Tianjin Medical Journal

Emphasize the role of electrophysiological measurements in the diagnosis of amyotrophic lateral sclerosis

ZHU Ju, ZHANG Zhecheng

2023, 51 (7): 673-676. doi: 10.11958/20230474

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Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease characterized by loss of upper and lower motor neurons, resulting in progressive weakness and wasting of all voluntary muscles, culminating in respiratory failure. Neuroelectrophysiological detection plays an an essential role in early diagnosis of ALS, and can even detect subclinical dysfunction. Nerve conduction and needle electromyography, as standard neurophysiological detection method, are very important for early detection of lower motor neuron disease. Many emerging techniques for estimating the number of motor units have been proposed to increase diagnostic sensitivities and used to assess ALS progression.

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Risk factors of silent aspiration in patients with amyotrophic lateral sclerosis and the predictive value of ALSFRS-R scale

XIA Xiaoqian, ZHANG Wei, CHANG Xueli, GUO Junhong

2023, 51 (7): 677-680. doi: 10.11958/20230473

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Objective To detect risk factors of silent aspiration in patients with amyotrophic lateral sclerosis (ALS) and the diagnostic value of ALSFRS-R scale. Methods A total of 68 patients with ALS were recruited in our study. According to the results of Videofluoroscopic Swallowing Study (VFSS), patients were divided into the silent aspiration group (9 cases) and the non-silent aspiration group (59 cases). The muscle strength of neck and limb was assessed using the Medical Research Council score (MRC) scale, and the independent influencing factors of silent aspiration were assessed using Logistic regression analysis. Receiver operating characteristic (ROC) curve was used to analyze the diagnostic value of ALSFRS-R bulbar subscales and swallowing items for silent aspiration. Results The results of univariate analysis showed that compared with the non-silent aspiration group, cervical flexor muscle strength and right proximal upper limb muscle strength were decreased in the silent aspiration group (P<0.05), and the proportion of patients with bulbar onset was higher in the silent aspiration group (P<0.05). Multivariate Logistic regression analysis showed that patients with limb onset were less likely to have occult aspiration than those of bulbar onset (OR=0.001, 95%CI: 0.000-0.222, P=0.014). The higher the cervical flexor muscle strength was, the less the risk of occult aspiration was (OR=0.089, 95%CI: 0.010-0.814, P=0.032). The results of ROC curve showed that the ALSFRS-R bulbar subscales and swallowing items had limit clinical diagnostic value for ALS patients with silent aspiration, and the areas under the curve (AUC) were 0.774 (95%CI：0.741-0.924) and 0.781 (95%CI： 0.757-0.934), respectively (P<0.05). When the Youden index was the maximum, the optimal cut-off value of ALSFRS-R bulbar subscales was 11, and the sensitivity and specificity for the diagnosis of silent aspiration were 88.89% and 57.63%, respectively. The optimal cut-off value of ALSFRS-R swallowing items was 3, and the sensitivity and specificity for the diagnosis of silent aspiration were 77.78% and 67.80%, respectively. Conclusion For ALS patients with bulbar onset and poor cervical flexor muscle strength, attention should be paid to the screening of dysphagia and early clinical intervention. ALSFRS-R bulbar subscales and swallowing items have limited diagnostic value for silent aspiration, and new scales need to be developed to assess the risk of silent aspiration in ALS patients.

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Clinical characteristics of amyotrophic lateral sclerosis patients

LIU Yufei, ZHU Ju, LIU Na, REN Yanping, SUN Xiaohui, TIAN Li, ZHANG Zhecheng

2023, 51 (7): 681-686. doi: 10.11958/20230447

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Objective To analyze the clinical characteristics and epidemiological status of amyotrophic lateral sclerosis (ALS). Methods A total of 110 ALS patients (62 males and 48 females) with a median age of 60 years of onset, were collected. The basic information, site of onset, diagnostic process, modified amyotrophic lateral sclerosis function score (ALSFRS-R), rate of disease progression (ΔFS), electrophysiology and pulmonary function were analyzed comprehensively. The patients were followed up by outpatient or telephone every 3 to 6 months to record changes of their condition, the application of non-invasive positive pressure ventilation (NIPPV), gastric tube, percutaneous gastrostomy (PEG) and the treatment of riluzole. Results Among 110 patients, limb onset accounted for 90 patients (81.8%). The delay diagnosis was 12 (7, 20) months. The misdiagnosis interval was 11 (7, 17) months, and the misdiagnosis rate was 81.3% (74 cases). The most misdiagnosed cases were cervical spondylosis and lumbar spondylosis. The ALSFRS-R score was 39 (36, 45) points, and the ΔFS was 0.6 (0.3, 1.0) points/month. The proportion of confirmed and suspected diagnosis was higher in patients with ball onset than that in patients with limb onset (90% vs. 83%, χ²=2.098, P<0.05). Compared with patients of rapid disease progression, the age of onset was younger in patients with moderate and slow disease progression. Compared with patients of slow disease progression, patients of rapid and moderate disease progression had shorter diagnostic delay. Results of EMG in 90 (81.8%) patients indicated that lower motor neuron lesion segment was greater than or equal to the clinically involved segment. The first pulmonary function examination indicated that 50% (45 cases) patients had restrictive ventilation dysfunction, and the forced vital capacity (FVC)<70% accounted for 17.8% (16 cases). After discharge, the treatment rate of riluzole was 85.6%, and the application rate of NIPPV and PEG was low. During the follow-up, 50 patients (45.5%) died, and the median survival time was 44.0 months. Median survival time was shorter in patients with onset age ≥60 years than that in patients under 60 years (36 months vs. 50 months, P<0.05). Survival time was shorter in patients with rapid disease progression than that in patients with moderate and slow disease progression (27 months vs. 32 months vs. 65 months, P<0.01). Conclusion A full and correct understanding of clinical features of ALS is helpful to make a reasonable diagnosis and treatment plan for patients and improve the prognosis.

