Table of Content

25 July 2023, Volume 44 Issue 7

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Dual luciferase reporter technology for verifying the targeted regulation of piRNA-DQ566704 on smad2

LEI Li, TAN Zhu-li, ZHANG Ke-li, CAO Hai-ming, LI Chang-ping

2023, 44(7):  748-753.  doi:10.3969/j.issn.1007-3205.2023.07.002

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Objective To investigate the targeted regulatory effect of piRNA-DQ566704 on the smad2 gene verified by dual luciferase reporter technology. 
Methods The 3′UTR sequence of smad2 gene was obtained from mouse hepatic stellate cell(HSC) genome, to synthesize the wild type (WT) and mutant (MUT) gene fragments of smad2 including the piRNA-DQ566704 binding site and construct the wild type vector of smad2 (pGL6-miR-smad2-3′UTR-WT+pRL-TK) and mutant (pGL6-miR-smad2-3′UTR-MUT+pRL-TK). The smad2 wild-type and mutant double luciferase reporter vector were transfected into 293T cells with piRNA-DQ566704 mimic, piRNA-DQ566704 inhibitor and negative control (NC) to detect the relative luciferase activity of each group and observe the effect of piRNA-DQ566704 on smad2 gene expression. The expression of smad2 mRNA and smad2 protein in mouse HSC transfected with piRNA-DQ566704 mimic, inhibitor and mimic NC was verified by real-time fluorescence quantitative reverse transcription polymerase chain reaction (RT-qPCR) and Western blot. 
Results The relative luciferase activity of the constructs carrying piRNA-DQ566704 mimic and smad2-3′UTR-WT was significantly decreased (P＜0.01), and there was no inhibitory effect on the relative luciferase activity of the constructs carrying smad2-3′UTR-MUT (P＞0.05). After overexpression of piRNA-DQ566704 in mouse HSC, the expression of smad2 mRNA and smad2 protein in the mimic group was significantly reduced (P＜0.001). 
Conclusion piRNA-DQ566704 can targetedly regulate the expression of smad2 gene; Therefore,the smad2 is a direct target gene of piRNA-DQ566704. 





Effects of dihydrotestosterone on spatial learning memory and synaptic plasticity in SAMP8 male mice under continuous hypoxia

HAN Shuo, LIU Xin-yuan, CUI Hui-xian, PAN Wen-sen

2023, 44(7):  754-759,791.  doi:10.3969/j.issn.1007-3205.2023.07.003

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Objective To investigate the effects of dihydrotestosterone (DHT) on spatial learning and memory and synaptic plasticity in hippocampus of SAMP8 male mice under continuous hypoxia, and to understand the effect of androgen on hippocampal synaptic plasticity in rapidly aging mice. 

Methods Forty healthy adult (5-month-old) male SAMP8 mice were randomly divided into Control group, DHT group, Hypoxia group, and DHT+Hypoxia group, with 8 mice in each group. Morris water maze was used to detect the spatial learning and memory abilities of mice. Golgi-cox staining was used to detect the density changes of synaptic dendritic spine in hippocampal CA1. RT-PCR and Western blot were used to detect the expression of related genes and proteins. 
Results The escape latency of the Hypoxia group was significantly longer than that of the Control group, while the escape latency of the DHT group was significantly shorter than that of the Control group and the Hypoxia group (P＜0.05). The target quadrant time in the DHT group was significantly longer than that in the Control group, and the number of crossing platform was significantly greater than that in the Control group (P＜0.05). The density of synaptic dendritic spine in hippocampal CA1 region in the DHT group and the DHT+Hypoxia group was significantly higher than that in Control group and the Hypoxia group (P＜0.05). Hif-1α, Epo and Epor expression of Hypoxia group was higher than that of the Control group, while the expression of Glut1, Syn, Psd-95, and Drebrin was lower than that of the Control group. The expression of Hif-1α, Erk, Glut1, Syn, Psd-95, and Drebrin genes in the DHT group was higher than that of the Control group, while the expression of Hif-1α and Epo gene of  the DHT+Hypoxia group was higher than that of the Control group (P＜0.05). Hif-1α and Epo expression in DHT group was lower than that in Hypoxia group, and Erk, Glut1, Syn, Psd-95, and Drebrin expression was higher than that in Hypoxia group; Hif-1α was lower in the DHT+Hypoxia group than that in the Hypoxia group, and Glut1, Syn, Psd-95, and Drebrin expression was higher than that in the Hypoxia group (P＜0.05). The protein expression of ERK, pERK, Glut1, and PSD95 was lower in the Hypoxia group than in the Control group, while Iba1 was higher than that in the Control group. The protein expression of ERK, pERK, Glut1, PSD95, Drebrin, SYN, and Iba1 in the DHT group was higher than that in the Control group, and the protein expression of ERK, pERK, Glut1, PSD95, and Iba1 in the DHT+Hypoxia group was higher than in the Control group (P＜0.05). The expression of ERK, pERK, Glut1, PSD95, Drebrin, and SYN proteins in the DHT group was higher than that in the Hypoxia group, while the expression of Iba1 protein was lower than that in the Hypoxia group. The expression of ERK, pERK, Glut1, and PSD95 proteins in the DHT+Hypoxia group was higher than that in the Hypoxia group (P＜0.05). 
Conclusion Dihydrotestosterone can promote the synaptic plasticity of hippocampal neurons and improve the spatial learning and memory abilities via  HIF-1α, ERK and GLUT1 signaling pathways. 








