveolar bone was counted after TRAP staining. In situ hybridization was used to detect the expression of RANKL and OPG. Results The amount of tooth movement was smaller on day 14 in the soft diet group than that of the hard diet group (P<0.05). There were no significant differences in BV/TV, Tb.SP and Tb.Th between the soft diet group and the hard diet group. TRAP staining and in situ hybridization showed that the number of osteoclasts and the RANKL/OPG ratio on day 5 and day 7 were lower in the soft diet group than those of the hard diet group (P<0.05). Conclusion The osteoclast activity in the alveolar bone on the pressure side of the orthodontic tooth is decreased and the tooth movement is inhibited under lower masticatory pressure.

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Impacts of cryptotanshinone on autophagy and apoptosis of chondrocytes in rabbit model of knee osteoarthritis by regulating HIF-1α/BNIP3 signaling pathway

WANG Ke, YE Hanlu

2024, 52 (4): 372-378. doi: 10.11958/20230861

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Objective To investigate the impact of cryptotanshinone on autophagy and apoptosis of chondrocytes in a rabbit knee osteoarthritis (KOA) model by regulating the hypoxia inducible factor-1α (HIF-1α)/BCL2 and adenovirus E1B 19-kDa-interacting protein 3 (BNIP3) signaling pathway. Methods New Zealand rabbits were selected to construct the rabbit KOA model using the improved Videman method. Rabbits were randomly divided into the model group, the empty group, the cryptotanshinone group, the HIF-1α knockdown group and the cryptotanshinone+HIF-1α knockdown group, with 9 rabbits in each group. Another 9 New Zealand rabbits were taken as the control group. After grouping and intervention, Lequesne MG knee joint level assessment method was used to score clinical symptoms of knee joints (local pain, gait, joint activity and joint swelling) of rabbits. HE staining was used to detect the cartilage degeneration of rabbit knee joint cartilage tissue, and modified Mankin's score was performed. TUNEL staining was used to detect the apoptosis of articular cartilage histiocytes of rabbits. Enzyme-linked immunosorbent assay (ELISA) method was used to detect serum levels of inflammatory cytokines interleukin-6 (IL-6), IL-18 and IL-10 in rabbits. Western blot experiment was used to detect the expression of autophagy (LC3, Beclin-1), apoptosis (Bax, Cleaved Caspase-3) and HIF-1α/BNIP3 signaling pathway related proteins in knee cartilage tissue of rabbits. Results Compared with the control group, the knee joint cartilage tissue of rabbits showed obvious degenerative symptoms, the scores of local pain, gait, joint activity and joint swelling, the modified Mankin's score, apoptosis rate, levels of serum IL-18 and IL-6, and the expression of LC3Ⅱ/LC3Ⅰ and Beclin-1, Bax, Cleaved Caspase-3, BNIP3 proteins in cartilage tissue were increased in the model group, and the serum level of IL-10 and the expression of HIF-1α protein in cartilage tissue were decreased (P<0.05). Compared with the model group, the symptoms of cartilage tissue degeneration in knee joint of rabbits were alleviated, scores of local pain, gait, joint activity and joint swelling, the modified Mankin's score, apoptosis rate, serum levels of IL-18 and IL-6, and the expression of LC3Ⅱ/LC3Ⅰand Beclin-1, Bax, Cleaved Caspase-3, BNIP3 proteins in cartilage tissue were decreased in the cryptotanshinone group, and the serum level of IL-10 and the expression of HIF-1α protein in cartilage tissue were obviously increased (P<0.05). There was no obvious changes in all indicators of rabbits in the empty group (P>0.05). Compared with the cryptotanshinone group, the symptoms of cartilage tissue degeneration in knee joint of rabbits worsened in the cryptotanshinone+HIF-1α knockdown group, and scores of local pain, gait, joint activity and joint swelling, the modified Mankin's score, apoptosis rate, serum levels of IL-18 and IL-6 and the expression of LC3Ⅱ/LC3Ⅰand Beclin-1, Bax, Cleaved Caspase-3, BNIP3 proteins in cartilage tissue were increased, the level of serum IL-10 and the expression of HIF-1α protein in cartilage tissue were decreased (P<0.05). Changes were opposite to those of the HIF-1α knock-down group. Conclusion Cryptotanshinone can up-regulate HIF-1α to down-regulate BNIP3 expression, inhibit inflammation and autophagy, reduce the degeneration of knee joint cartilage tissue in KOA rabbits, and improve its clinical symptoms.