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Clinical observation of CT-guided and endoscopic percutaneous gastrostomy in patients with amyotrophic lateral sclerosis

LIAN Ling, LIN Weiyuan, XIANG Xianhong, YAO Xiaoli

2023, 51 (7): 687-689. doi: 10.11958/20230443

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Objective To compare the clinical effects of CT-guided and endoscopic surgical percutaneous gastrostomy in patients with amyotrophic lateral sclerosis (ALS). Methods The clinical data of 104 patients with ALS were retrospectively analyzed, including 48 patients underwent CT-guided percutaneous gastrostomy (CT-guided group) and 56 patients underwent endoscopic percutaneous gastrostomy (endoscopic group). The disease status, length of hospital stay, postoperative length of hospital stay, postoperative complications, and survival rates of six months and one year after surgery were compared between the two groups. Results There were no significant differences in age, disease course, dysphagia time, height, body weight and preoperative nutritional status between the two groups (P>0.05). The length of hospital stay and postoperative length of stay were shorter in the CT-guided group than those in the endoscopic group (P<0.05). There was no significant difference in the incidence of complications between the endoscopic group (19.6%) and the CT-guided group (8.3%, P>0.05). There was no significant difference in one-year survival rate between the endoscopic group and the CT-guided group (91.0% vs. 93.8%, Log-rank χ²=1.606, P>0.05). Conclusion CT-guided percutaneous gastrostomy shows its advantages in ALS, especially in shortening the length of hospital stay and postoperative length of hospital stay in ALS patients, and is worthy of widespread clinical application.

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Neuroelectrophysiological study of sensory system in amyotrophic lateral sclerosis

REN Yanping, ZHU Ju, TIAN Li, SUN Xiaohui, LIU Yufei, LIU Na, ZHANG Zhecheng

2023, 51 (7): 690-693. doi: 10.11958/20230451

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Objective To evaluate the functional status of the sensory system in patients with amyotrophic lateral sclerosis (ALS) by neuroelectrophysiological techniques. Methods According to the revised El Escorial diagnostic criteria, 66 patients with clinically confirmed and proposed limb-onset ALS were included and divided into the sensory symptom (sALS, n=13) group and without sensory symptom (nsALS, n=53) group according to the presence or absence of clinical sensory symptoms. Sixty healthy individuals were selected as the health control (HC) group. Sensory nerve conduction studies (SNCS) and skin sympathetic response (SSR) of bilateral upper and lower limbs were detected by Keypoint 4 electromyography evoked potential instrument. Pathway pain and sensation assessment system was used to conduct contact heat evoked potential (CHEP) detection of bilateral anterolateral leg and forearm volar side. Results (1) The abnormal rates of SNCS, SSR and CHEP in the ALS group were 0%, 21.2% (14/66) and 27.3% (18/66), respectively. The abnormal rates of SSR and CHEP in the sALS group were 30.8% (4/13) and 38.5% (5/13). The abnormal rates of SSR and CHEP in the nsALS group were 18.9% (10/53) and 24.5% (13/53). (2) Compared with the HC group, the latency of SSR in lower limbs was prolonged in the sALS group and the nsALS group, the latency of N wave in CHEP of upper and lower limbs was prolonged, and the amplitude of N-P wave was decreased (P<0.05). Compared with the nsALS group, the sALS group showed prolonged N wave latency and reduced N-P wave amplitude for anterolateral calf stimulation by CHEP detection (P<0.05). Conclusion ALS patients have small fiber damage in the sensory system.

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Advances in clinical characteristics and prognostic factors of amyotrophic lateral sclerosis

ZHAO Xiaoquan, LU Zuneng

2023, 51 (7): 694-696. doi: 10.11958/20230475

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Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease with marked clinical heterogeneity, with an extremely variable time course from diagnosis to death or severe disability. The heterogeneity of ALS hinders the accurate prediction of endpoints and the development of novel therapies. Further identification of ALS prognostic factors will help to build a good survival prediction model, accurately predict survival time, and promote the development of ALS treatment.

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Research progress of amyotrophic lateral sclerosis related immune cells

SU Boyang, HE Zhengqing, HUANG Xusheng

2023, 51 (7): 697-700. doi: 10.11958/20230187

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Abstract: Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease, and its pathogenesis includes glutamate excitotoxicity, oxidative stress injury, abnormal aggregation of neurofilaments, abnormal aggregation of intracellular calcium, neurotrophic factor deficiency, mitochondrial dysfunction, RNA metabolism disorder and immune dysfunction. This article reviews the effects of monocytes/macrophages, natural killer cells, astrocytes, microglia, CD4⁺T cells, CD8⁺ T cells and the number and function of regulatory T cells on ALS.

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Study of cynaroside on apoptosis and expression of inflammatory factor in model cells of Alzheimer’s disease

WANG Cui, YANG Chang, JIN Yu, GAO Mi, ZHANG Wen, WANG Qiong, JIN Haitao

2023, 51 (7): 701-706. doi: 10.11958/20221922

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Objective To explore the effect and possible mechanism of cynaroside on apoptosis and expression of inflammatory factor in model cells of Alzheimer's disease (AD) by down-regulating mitogen-activated protein kinase kinase kinase 3 (MEKK3). Methods PC12 cells were cultured in vitro and incubated with different doses (6.25 mg/L, 12.5 mg/L, 25 mg/L) of cynaroside for 24 h to establish an AD cell model induced by Aβ_25-35. In addition, MEKK3 small interfering RNA or overexpression vector was transfected into PC12 cells, and Aβ_25-35 induced AD cell model was established after incubation with 25 mg/L luteolin for 24 h. CCK-8 was used to detect inhibition rate of cell proliferation, and flow cytometry was used to detect apoptosis rate. ELISA was used to detect inflammatory factors (TNF-α, IL-6, IL-1β). The protein expression of MEKK3 was detected by Western blot assay. Results Cynaroside could decrease the proliferation inhibition rate, apoptosis rate, inflammatory cytokines (TNF-α, IL-6, IL-1β) and MEKK3 protein expression in PC12 cells induced by Aβ_25-35 (P<0.05). Knockdown of MEKK3 could reduce the proliferation inhibition rate, apoptosis rate and inflammatory cytokines (TNF-α, IL-6, IL-1β) of Aβ_25-35-induced PC12 cells, while overexpression of MEKK3 showed the opposite effect. Upregulation of MEKK3 expression attenuated effects of cynaroside on the proliferation inhibition rate, apoptosis rate and inflammatory factors of Aβ_25-35-induced PC12 cells. Conclusion Cynaroside may inhibit apoptosis and expression of inflammatory factors of AD model cells by down-regulating MEKK3, which has potential value in the treatment of AD.