Long-chain noncoding RNA XIST regulates cell proliferation and apoptosis of mesangial cells in lupus nephritis by targeting miR-150-5p

XIAO Hui, HAN Xing, JIN Hong-jie, LIU Xiao

2023, 44(7):  760-766.  doi:10.3969/j.issn.1007-3205.2023.07.004

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Objective To observe the effects of long-chain noncoding RNA X-inactive specific transcripts (LncRNA XIST, XIST) and miR-150-5p on the proliferation and apoptosis of mesangial cells in lupus nephritis (LN) and to explore its mechanism of action. 
Methods Mesangial cells from LN mice and normal mice were isolated and cultured. pcDNA group (transfected empty vector), pcDNA-XIST group (transfected pcDNA-XIST), antagomiRNA group (transfected antagomiRNA), antagomiR-150-5p group (transfected antagomiR-150-5p), pcDNA-XIST + agomiRNA group (co-transfected pcDNA-XIST and agomiRNA), pcDNA-XIST + agomiR-150-5p group (co-transfected pcDNA-XIST and agomiR-150-5p) were transfected into mesangial cells of LN by liposome method. Real time fluorescence quantitative reverse transcription polymerase chain reaction (RT-qPCR) was used to detect the expression of XIST and miR-150-5p. Dual luciferase assay was used to detect the fluorescence activity of cells, and cell counting kit (CCK8) and annexin V-fluorescein isothiocyanate propidium iodide (annexin V-FITC/PI) were used to detect cell proliferation and apoptosis. Protein expressions of cyclin-dependent kinase inhibitor 1A (p21), proliferating cell nuclear antigen (PCNA), B-cell lymphoma-2 (Bcl-2) and BCL2-associated X protein (Bax) were detected by Western blotting (WB) . 
Results Compared with normal mouse renal tissue or mesangial cells, the expression of XIST significantly increased and the expression of miR-150-5p significantly decreased in renal tissue or mesangial cells of LN mice (P＜0.001). Overexpression of XIST or inhibition of miR-150-5p significantly facilitated the proliferation and inhibited apoptosis, up-regulated the expression of PCNA and Bcl-2 protein, and down-regulated the expression of p21 and Bax protein of mesangial cells of LN (P＜0.001). MiR-150-5p significantly inhibited the fluorescence activity of wild-type XIST cells and negatively regulated XIST (P＜0.001). Overexpression of miR-150-5p significantly inhibited the effect of overexpression of XIST on the proliferation and apoptosis of mesangial cells of LN (P＜0.001). 
Conclusion LncRNA XIST can promote the proliferation and inhibit apoptosis of mesangial cells of LN, and its mechanism may be related to directly targeting miR-150-5p. 





Mechanism of zerumbone affecting mucoepidermoid carcinoma by immunogenic cell death