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Study on the therapeutic effect of andrographolide on psoriasis mouse model by regulating cGAS-STING signal pathway

HE Yanan, CAI Xiang, QIU Baiyi, SUN Bangmei, LI Linghua

2024, 52 (4): 379-386. doi: 10.11958/20230914

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Objective To explore the therapeutic mechanism of andrographolide (Andro) on psoriasis mice by regulating GMP-AMP synthase-stimulator of interferon genes signal pathway (cGAS STING signal pathway). Methods Ninety BALB/c mice were divided into the control group, the model group, the Andro low dose group, the Andro medium dose group, the Andro high dose groups (Andro-L, M, H groups, 10, 30 and 50 mg•kg^-1•d^-1Andro) and the high dose Andro+STING activator DMXAA group (Andro-H+DMXAA group, 50 mg•kg^-1•d^-1 Andro+5 mg•kg^-1•d^-1 DMXAA). Except for the control group, mice of the other groups were administered imiquimod (IMQ) on back to establish the psoriasis mouse model. The psoriasis area of mice was observed, and severity index (PASI) scoring was performed. Serum levels of interleukin-6 (IL-6), tumor necrosis factor α (TNF-α), interleukin (IL)-1β, IL-23, IL-17A, interferon (IFN)-γ and IFN-β were detected by enzyme-linked immunosorbent assay (ELISA). Hematoxylin eosin (HE) staining and toluidine blue staining were used to detect the epidermal thickness and mast cell count. Immunofluorescence staining was applied to detect the expression rates of Ki-67 and CD4 positive cells. RT-qPCR was applied to detect the expression level of cGAS and STING mRNA, and Western blot assay was applied to detect levels of cGAS, STING, interferon regulatory factor-3 (IRF3) and p-IRF3 proteins. Results Compared with the control group, the back skin of mice in the model group showed severe erythema, scales and a large number of inflammatory cell infiltration. The scratch times, PASI score, epidermal thickness, number of mast cells, levels of IL-6, IL-1β, TNF-α, IL-23, IL-17A, IFN-γ and IFN-β, the expression rates of Ki-67 and CD4 positive cell, the expression level of cGAS and STING mRNA and protein, and IRF3 phosphorylation level were increased obviously (P<0.05). Compared with the model group, skin erythema, scales and inflammatory cell infiltration were reduced in the Andro-L, Andro-M and Andro-H groups. The scratch times, PASI score, epidermal thickness, number of mast cells, levels of IL-6, IL-1β, TNF-α, IL-23, IL-17A, IFN-γ and IFN-β, the expression rates of Ki-67 and CD4 positive cells, the expression levels of cGAS and STING mRNA and protein, and IRF3 phosphorylation level were decreased in a dose-dependent manner (P<0.05). STING activator DMXAA reversed the protective effect of Andro on psoriasis mice (P<0.05). Conclusion Andrographolide can improve psoriasis symptoms in mice by inhibiting the cGAS-STING signaling pathway.