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Effects of taurine on proliferation, apoptosis and migration of human pancreatic cancer cell lines BxPC-3 and PANC-1

GAO Huijie, LI Qian, TIAN Bin, ZHU Riming, LIU Chao

2023, 51 (7): 707-712. doi: 10.11958/20221545

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Objective To investigate the effects of taurine (Tau) on the proliferation, apoptosis and migration of pancreatic cancer ductal cells. Methods BxPC-3 and PANC-1 cells were cultured in vitro and pretreated with different concentrations of Tau (0, 10, 20, 40, 80, 160 mmol/L). Cell counting kit-8 (CCK-8), cell scratch assay and Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) method were used to observe the effect of Tau on the proliferation, migration and apoptosis of BxPC-3 and PANC-1 cells. Quantitative real-time polymerase chain reaction (qPCR) and Western blot assay were used to detect the effect of Tau on mRNA and protein expression of relevant apoptosis and cell cycle molecules in BxPC-3 and PANC-1 cells. Results Tau significantly inhibited the proliferation and migration activities of BxPC-3 and PANC-1, and Tau could promote the apoptosis of BxPC-3 and PANC-1 cells. Compared with the control group, expression levels of P53, P21 and BAX in BxPC-3 cells treated with Tau showed a significant upward trend, while the expression of PCNA was significantly decreased. The expression levels of Bcl-2, PCNA, CyclinA2, CyclinB1, CyclinE, CDK1, CDK2, CDK4 and CDK6 in PANC-1 cells were significantly lower than those in the normal control group, while the expression levels of P53 and P21 showed an upward trend. Conclusion Tau can inhibit the proliferation and migration activity of BxPC-3 and PANC-1 pancreatic cancer cells, and promote cell apoptosis, possibly by affecting related apoptosis genes and cyclins.

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Effects of circRASSF2 targeting miR-1343-3p on the proliferation and apoptosis of breast cancer MDA-MB-231 cells

ZHAO Jinjin, WANG Zhenyu, MA Zhenxiu

2023, 51 (7): 713-717. doi: 10.11958/20221254

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Objective To explore the effect of circRASSF2 on the proliferation and apoptosis of breast cancer MDA-MB-231 cells and its molecular mechanism. Methods Real-time fluorescence quantitative polymerase chain reaction (RT-qPCR) was used to detect circRASSF2 and miR-1343-3p expression levels in 37 breast cancer tissue samples and adjacent tissue samples. Breast cancer MDA-MB-231 cells were divided into the si-circRASSF2 group, the si-NC group, the miR-1343-3p group, the miR-NC group, the si-circRASSF2+anti-miR-1343-3p group and the si-circRASSF2+anti-miR-NC group. Tetramethylazolium salt colorimetry method (MTT) was used to detect cell viability. Clone formation experiment was used to detect the number of cell clones. Annexin V-FITC/PI double staining method was used to detect cell apoptosis. The relative expression levels of Cleaved-caspase 3 and Cleaved-caspase 9 proteins were detected by Western blot assay. Dual luciferase reporter experiment was used to detect targeting relationship between circRASSF2 and miR-1343-3p. Results Compared with paracancer tissue samples, the expression level of circRASSF2 was increased and the expression level of miR-1343-3p was decreased in breast cancer tissue samples (P<0.01). Compared with the si-NC group, the OD value of MDA-MB-231 cells was decreased in the si-circRASSF2 group, the number of cell clone formation was decreased, and the apoptosis rate was increased (P<0.05). Compared with the miR-NC group, the OD value of MDA-MB-231 cells was decreased in the miR-1343-3p group, the number of cell clone formation was decreased, and the apoptosis rate was increased (P<0.05). Compared with the miR-NC group, WT-circRASSF2 luciferase activity decreased in the miR-1343-3p group (P<0.05). Compared with the si-circRASSF2+anti-miR-NC group, the OD value and the number of cell clone formation were increased, and the apoptosis rate was decreased in the si-circRASSF2+anti-miR-1343-3p group (P<0.05). Conclusion The inhibition of circRASSF2 expression can up-regulate miR-1343-3p, thus inhibiting the proliferation of breast cancer MDA-MB-231 cells and promoting apoptosis.

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Protective mechanism of catalpol on IL-1β-induced chondrocyte injury