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Effect of dexmedetomidine on shivering and hemodynamics during spinal anesthesia in pregnant and postpartum women

ZHU Yongyi, WEN Bolun, WU Qinglin, LIANG Shaoling, ZONG Yindong

2024, 52 (4): 387-390. doi: 10.11958/20231152

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Objective To explore the effect of dexmedetomidine on shivering and hemodynamics during spinal anesthesia in pregnant and postpartum women. Methods A total of 384 cases of cesarean section women with shivering during operation admitted to the Fifth Affiliated Hospital of Guangzhou Medical University were selected as the study subjects and randomly divided into the observation group (n=192) and the control group (n=192). The control group was treated with pethidine for chills, while the observation group was treated with dexmedetomidine. For the anesthesia effect, occurrence of chills, the therapeutic effect of shivering treatment, hemodynamic indicators and serum levels of hypersensitive C-reactive protein (hs-CRP), prostaglandin E2 (PGE2) and cortisol (Cor) were compared between two groups. Results Compared to the control group, the observation group had a shorter stopping time for shivering and better control of shivering (P<0.05). Both groups showed changes in hemodynamic indicators such as diastolic blood pressure (DBP), systolic blood pressure (SBP), heart rate (HR) and oxygen saturation (SpO₂), but the fluctuation amplitude of the observation group was significantly smaller than that of the control group (P<0.05). Stress indicators increased in both groups (P<0.05), but levels of hs-CRP, PGE2 and Cor showed a smaller increase in the observation group compared to those of the control group (P<0.05). The total incidence of adverse reactions was lower in the observation group than that in the control group (P<0.05). Conclusion The clinical application of dexmedetomidine for spinal anesthesia has a good effect in the treatment of shivering in cesarean section women, which can stabilize the hemodynamic level of body and has high clinical application value.

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Changes and clinical significance of white blood cell and platelet parameters of peripheral blood in Brucellosis patients

QIAO Kunyan, LIU Weina, ZHENG Xiaoya, SU Rui, YANG Yuejie

2024, 52 (4): 391-396. doi: 10.11958/20230779

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Objective To study changes of white blood cell (WBC) and platelet parameters in peripheral blood of Brucellosis patients so as to explore the clinical significance. Methods A total of 90 patients with confirmed Brucellosis were included, including 59 in the acute stage and 31 in the chronic stage. In addition, 40 healthy people during the same period were selected as the healthy control group. Basic data such as clinical symptoms were collected. Total WBC count, lymphocyte count, neutrophil count, monocyte count, platelet count, mean platelet volume (MPV) and platelet distribution width (PDW) in peripheral blood were detected by automatic blood analyzer. The counts of CD3⁺T, CD4⁺T and CD8⁺T cells in peripheral blood were detected by flow cytometry. Neutrophil-lymphocyte ratio (NLR) and platelet/lymphocyte ratio (PLR) of patients were calculated. Changes of each cell parameter were analyzed and the clinical diagnostic value of each parameter for Brucellosis were analyzed. Results The proportion of arthralgia and spondylitis were higher in the chronic stage group than those in the acute stage group (P<0.05). Compared with the healthy control group, the total number of WBC, neutrophil count, platelet count and NLR were decreased in the acute stage group, and MPV and PDW were decreased in the chronic stage group, and CD3⁺T cell count and monocyte count were increased (P<0.05). White blood count, lymphocyte count, CD3⁺T cell count, and CD8⁺T cell count were lower in patients with acute Brucellosis with arthritic symptoms than those of patients without arthritis, while PLR was increased (P<0.05). Receiver operating characteristic (ROC) curve analysis showed that the area under the curve of neutrophil count and NLR level for acute Brucellosis was 0.780 and 0.717, respectively, while the area under the ROC curve of CD3⁺T cell count for chronic Brucellosis was 0.748. In acute stage patients, the area under ROC curve of CD3⁺T, CD8⁺T cell count and PLR level for the diagnosis of arthritis was 0.779, 0.768 and 0.722, respectively. Conclusion The indexes of WBC and platelet of peripheral blood in patients with acute and chronic human Brucellosis are changed abnormally, which has a certain reference value in diagnosis of human Brucellosis and arthritis in acute Brucellosis patients.

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Effect of induction chemotherapy on immune function and inflammatory indicators in patients with nasopharyngeal carcinoma

WU Qian, WANG Yi, CHEN Nian, ZHOU Kai, TIAN Xin, XU Hui, GOU Xiaoxia

2024, 52 (4): 397-402. doi: 10.11958/20230792

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Objective To investigate the effect of induction chemotherapy (IC) on immune function and inflammatory markers in patients with stage Ⅲ-Ⅳ nasopharyngeal carcinoma (NPC). Methods A total of 102 patients with stage Ⅲ-Ⅳ NPC confirmed by pathology were selected to receive TPF regimen (docetaxel, cisplatin and 5-fluorouracil, 72 cases) or GP regimen (gemcitabine and cisplatin, 30 cases). The short-term clinical outcomes of patients were evaluated. The immune function of patients with different clinical characteristics was compared before and after IC treatment. T lymphocyte subsets (CD3⁺, CD4⁺, CD8⁺, CD4⁺/CD8⁺), B lymphocyte subsets (CD19⁺) and NK cell percentage in peripheral blood before and after IC were compared. Inflammatory markers interleukin-6, tumor necrosis factor α (TNF-α), neutrophil/lymphocyte ratio (NLR), platelet/lymphocyte ratio (PLR), lymphocyte/monocyte ratio (LMR), and systemic immunoinflammatory index (SII), and changes of total protein (TP), albumin (ALB) and prealbumin (PA) were also compared. The immune function, inflammatory index and nutritional status of patients with different IC regimen and IC cycle number were compared. Results All 102 patients completed IC, of which 0 patient achieved complete response (CR), 87 patients (85.3%) achieved partial response (PR), 13 patients (12.7%) achieved stable response (SD), 2 patients (2.0%) developed PD. The objective response rate (ORR) was 85.3%, and the disease control rate (DCR) was 98.0%. The proportion of CD19⁺ lymphocytes and NK cells decreased after IC, while the proportion of CD3⁺, CD4⁺ and CD8⁺ lymphocytes increased after IC (P<0.05). Levels of NLR, SII, TP, ALB and PA were decreased in NPC patients after IC (P<0.05). After IC, the CD4⁺/CD8⁺ ratio was higher in the GP group than that in the TPF group, and LMR decreased (P<0.05). There were no significant differences in the immune function, inflammatory indexes and nutritional status before and after IC between the ≤3 cycle group and the > 3 cycle group. Conclusion IC is an effective therapy with high disease control rates for NPC patients, and can improve their cellular immune function, decrease humoral immune function, inflammatory indices and nutritional status.

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The correlation between expression of lncRNA MIR503HG and Wnt1 in non-small cell lung cancer tissue and patient survival within 5 years after surgery