MA Ling, ZHONG Liguo, CUI Yuru, LIU Bin

2023, 51 (7): 718-723. doi: 10.11958/20221441

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Objective To investigate the effect of catalpol on interleukin-1β (IL-1β)-induced chondrocyte injury and its possible mechanism. Methods Human knee joint chondrocytes were isolated, cultured and divided into the control (Con) group, the IL-1β group (treated with 10 μg/L IL-1β for 48 h), the IL-1β+catalpol-low group, the IL-1β+catalpol -medium group, the IL-1β+catalpol -high group (treated with 10, 20 and 50 ng/L catalpol and then treated with 10 μg/L IL-1β for 24 h), the IL-1β+miR-NC group (transfected with miR-NC, treated with 10 μg/L IL-1β for 24 h), the IL-1β+miR-140-5p group (transfected with miR-140-5p mimics, treated with 10 μg/L IL-1β for 24 h), the IL-1β+catalpol+anti-miR-NC group (after transfection with anti-miR-NC, treated with 50 ng/L catalpol and 10 μg/L IL-1β for 24 h) and the IL-1β+catalpol+anti-miR-140-5p group (transfected with anti-miR-140-5p, treated with 50 ng/L catalpol and 10 μg/L IL-1β for 24 h). Cell viability was detected by cell count Kit 8 (CCK-8 method). Levels of interleukin-6 (IL-6), tumor necrosis factor α (TNF-α) and interferon γ (IFN-γ) were detected by enzyme-linked immunosorbent assay (ELISA). The apoptosis rate was detected by flow cytometry. The expression of miR-140-5p was detected by real-time quantitative fluorescence PCR (qPCR). Expression levels of cleaved aspartate specific cysteine proteinase (Cleaved-caspase) 3 and Cleaved-caspase 9 were detected by Western blot assay. Results Compared with the IL-1β group, levels of IL-6, TNF-α, IFN-γ, cell apoptosis rate, Cleaved-caspase 3 and Cleaved-caspase 9 protein expression levels were decreased in the IL-1β+catalpol-low group, the IL-1β+catalpol-medium group and the IL-1β+catalpol-high group, but cell viability and miR-140-5p expression were increased (P<0.05). Compared with the IL-1β+miR-NC group, levels of IL-6, TNF-α, IFN-γ, cell apoptosis rate, Cleaved-caspase 3 and Cleaved-caspase 9 protein expression levels were decreased in the IL-1β+miR-140-5p group (P<0.05), and cell viability and miR-140-5p were increased (P<0.05). Compared with the IL-1β+catalpol +anti-miR-NC group, levels of IL-6, TNF-α, IFN-γ, cell apoptosis rate, Cleaved-caspase 3 and Cleaved-caspase 9 protein expression levels were increased in the IL-1β+ catalpol+anti-miR-140-5p group (P<0.05), and cell viability was decreased (P<0.05). Conclusion Catalpol may reduce IL-1β-induced chondrocyte injury by up-regulating the expression of miR-140-5p, inhibiting the release of cellular inflammatory factors and apoptosis.

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Moderately elevating blood potassium concentration can improve mitochondrial function of brain tissue in cardiopulmonary resuscitation rats

SHI Jiaxin, LI Nuo, YANG Yegui, FANG Wei, QIN Sina, HUANG Jingju, CHEN Menghua

2023, 51 (7): 724-728. doi: 10.11958/20221699

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Objective To explore whether moderately hyperkalemia can improve mitochondrial function of brain tissue during cardiopulmonary resuscitation (CPR) in rats. Methods Eighty rats were randomly divided into five groups: the sham (SH) group, the normal saline (NS) group, the low dosage of potassium chloride (LK) group, the mid-dosage of potassium chloride (MK) group and the high-dosage of potassium chloride (HK) group, with 16 rats in each group. Cardiac arrest (CA) /CPR model was established by transesophageal electrical stimulation to induce ventricular fibrillation. Rats were received equal volume of normal saline and the corresponding dose of potassium chloride via intravenous injection when CPR was initiated at 6 min after CA. The mean arterial pressure (MAP) and the blood potassium concentration were detected after return of spontaneous circulation(ROSC). The neurological deficit scores (NDS), the changes of cell morphology of hippocampal tissue, the content of adenosine-triphosphate, the activity of Na⁺/K⁺-ATP enzymes, the respiratory chain complex (Ⅰ/Ⅱ/Ⅲ/Ⅳ) of the hippocampal tissue at 24 h post ROSC were recorded and observed between different groups. Results Compared with the NS group, the blood potassium was significantly higher in the MK group and the HK group at 1 min and 10 min post ROSC (P<0.05). Compared with the NS group, the MAP after ROSC was not significantly affected, and the NDS was significantly increased at 24 h after ROSC in the MK group (P<0.05). Compared with the NS group, the MAP within 8 min after ROSC was significantly decreased (P<0.05), and the NDS was not significantly improved at 24 h post ROSC in the HK group. Compared with the NS group, the activity of hippocampal respiratory chain complex Ⅲ was increased in the LK group, and the content of ATP was increased (P<0.05). Compared with the NS group, the activity of respiratory chain complex (Ⅰ/Ⅱ/Ⅲ), the content of ATP and the activity of Na⁺/K⁺-ATP enzyme at 24 h post ROSC were increased in the MK group (P<0.05), and hippocampal cell morphology was improved 24 h after ROSC. Compared with the NS group, the activity of respiratory chain complex Ⅲ increased in the HK group (P<0.05). The MK group showed the most obvious improvement in each evaluation index. Conclusion Moderately elevating blood potassium at onset of CPR can improve mitochondrial function and alleviate cerebral ischemia -reperfusion injury in CPR.

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Effects of linezolid on bacterial load and bone repair in rats with chronic osteomyelitis induced by MRSA infection