QIN Aiying, CHEN Jing, SHAN Fengxiao, LU Hanjie, WU Yang

2024, 52 (4): 403-408. doi: 10.11958/20230852

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Objective To explore the relationship between expression levels of long non-coding RNA (lncRNA) miR503 host gene (MIR503HG) and Wnt1 in non small cell lung cancer (NSCLC) tissue and the survival of patients within 5 years after surgery. Methods A total of 108 NSCLC patients were selected for this study, and lung cancer tissue and paracancer tissue of patients were collected during operation. Fluorescence quantitative PCR was used to detect lncRNA MIR503HG and Wnt1 mRNA expression levels in lung cancer tissue and paracancer tissue. The expression levels of Wnt1 protein in lung cancer tissue and paracancer tissue were determined by immunohistochemistry. Patients with NSCLC were follow up for 5 years after surgery, and their survival status was recorded during the follow-up period. The positive expression of lncRNA MIR503HG and Wnt1 mRNA and protein in paracancer tissue and lung cancer tissue were compared. The expression levels of the above two in lung cancer tissue of NSCLC patients with different outcomes and their differences in different clinicopathological characteristics were compared. Pearson method was used to analyze the correlation between the two expression levels in lung cancer tissue. Kaplan-Meier survival curve was used to analyze the relationship between the expression levels of both in lung cancer tissue and the survival of patients within 5 years after operation. Multivariate Cox regression analysis was used to analyze factors affecting the survival in patients with NSCLC within 5 years after surgery. Receiver operating characteristics (ROC) curves were used to evaluate the predictive values of lncRNA MIR503HG and Wnt1 mRNA expression levels in lung cancer tissue in patients with 5-year postoperative survival. Results LncRNA MIR503HG expression level in lung cancer tissue was lower than those in paracancer tissue. Wnt1 mRNA expression level and Wnt1 protein positive expression rate were higher in lung cancer tissue than that in paracancer tissue (P<0.05). The expression level of lncRNA MIR503HG in lung cancer tissue of NSCLC patients with low differentiation, stage Ⅲ and lymph node metastasis was lower than that of NSCLC patients with medium-high differentiation, stage I-Ⅱ and no lymph node metastasis. The expression level of Wnt1 mRNA in lung cancer tissue of NSCLC patients with low differentiation, stage Ⅲ and lymph nodemetastasis was higher than that of NSCLC patients with medium-high differentiation, stage I-Ⅱ and no lymph node metastasis, respectively (P<0.05). LncRNA MIR503HG and the Wnt1 mRNA expression levels in lung cancer tissue were negatively correlated (P<0.05). A total of 108 NSCLC patients were followed up for 5 years after surgery, with 48 surviving patients (the survival group) and 60 deaths (the death group). The 5-year cumulative survival rate after surgery was higher in the lncRNA MIR503HG high expression group and the Wnt1 mRNA low expression group than that of the lncRNA MIR503HG low expression group and the Wnt1 mRNA high expression group, respectively (P<0.05). Compared with the survival group, lncRNA MIR503HG expression level of lung cancer tissue was decreased and Wnt1 mRNA expression level was increased in the death group (P<0.05). The low expression of lncRNA MIR503HG, high expression of Wnt1 mRNA, low differentiation, stage Ⅲ TNM and lymph node metastasis in lung cancer tissue were all independent risk factors affecting the survival of patients with NSCLC within 5 years after surgery (P<0.05). The area under curve (AUC) of lncRNA MIR503HG, Wnt1 mRNA and their combined in predicting 5-year survival after surgery in NSCLC patients were 0.823, 0.728 and 0.885, respectively, and the combined forecasting was more efficient (P<0.05). Conclusion Low expression of lncRNA MIR503HG and high expression of Wnt1 in lung cancer tissue of NSCLC patients are closely related to the clinicopathological features and survival of patients within 5 years after surgery, and which have a high predictive value for the survival of patients within 5 years after surgery.

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The detection and clinical significance of microparticles derived from lymphocytes in patients with multiple myeloma

WANG Ningfang, ZHAO Chongshan, LIU Fang, ZHAO Penghao, ZHANG Dongdong, CAI Zhuowen, CAI Fangfang

2024, 52 (4): 409-415. doi: 10.11958/20231100

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Objective To investigate the expression and clinical significance of lymphocyte-derived microvesicles (LMP) in patients with multiple myeloma (MM). Methods A total of 65 newly diagnosed MM patients were used as the initial diagnosis group, and 30 health examination volunteers were selected as the control group. After 4 courses of chemotherapy, 8 patients in the initial diagnosis group died within 3 months, and the remaining 57 patients were included in the post chemotherapy group. Flow cytometry was used to detect the expression of LMP in peripheral blood of three groups, as well as the immune phenotype expression of peripheral blood lymphocyte subsets and bone marrow MM cells in the initial diagnosis group. Values of LMP between different therapeutic groups after chemotherapy were compared. The receiver operating characteristic (ROC) curve was used to determine the cutoff value for predicting death for each LMP. For the high (H) group with LMP ≥ cut-off value and the low (L) group with LMP<cut-off value, Kaplan Meier survival analysis was performed. Variables with a P value<0.05 in Kaplan Meier analysis were included into Cox regression analysis to analyze influencing factors of patient death. The differences in lymphocyte subpopulations and myeloma cell immunophenotypes were compared between different LMP groups. Results The proportions of LMP, CD3⁺LMP and CD3⁺CD8⁺LMP were lower in the initial diagnosis group than those in the control group, while the proportions of NKLMP and CD4⁺/CD8⁺LMP were higher than those in the control group (P<0.05). The proportion of CD3⁺CD8⁺LMP was higher in the post chemotherapy group than that in the initial diagnosis group, while the proportion of CD3⁺CD4⁺LMP and the ratio of CD4⁺/CD8⁺LMP were lower than those in the initial diagnosis group (P<0.05). The proportion of CD3⁺LMP and ratio of CD3⁺CD8⁺LMP were higher in the CR+VGPR group than those in the PR+MR+PD group (P<0.01), while the proportion of NKLMP and ratio of CD4⁺/CD8⁺LMP were lower than those in the PR+MR+PD group (P<0.05). Kaplan Meier analysis showed that the median survival time (OS) was shorter in the LLMP group than that of the HLMP group (P<0.01). The median OS was shorter in the LNKTLMP group than that in the HNKTLMP group (P<0.05). Cox regression analysis showed that LLMP and LNKTLMP were independent risk factors for patient mortality, with HR 4.620 and 2.706 (P<0.05). The proportions of CD3⁺CD4⁺T and CD4⁺/CD8⁺T were higher in the LLMP group than those in the HLMP group (P<0.05). The proportion of CD117⁺ in MM cells was higher in the LLMP group than that in the HLMP group (P<0.05). Conclusion Patients with MM have abnormal secretion of LMP. The LMP and NKTLMP are closely correlated with prognosis of MM. Targeted regulation of LMP secretion may improve the survival and prognosis of MM.

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The predictive value of laboratory frailty index for the risk of sepsis and septic shock in elderly CAP inpatients

REN Yan, CHEN Shanping, ZHOU Lihua, WANG Lingxiao, GUAN Lijuan, YANG Yongxue

2024, 52 (4): 416-421. doi: 10.11958/20230833

Abstract ( 309 )

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Objective To investigate the value of laboratory frailty index (FI-LAB) in predicting the risk of sepsis and septic shock in elderly hospitalized patients with community-acquired pneumonia (CAP). Methods A total of 502 elderly CAP patients hospitalized in Chengdu Fifth People's Hospital were retrospectively included. Forty routine laboratory indexes and vital signs collected for the first time within 24 hours after admission were selected to construct FI-LAB. Patients with FI-LAB ≥ 0.35 were defined as the frail group, while patients with FI-LAB< 0.35 were defined as the non-frail group. The occurrence of sepsis and septic shock was observed. The predictive model was established by modified Poisson regression, and the predictive value was analyzed by the ROC curve of the subjects. Results Multivariate modified Poisson regression analysis showed that frailty was a risk factor for sepsis (P<0.001) and septic shock (P=0.001) in elderly patients with CAP (P<0.001). When FI-LAB was multiplied by 100 and included in the multivariate Poisson regression analysis, it was found that for every 0.01 increase in FI-LAB, the risk of sepsis and septic shock increased by 5.9% and 6.2% (both P<0.001). The area under ROC curve of FI-LAB for predicting sepsis and septic shock was 0.865 and 0.850 (both P<0.001), respectively, and the best cut-off values were 0.28 and 0.42, respectively. Conclusion Frailty increases the risk of CAP complications with sepsis and septic shock. FI-LAB can predict the risk of sepsis in CAP patients and detect high-risk groups early.

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Correlation between serum miR-34a and miR-182 levels before PCI and postoperative contrast-induced nephropathy occurrence in elderly patients with acute coronary syndrome