CHEN Lixi, CHEN Yuanliang, ZHUO Zeming, WANG Hejie

2023, 51 (7): 729-733. doi: 10.11958/20221569

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Objective To explore the effects of linezolid on bacterial load and bone repair in rats with chronic osteomyelitis induced by methicillin-resistant Staphylococcus aureus (MRSA) infection. Methods A total of 45 Wistar rats were randomly and averagely divided into the sham operation group, the model group and the linezolid group, 15 rats in each group. Rats in the model group and the linezolid group were inoculated with 10 μL of MRSA (1×10⁸ CFU/mL) after bilateral proximal tibia bone defects to construct rat models of chronic osteomyelitis. At 7 d after modeling, rats in the linezolid group was intraperitoneally injected with linezolid (54 mg/kg), while the sham operation group and the model group were given the same volume of glucose injection for 14 d. The wound healing, scores of Rissing and tibia X-ray Norden, and bacterial load were observed. The pathological changes of tibia tissue were observed by HE staining. The levels of transforming growth factor-β1 (TGF-β1), interleukin (IL)-1β and IL-6 in skeletal muscle tissue around lesions were detected by enzyme-linked immunosorbent assay. The expression levels of TGF-β1, activin receptor-like kinase 1 (ALK-1), Smad1/5, p-Smad1/5 and bone morphogenetic protein 2 (BMP-2) in tibial tissue were detected by Western blot assay. Results All the sham operation group healed in class-A. In the model group, there were 3 rats with class-B healing and 12 rats with class-C healing. In the linezolid group, there were 9 rats with class-B healing and 6 rats with class-C healing, there was a significant difference between the two groups (χ²=35.111, P<0.01). Rissing score, Norden score and bacterial load were lower in the linezolid group than those in the model group (P<0.05). In the model group, there were local bone destruction, intramedullary and subperiosteal abscesses, inflammatory cell infiltration and local fibrosis. In the linezolid group, bone destruction, inflammatory cell infiltration and fibrosis were significantly relieved. Compared with the sham operation group, levels of TGF-β1, IL-1β and IL-6 in skeletal muscle tissue, and expression levels of TGF-β1, ALK-1 and p-Smad1/5 in tibia tissue were increased, while BMP-2 was decreased in the model group (P<0.05). Compared with the model group, linezolid intervention could reverse changes of the above indexes (P<0.05). Conclusion Linezolid can relieve bacterial load and promote bone repair in rats with chronic osteomyelitis induced by MRSA infection by inhibiting the activation of TGF-β/BMP pathways.

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Influence of formononetin on oxidative stress injury in gestational diabetes mellitus rats

TIAN Yajing, YANG Xue, WANG Jing, GE Wenjie, HE Yuling

2023, 51 (7): 734-738. doi: 10.11958/20221450

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Objective To explore the influence of formononetin on oxidative stress injury in gestational diabetes mellitus (GDM) rats by regulating nuclear factor erythroid-2-related factor 2 (Nrf2/hemeoxygenase-1 (HO-1)/NADPH quinone oxidoreductase-1 (NQO1) signaling pathway. Methods SD pregnant rats were injected intraperitoneally with streptozotocin to induce GDM model. Model rats were randomly grouped into the model group, the low-dose formononetin (50 mg/kg) group, the high-dose formononetin (100 mg/kg) group, the ML385 (Nrf2 inhibitor, 20 mg/kg) group, and the high-dose formononetin (100 mg/kg) + ML385 (20 mg/kg) group, 9 rats per group. Another 9 normal pregnant rats were regarded as the control group. After treatment with formononetin and ML385, body weight, serum fasting blood glucose (FSG), total cholesterol (TC), triglyceride (TG) levels, embryo survival rate, fetal body weight, the overall morphology and ultrastructure of placenta, levels of malondialdehyde (MDA), glutathione (GSH), superoxide dismutase (SOD) and total antioxidant capacity (TAC) in serum and placental tissue were detected in rats of each group. The protein expression of Nrf2/HO-1/NQO1 pathway in placental tissue of rats was also detected in each group. Results Compared with the control group, the placental ultrastructure was damaged in the model group, and the embryonic survival rate, serum and placental tissue GSH, SOD, TAC levels, placental tissue Nrf2, HO-1, NQO1 protein expressions were significantly decreased (P<0.05). Body weight, FSG, TG, TC, fetal body weight, serum and placental tissue MDA levels were significantly increased in the model group (P<0.05). Compared with the model group, symptoms of placental ultrastructural damage were alleviated in the low-dose formononetin group and the high-dose formononetin group, and the embryonic survival rate, serum and placental tissue GSH, SOD, TAC levels, placental tissue Nrf2, HO-1, NQO1 protein expressions were all increased (P<0.05). The body weight, FSG, TG, TC, fetal body weight, serum and placental tissue MDA levels were all decreased (P<0.05), and the effect of high dose of formononetin was stronger. Changes of all indexes in the ML385 group were contrary to those in the formononetin treatment group, and formononetin can reverse the oxidative stress damage of ML385 on GDM rats, improve the placental damage and adverse pregnancy outcome of rats. Conclusion Formononetin can reduce oxidative stress injury in GDM rats by activating Nrf2/HO-1/NQO1 pathway, thereby improving placental damage and adverse pregnancy outcomes in rats.

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Study on the effect of Shengyang Yiwei Decoction on the immune function of rats with lung cancer based on PI3K/Akt/NF-κB signal pathway

WANG Hongjun, SUN Xingxing, ZHANG Runlian

2023, 51 (7): 739-745. doi: 10.11958/20221626

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Objective To explore the effect of Shengyang Yiwei Decoction on the immune function of lung cancer rats and its possible mechanism. Methods Twelve of 72 SPF Wistar rats were randomly divided as the control group, and the remaining 60 rats were infused with carcinogenic iodized oil solution into trachea to establish lung cancer models. After modeling, model rats were divided into the model group, the cisplatin group (5 mg/kg), the Shengyang Yiwei Decoction group (14 g/kg), the phosphoinositide 3-kinase (PI3K) activator 740 Y-P group (10 mg/kg) and the Shengyang Yiwei Decoction + 740 Y-P group (Shengyang Yiwei Decoction 14 g/kg + 740 Y-P 10 mg/kg), with 12 animals in per group. After corresponding drug intervention, the proportions of peripheral blood T lymphocyte subsets CD3⁺, CD4⁺ and CD8⁺were detected by flow cytometry, and CD4⁺/CD8⁺ ratio was calculated. Enzyme-linked immunosorbent assay (ELISA) was performed to measure the serum inflammatory factors interleukin-6 (IL-6), tumor necrosis factor α (TNF-α) and immunoglobulin (IgG，IgA，IgM) levels. Samples of lung, spleen and thymus were taken and weighed to calculate the indexes of lung, spleen and thymus. HE staining was performed to observe the pathological changes of lung tissue. Real-time fluorescent quantitative PCR (RT-qPCR) was performed to measure the expression of matrix metalloproteinase-9 (MMP-9) and intercellular adhesion molecule-1 (ICAM-1) mRNA in lung tissue. Western blot assay was performed to measure PI3K/protein kinase B (Akt)/nuclear factor-κB (NF-κB) pathway related protein expression in lung tissue. Results Compared with the control group, CD3⁺, CD4⁺ proportions, CD4⁺/CD8⁺ ratio, spleen index and thymus index，IgG，IgA and IgM levels were decreased in the model group, and CD8⁺ proportion, lung index, IL-6 and TNF-α levels, lung tissue ICAM-1 mRNA and MMP-9 mRNA levels, p-PI3K/PI3K, p-Akt/Akt ratios and nuclear NF-κB p65 protein level increased (P<0.05). The left lobe of lung tissue was hyperplasia, squamous epithelial metaplasia and a large number of tumor cells were arranged disorderly in sheets or strips, accompanied by more inflammatory cell infiltration. Compared with the model group, the Shengyang Yiwei Decoction group showed increased CD3⁺ and CD4⁺ proportions, CD4⁺/CD8⁺ ratio, spleen index and thymus index, IgG，IgA，and IgM levels，and decreased CD8⁺ proportion，lung index，IL-6 and TNF-α levels, lung tissue ICAM-1 mRNA and MMP-9 mRNA levels, p-PI3K/PI3K, p-Akt/Akt ratios and nuclear NF-κB p65 protein level (P<0.05). Lung tissue lesions were reduced, lung tissue swelling was not obvious, and the structure of alveolar sacs and epithelial cells were basically clear. On the basis of 740 Y-P, Shengyang Yiwei Decoction could inhibit PI3K/Akt/NF-κB signal pathway and enhance the immunity of lung cancer rats. Conclusion Shengyang Yiwei Decoction may enhance the immune function of T lymphocytes in lung cancer rats by inhibiting PI3K/Akt/NF-κB signaling pathway.

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A clinical analysis of single row anchor combined with bone tunnel and double row anchor in the treatment of humerus greater tuberosity fracture

LIU Guoyin, LYU Dezhen, LENG Nannan, BAI Tianting, WANG Yongqiang, CHEN Jianmin, WANG Yong

2023, 51 (7): 746-750. doi: 10.11958/20221971

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Objective To investigate the clinical effect of single row anchor (SRA) combined with bone tunnel (SRA-BT) and double row anchor (DRA) in the treatment of greater tuberosity fracture (GTF) of humerus. Methods The medical records of 40 patients with GTF treated with anchor fixation technique were retrospectively analyzed. Patients were divided into the SRA-BT group (18 cases) and the DRA group (22 cases) by surgical methods after doctor-patient communication. The suture bridge technique of SRA combined with bone tunnel was used in the SRA-BT group, and DRA with suture bridge was used in the DRA group. The perioperative and postoperative indicators, complications and postoperative displacement distance of the greater tuberosity were compared between the two groups. The pain degree, functional status and range of motion were assessed 6 months after surgery. Results Bone healing was achieved in all patients without infection or internal fixation failure. There were no significant differences in operative time, intraoperative blood loss, hospital stay, postoperative complications, bone healing time and postoperative displacement distance of the greater tuberosity between the two groups (P>0.05). The incision length was larger in the SRA-BT group than that of the DRA group (P<0.05). The in-patients costs was significantly lower in the SRA-BT group than that of the DRA group (P<0.05). There were no significant differences in postoperative complications between the two groups (P>0.05). The VAS score and ASES score at rest and activity were significantly improved after operation in both groups (P<0.05), but there were no significant differences in VAS score, ASES score and ranges of motion in flexion, abduction, 0°external rotation and 90° internal rotation at rest and activity after operation between the two groups (P>0.05). Conclusion The clinical effect of technology of SRA-BT and DRA have clear clinical efficacy in the treatment of GTF, and both of them could effectively improve shoulder joint function and relieve postoperative pain. However, the SRA-BT has more advantages in reducing medical costs.

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Analysis of the urinary protein and pathological characteristics of IgA vasculitis with nephritis in children with dyslipidemia

BAI Mengke, YANG Xiaoqing, MEI Xiaofeng, LI Jin’gang, ZHANG Qiushuang, HUANG Yanjie

2023, 51 (7): 751-755. doi: 10.11958/20222046

Abstract ( 420 )

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Objective To investigate the effect of dyslipidemia on urinary protein and pathological characteristics in children with immunoglobulin vasculitis with nephritis (IgAVN). Methods A total of 316 children with IgAVN diagnosed clinically and pathologically were selected. Children with IgAVN were divided into the normal lipid group (121 cases) and the dyslipidemia group (195 cases) according to the condition of blood lipid. The dyslipidemia group was further divided into the hypercholesterolemia group (55 cases), the hypertriglyceridemia group (44 cases), the mixed hyperlipidemia group (58 cases) and the low high density lipoprotein cholesterol (HDL-C) group(38 cases). The children's sex, age, 24-hour urinary protein quantification (24 hUP), urinary protein/creatinine (UPCR), urinary immunoglobulin G/creatinine (UIgG/Cr), urinary retinol-binding protein/creatinine (URBP/Cr), urinary red blood cell count (URBC), serum albumin, total cholesterol (TC), triacylglycerol (TG) and HDL-C were collected. The renal pathology was graded according to the IgAVN grading standard of the International Study of Kidney Disease in Children. According to the Oxford pathological classification of IgA nephropathy (IgAN), children were scored semi-quantitatively. Clinical data and pathological characteristics of the normal and abnormal blood lipid groups and each subgroup were compared. Results The serum albumin level decreased in the dyslipidemia group, and the proportion of children with 24 h UP, UPCR, UIgG/Cr, URBP/Cr levels, and serum albumin<30 g/L were higher than those in the normal lipid group (P<0.05). The pathological grading of children was mainly grade Ⅲ (223 cases, 70.6%) and grade Ⅱ (82 cases, 25.6%), of which children with grade Ⅳ (11 cases, 3.8%) were all in the dyslipidemia group. The proportion of pathological grade Ⅲ+Ⅳ, diffuse mesangial hyperplasia (Mb), glomerular segmental sclerosis or adhesion (S1) was higher in the dyslipidemia group than that in the normal blood lipid group (P<0.05). Compared with the normal blood lipid group, URBP/Cr increased and serum albumin decreased in the hypercholesterolemia group, while 24 h UP, UPCR, UIgG/Cr, URBP/Cr, URBC increased and serum albumin decreased in the mixed hyperlipidemia group (P<0.05). The proportion of pathological grade Ⅲ+Ⅳ, Mb and S1 was increased in the hypercholesterolemia group and the combined hyperlipidemia group (P<0.05). Conclusion IgAVN children with hypercholesterolemia and mixed hyperlipidemia, whose renal pathological damage and proteinuria are more serious than those with normal blood lipids, should be actively treated with lipid-lowering therapy.