XUE Wenping, QIN Wei, LIU Tingting, ZHANG Aiwen, SHI Fei

2024, 52 (4): 422-426. doi: 10.11958/20231183

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Objective To investigate the relationship between serum microRNA (miR)-34a and miR-182 levels before percutaneous coronary intervention (PCI) and postoperative contrast-induced nephropathy (CIN) occurrence in elderly patients with acute coronary syndrome (ACS). Methods A total of 146 elderly patients with ACS who underwent PCI were included. Clinical data of ACS patients were collected. The indexes of preoperative blood lipid and renal function were detected by automatic biochemical analyzer. Serum levels of miR-34a and miR-182 were determined by real time fluorescence quantitative PCR. Patients were divided into the CIN group (20 cases) and non-CIN group (126 cases) according to whether CIN occurred after PCI. Clinical data, preoperative blood lipids, renal function indexes, serum miR-34a, miR-182, postoperative serum creatinine (Scr) and glomerular filtration rate (eGFR) were compared between the CIN group and the non-CIN group. The correlation of preoperative serum miR-34a, miR-182, postoperative Scr, and eGFR in elderly patients with ACS were analyzed, and factors affecting the occurrence of CIN in elderly patients with ACS after PCI were also analyzed. Receiver operating characteristic (ROC) curve was used to evaluate the predictive value of preoperative serum miR-34a and miR-182 levels for occurrence of CIN in elderly patients with ACS after PCI. Results Preoperative serum miR-34a and miR-182 levels and postoperative Scr levels were higher in the CIN group than those in the non-CIN group, while postoperative eGFR level was lower than those in the non-CIN group (P<0.05). Preoperative serum miR-34a and miR-182 were positively correlated with postoperative Scr, and negatively correlated with postoperative eGFR in elderly patients with ACS (P<0.05). Preoperative serum miR-34a was positively correlated with miR-182 (P<0.05). The increase of serum miR-34a and miR-182 levels before surgery were an independent risk factor for occurrence of CIN in elderly patients with ACS after PCI (P<0.05). The area under curve (AUC) of preoperative serum miR-34a, miR-182 and their combination in predicting postoperative occurrence of CIN in elderly ACS patients after PCI were 0.881, 0.888, and 0.964, respectively. The AUC predicted by the two combined was higher than that predicted by each separately (P<0.05). Conclusion The increased serum levels of miR-34a and miR-182 before surgery are risk factors for occurrence of CIN in elderly patients with ACS after PCI, and the combination of the two can effectively predict the occurrence of CIN.

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Analysis of influencing factors of chronic obstructive pulmonary disease in permanent residents in some area of Tianjin

FENG Liting, LI Li, XIE Xin, WANG Xing

2024, 52 (4): 427-431. doi: 10.11958/20230994

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Objective To analyze the pulmonary ventilation function and influencing factors in residents in some area of Tianjin and provide a basis for the precise prevention and control of chronic obstructive pulmonary disease (COPD) in Tianjin. Methods A total of 5693 permanent residents aged 35 years and above in Heping District, Jinnan District and Baodi District of Tianjin were randomly selected from June to December 2022. The basic information, smoking history, treatment and China COPD Screening Questionnaire (COPD-SQ) were collected. According to the COPD-SQ result, the subjects were divided into the COPD low-risk group (COPD-SQ score<16 points, 4 648 cases) and the COPD high-risk group (COPD-SQ score>16 points, 1 045 cases). The physical examination and the pulmonary ventilation function testing were completed. Multivariate Logistic regression was used to analyze independent risk factors for COPD disease. Results The included subjects with the body quality index 25.1 (23.0, 27.6) kg/m² were 56 (48, 64) years old, including 2 393 (42.0%) men, 1 381 (24.3%) smokers and 650 (11.4%) proposed COPD. Only 18 patients were received standard treatment, accounting for 2.8% of the patients with suspected COPD. Compared with the COPD low-risk group, the proportion of males and smokers, age, waist-to-hip ratio, blood pressure and proposed COPD were higher in the COPD high-risk group. BMI, peripheral percutaneous oxygen saturation, FVC percentage of predicted value, FEV1 percentage of predicted value, FEV1 percentage of FVC, PEF percentage of predicted value, MMEF percentage of predicted value, and PEF50 percentage of predicted value were lower (all P<0.05). The proportion of smokers and proposed COPD in Heping District, Jinnan District and Baodi District had statistical significance (P<0.05). The multivariate Logistic regression analysis showed that independent risk factors of COPD morbidity included gender, smoking and age (P<0.05). Conclusion The smoking rate of residents aged 35 and above in some area of Tianjin is high. Age, smoking status and gender are associated with the incidence of COPD.