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The heterozygous variant c. 1574C > G (p.T525R) of SASH1 gene can promote the increase of melanin synthesis

CHEN Hongyu, ZHANG Jing, ZENG Zhen, YANG Pingping, XIONG Yu, ZHANG Miao, ZHOU Ding’an

2023, 51 (7): 756-761. doi: 10.11958/20221703

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Objective To report a case of dyschromatosis universalis hereditaria (DUH) caused by the variant site of c.1574C>G (p.T525R) of SASH1 gene and discuss the effect of mutation on melanin synthesis by functional experiments in vitro. Methods DNA was extracted from the peripheral blood of the proband and his parents, and pathogenic genes were identified by whole-exome sequencing and Sanger sequencing. Bioinformatics was used to analyze the harmful gene variant site. Mouse melanoma cell B16 was used as the cell model, and Western blot assay was used to analyze the effect of SASH1 gene c.1574C>G (p.T525R) variation on the expression of microphthalmia-associated transcription factor (MITF) and Tyrosinase proteins. In vitro melanin quantification experiment was used to analyze the effect of this variant site on melanin synthesis in human melanoma cells SK-MEL-1 and B16 cells. Results Gene sequencing results showed that the proband was detected SASH1 gene c.1574C>G (p.T525R) mutation. Bioinformatics analysis showed that the variant had a high probability affecting protein structure and function. Western blot results showed that this variant site up-regulated the expression levels of MITF and Tyrosinase, which were the most critical in melanin synthesis. This variation analyzed by in vitro melanin quantification can increase the melanin synthesis in B16 and SK-MEL-1 cells. Conclusion The heterozygous variant c.1574C>G (p.T525R) of SASH1 gene is associated with DUH, and this mutation can promote the increase of melanin synthesis in melanoma cells.

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The study on the mechanism of salidroside in the adjuvant treatment of patients with moderately severe acute pancreatitis

WANG Xiaohong, QIAN Jing, WANG Jiawei, QI Xiaoming, MENG Yun, WANG Ping, CHENG Ruizhi, ZHOU Guoxiong

2023, 51 (7): 762-765. doi: 10.11958/20221606

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Objective To observe the adjuvant therapeutic effect of salidroside on patients with moderately severe acute pancreatitis (MSAP) and explore its mechanism. Methods A total of 60 patients with MSAP were randomly divided into the routine group (30 cases) and the experimental group (30 cases) using the random number table method. A routine treatment method was given to patients in both groups, and patients in the experimental group were additionally treated with salidroside. Serum levels of cysteine-containing aspartate-specific proteases (Caspase)-1, Gasdermin D (GSDMD), absent in melanoma 2 (AIM2), interleukin (IL)-1β and IL-18 were detected by enzyme-linked immunosorbent assay (ELISA) before and 7 days after treatment in both groups. The acute physiology and chronic health evaluation (APACHE) Ⅱ and pancreas-modified CT severity index (MCTSI) scores were evaluated. The time of abdominal pain-relieving and the hospital stay were also recorded in the two groups. Results Compared with 1 d before treatment, serum levels of Caspase-1, GSDMD, AIM2, IL-1β and IL-18 decreased 7 d after treatment in both groups, and serum levels of all above indexes were significantly lower in the experimental group than those in the routine group (P<0.01). Scores of APACHE Ⅱ and pancreas-MCTSI decreased in both groups on 7 d after treatment, and scores of two indexes were significantly lower 7 d after treatment in the experimental group than those in the routine group (P<0.01). The time of abdominal pain-relieving and the hospital stay were all shorter in the experimental group than those in the routine group (P<0.01). Conclusion Salidroside can play a good adjuvant therapeutic effect on patients with MSAP by inhibiting AIM2 inflammasome-mediated pyroptosis.

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Effects of edaravone dexborneol combined with rt-PA intravenous thrombolysis therapy on efficacy and prognosis of ultra-early acute cerebral infarction

NIE Yameng, ZHANG Xiaoqiang, FENG Pengzhan, DING Yaqian

2023, 51 (7): 766-770. doi: 10.11958/20221607

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Objective To observe the efficacy and prognosis of edaravone dexborneol combined with intravenous thrombolysis with recombinant tissue plasminogen activator (rt-PA) in the treatment of ultra-early acute cerebral infarction (ACI). Methods A total of 163 patients with ultra-early ACI were randomly divided into the combination group (treated with edaravone dexborneol combined with intravenous thrombolysis with rt-PA, n=78) and the control group (treated with intravenous thrombolysis with rt-PA, n=85). Therapeutic effects and changes of superoxide dismutase (SOD), reactive oxygen species (ROS), malondialdehyde (MDA), neuron-specific enolase (NSE), matrix metalloproteinase-9 (MMP-9), matrix metalloproteinase-12 (MMP-12), central nervous system-specific protein β (S100β), low density lipoprotein cholesterol (LDL-C), total cholesterol (TC), creatine kinase (CK), creatine kinase isoenzyme (CK-MB), D-dimer (D-D), C-reactive protein (CRP), the National Institutes of Health Stroke Scale (NIHSS) scores, Modified Rankin Scale (mRS) scores and activity of daily living (ADL) scores were observed before and after treatment in the 2 groups. The fatality rate and the incidence of adverse reactions were compared between the two groups. Results After treatment, the total effective rate, SOD level and ADL score were higher in the combination group than those in the control group. Levels of ROS, MDA, NSE, MMP-9 and S100β, NIHSS score and mRS score were lower in the combination group than those in the control group (P<0.05). There were no significant differences in fatality rate and incidence of adverse reactions between the two groups (P>0.05). Conclusion Edaravone dexborneol combined with intravenous thrombolysis with rt-PA can improve the clinical effect in the treatment of patients with ultra-early ACI, and improve the prognosis.

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Short-term clinical efficacy observation of belizumab in the treatment of primary systemic lupus erythematosus

WU Chunye, XING Jun, GONG Baoqi

2023, 51 (7): 771-775. doi: 10.11958/20221006

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Objective To investigate the efficacy and safety of belimumab in the treatment of new-onset systemic lupus erythematosus (SLE). Methods Sixty patients with newly diagnosed SLE were selected and divided into 2 groups, with 30 patients in each group. One group received standard treatment, and the other group received beliumab combined with standard treatment. The indexes of the two groups before and after 10-month treatment were recorded，including white blood cell (WBC), platelet (PLT), hemoglobin (Hb), complement C3 (C3), complement C4 (C4), erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), serum white sphere ratio (AGR), immunoglobulin G (IgG), double-stranded DNA (ds-DNA), antinuclear antibody titer (ANA), 24 h urine protein quantification, prednisone acetate dosage before and after treatment and disease activity index (SLEDAI-2000). Adverse events were also recorded. Results After treatment, blood WBC, PLT, Hb, C3, C4 and AGR were higher than those before treatment in the two groups, and WBC, C3 and AGR were significantly higher in the belliumab group than those in the standard treatment group (P<0.05). IgG, ESR and ds-DNA were all decreased after treatment in the two groups of patients, and those of belliumab group were lower than the other groups after treatment (P<0.05). After treatment, doses of SLEDAI-2000 and prednisone acetate decreased in the 2 groups compared with before treatment, and those of belliumab group were lower than the standard treatment group after treatment (P<0.05). After treatment, 24 h urinary protein quantity of renal affected patients was lower in 2 groups than before treatment, and that of the belliumab group was lower than that of the standard treatment group (P<0.05). After treatment, the total effective rate was higher in the belliumab group than that of the standard treatment group (P<0.05). There was no significant difference in the incidence of adverse reactions during treatment between the two groups (P =1.000). Conclusion Beliumab can assist in reducing glucocorticoid dosage to help control disease activity without increasing adverse events, and it is recommended for the treatment of lupus patients with initial mild to moderate disease activity.

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Correlation between serum ficolin and coronary artery stenosis and its influence on prognosis in patients with coronary heart disease

LIU Shouhong, ZHANG Yongquan, WANG Qiao

2023, 51 (7): 776-780. doi: 10.11958/20221707

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Objective To investigate the correlation between Ficolin-2, Ficolin-3 and the degree of coronary artery stenosis in patients with coronary heart disease and its influence on prognosis. Methods A total of 137 patients with stable coronary heart disease were classified as the coronary heart disease group. Gensini score was used to evaluate the degree of coronary artery stenosis. According to the score, patients were divided into the mild group (56 cases), the moderate group (49 cases) and the severe group (32 cases). According to the occurrence of major adverse cardiac events (MACEs) within 3 months, patients were divided into the good prognosis group (108 cases) and the poor prognosis group (29 cases). Sixty healthy subjects in the same period were selected as the control group. The clinical data of each group were collected, and serum levels of Ficolin-2 and Ficolin-3 were detected. Pearson product moment correlation analysis was used to analyze the correlation between Ficolin-2, Ficolin-3 and Gensini. Logistic regression analysis was used to analyze the influencing factors of poor prognosis. The working characteristic (ROC) curve of subjects was used to evaluate the predictive value of Ficolin-2 and Ficolin-3 for the poor prognosis. Results The serum levels of Ficolin-2 and Ficolin-3 were lower in the coronary heart disease group than those in the control group (P<0.05). The serum levels of Ficolin-2 and Ficolin-3 in the mild group, the moderate group and the severe group were decreased successively (P<0.05). The serum Ficolin-2 and Ficolin-3 levels were negatively correlated with the Gensini score in patients with coronary heart disease (r=-0.708, -0.722, P<0.05). The number of coronary artery lesions and Gensini score were independent risk factors for poor prognosis in patients with coronary heart disease [OR=1.451 (95%CI: 1.292-1.628)]. The higher levels of Ficolin-2 and Ficolin-3 were independent protective factors [OR=0.815 (95%CI: 0.730-0.909) and 0.853 (95%CI: 0.787-0.924)]. The AUC of the combination of Ficolin-2 and Ficolin-3 in predicting the poor prognosis of patients with coronary heart disease was higher than that of the single index (Z=3.186, 2.953, P<0.05). Conclusion The serum levels of Ficolin-2 and Ficolin-3 are decreased in patients with coronary heart disease, and are negatively correlated with the degree of coronary artery stenosis. The combined detection of the two indicators can predict the poor prognosis of patients with coronary heart disease, and the sensitivity is high.

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Research progress of targeted inhibition of NLRP3 inflammasome in spinal cord injury

ZHU Jinghui, WEI Dongmin, REN Shuting, YANG Yanling, ZHAO Lin

2023, 51 (7): 781-784. doi: 10.11958/20221718

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Spinal cord injury (SCI) is a neurotraumatic disease that is difficult to cure, and common causes include falling from height, traffic accident and heavy object injury. Inflammatory response is an immune protection produced by body in response to external stimuli. Appropriate inflammatory response can protect body from endogenous and external damage. The nucleotide-binding domain-like receptor protein 3 (NLRP3) inflammasome, as the most studied inflamolosome, has significantly increased expression after SCI and mediates the inflammatory cascade. This article reviews the activation mechanism of NLRP3 and relevant measures to inhibit the activation mechanism of NLRP3 after SCI.

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