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Pathogenic Surveillance of HFMD and molecular epidemic characteristics of major epidemic types in Shaanxi province, China, from 2020 to 2022

DU Yanhua, GUAN Luyuan, FENG Xuan, ZHENG Yuan, YU Pengbo

2024, 52 (4): 432-437. doi: 10.11958/20231032

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Objective To investigate the composition and prevalence of hand-foot-mouth disease (HFMD) in Shaanxi province from 2020 to 2022. Methods Nucleic acid detection results and case data of HFMD surveillance cases in cities of Shaanxi province were collected. Positive samples of enterovirus nucleic acid detection were isolated. PCR amplification, sequence determination and gene subtype identification of enterovirus VP1 region of epidemic dominant pathogen were performed. Results From 2020 to 2022, a total of 6 352 samples of HFMD cases were collected in Shaanxi province, and 4 417 samples tested positive for enterovirus nucleic acid, with a positive rate of 69.54%. In 2020 and 2021, other enteroviruses excluding EV-A71 and EV-A16 were the main epidemic types, with a composition ratio of 95.43% and 74.35%, respectively. In 2022, CV-A16 was the main epidemic type, with a composition ratio of 52.88%. CV-A6 and CV-A10 types of other enteroviruses were detected in 4 145 laboratory confirmed cases of HFMD in 8 cities. CV-A6 accounted for 75.31%, 62.56% and 61.15% of other enteroviruses, which was the dominant epidemic type of other enteroviruses in Shaanxi province. From 2020 to 2022, 239 isolates, 151 CV-A16 isolates, 72 CV-A6 isolates, 6 CV-A10 isolates, 2 CV-A4 isolates and 8 CV-B3 isolates were sequenced in Shaanxi province. CV-A16 and CV-A6 were the main epidemic types. According to phylogenetic analysis, the D3 subtype was the main epidemic subtype of CV-A6, while B1a and B1b coexisted for CV-A16. The proportion of B1a subtype increased year by year in 2021 and 2022, and it replaced B1b as the main epidemic subtype. Conclusion From 2020 to 2022, the prevalence of HFMD in Shaanxi province is at a lower level than that in previous years, and the main pathogens are other enteroviruses and CVA16.

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Research progress on the role and mechanism of 5-hydroxytryptamine in gastrointestinal diseases

SHAN Zihong, HU Xiaoqing, LI Feng, HUANG Yongkun

2024, 52 (4): 438-442. doi: 10.11958/20231309

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5-hydroxytryptamine (5-HT) is an important neurotransmitter in brain-gut axis pathway, and its secretion is closely related to intestinal flora, intestinal immunity and intestinal motility. Abnormal metabolic pathway of 5-HT is involved in the occurrence and development of gastrointestinal diseases, such as irritable bowel syndrome, inflammatory bowel disease, chronic constipation and functional dyspepsia. This article reviews the mechanism of 5-HT in different gastrointestinal diseases in order to explore new ideas for clinical treatment.

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Research progress of SIRT1 in the pathogenesis of diabetic cardiomyopathy

XIE Youcheng, WANG Fei, XU Jin, YU Xiaohui

2024, 52 (4): 443-448. doi: 10.11958/20231289

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Diabetic cardiomyopathy (DCM) is one of the most serious cardiovascular complications in diabetic patients, and it is also the main cause of increased mortality in diabetic patients, which has become a major global public health problem. As an important intracellular regulatory protein in body, sirtuin 1 (SIRT1) plays an important role in many biological processes, including reducing oxidative stress in cardiomyocytes, maintaining Ca²⁺ homeostasis in myocardial mitochondria, reducing myocardial endoplasmic reticulum stress, improving myocardial mitochondrial dysfunction, and inhibiting the activation of renin-angiotensin-aldosterone system. SIRT1 may be a potential therapeutic target for DCM. Further research targeting SIRT1 can provide a new theoretical basis for the clinical treatment of DCM. This article reviews the specific role of SIRT1 in the pathogenesis and treatment strategies of DCM.

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