SU Feng-qin, SU Hui, WU Hui-wei, XING Shu-hua

2023, 44(7):  767-772.  doi:10.3969/j.issn.1007-3205.2023.07.005

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Objective To explore the mechanism of zerumbone (Zer) in the treatment of mucoepidermoid carcinoma (MEC), in order to provide new ideas and theoretical basis for the development of MEC-related targeted drugs in clinical practice. 
Methods MEC cell lines H292 and H3118 were purchased, and the IC50 of Zer on MEC cell lines was detected by 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay. Cell proliferation was detected by colony formation assay, and cell apoptosis and surface calreticulin(CRT) protein levels were detected by flow cytometry. The expression level of highmobility group protein B1(HMGB1) was detected by Western blot, to observe whether Zer could affect the immunogenic cell death(ICD) of MEC. Finally, to explore the possible molecular regulatory mechanism of Zer,the level of signal transducer and activator of transcription 3(STAT3) in MEC cells was up-regulated by pcDNA3.1-STAT3, and the transfection efficiency was detected by qRT-PCR. The expression levels of endoplasmic reticulum stress-related factors activating transcription factor 4(ATF4), ATF3 and C/EBP-homologousprotein(CHOP) were detected by real-time quantitative PCR (qRT-PCR) and Western blot. The adenosine triphosphate (ATP) level in the medium was detected by ATP assay kit. 
Results The apoptosis rate of cells in the Zer group was higher than that in the control group (P＜0.05), and the HMGB1 protein level in the supernatant of H3118 cells in the Zer group was higher than that in the control group (P＜0.05). The ATP level in H3118 cells in the Zer group was higher than that in the control group (P＜0.05), and the protein and mRNA expression levels of endoplasmic reticulum stress-related genes ATF4, ATF3, and CHOP in the Zer group were higher than those in the control group (P＜0.05). The STAT3 mRNA level in MECs in the Zer group was lower than that in the control group (P＜0.05), and the protein and mRNA expression levels of endoplasmic reticulum stress-related genes ATF4, ATF3 and CHOP in the Zer+oe STAT3 group were lower than those in the Zer+oe NC group (P＜0.05). 
Conclusion Zer-mediated STAT3 promotes endoplasmic reticulum stress in MEC cells and induces ICD of MEC cells. 





MiR-486-5p/FOXO1 axis regulates keratinocyte proliferation and migration during repair of deep second-degree burn wound

GAO Fu-cheng, WANG Yao-jun, XUE Jia-jie, HAO Qing-ye, GAO Deng-wen, GUO Lei

2023, 44(7):  773-780,797.  doi:10.3969/j.issn.1007-3205.2023.07.006

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Objective To explore the regulatory mechanism of the miR-486-5p/forkhead box transcription factor O subfamily 1 (FOXO1) axis on the proliferation and migration of keratinocytes during repair of deep second-degree burn wound. 
Methods A mouse model of deep second-degree burn was established, and the mice were randomly divided into 4 groups: NC mimic+Vector group, FOXO1 group, miR-486-5p mimic group and miR-486-5p mimic+FOXO1 group. The wound was treated with miR-486-5p mimic and/or FOXO1 to observe the wound healing. At different time points after burn, including hemostasis (0 h), inflammation (24 h), and proliferation (7th and 14th day), in situ hybridization and reverse transcription quantitative polymerase chain reaction (RT-qPCR) were used to detect miR-486-5p expression sites and levels in wound edge tissues, respectively. The expression of FOXO1 was detected by immunofluorescence staining. Luciferase reporter analysis confirmed the binding relationship between FOXO1 and miR-486-5p. The human HaCaTkeratinocytes(HaCaT) cell model of miR-486-5p overexpression or knockdown was constructed. The proliferation and migration of cells were investigated by EdU method and scratch test, and the expression of FOXO1 protein in cells was analyzed by Western blot. 
Results miR-486-5p was significantly up-regulated during the healing of deep second-degree burn wounds in mice. In vivo model showed that miR-486-5p promoted wound healing by inhibiting the expression of FOXO1. In HaCaT cells, miR-486-5p overexpression increased proliferation and migration of the cells, where as miR-486-5p inhibition had the reverse effect. FOXO1 was a direct target of miR-486-5p in keratinocytes. FOXO1 overexpression decreased the percentage of EdU positive cells and wound healing of HaCaT cells, and reversed the induction of EdU positive cells and wound healing by miR-486-5p. 
Conclusion MiR-486-5p is a crucial regulator in repair of deep second-degree burn wound that facilitates keratinocyte proliferation and migration by inhibiting the expression of FOXO1. 





Expression and clinical significance of serum 8-iso-PGF2α, Irisin and FNDC5 in patients with vascular cognitive impairment

ZHU Qian, YAO Jie, WANG Shang-chen, CHU Li-fang

2023, 44(7):  781-785.  doi:10.3969/j.issn.1007-3205.2023.07.007

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Objective To explore the expression and clinical significance of serum 8-iso-prostaglandin (8-iso-PGF2α), Irisin, and fibronectin type Ⅲ domain-containing protein5 (FNDC5) in patients with vascular cognitive impairment (VCI). 
Methods A total of 92 patients with VCI in our hospital were selected as the observation group, and 48 healthy physical examinees were selected as the control group during the same period. The serum 8-iso-PGF2α, Irisin, FNDC5 levels and montreal cognitive assessment (MoCA) scores of the two groups were compared, and the correlation of serum 8-iso-PGF2α, Irisin, and FNDC5 levels with MoCA scores in VCI patients was analyzed. Logistic regression analysis was used to analyze the factors influencing the progression of vascular cognitive impairment no dementia (VCIND) to vascular dementia (VD). Receiver operating characteristic （ROC) curve was used to analyze the predictive value of serum 8-iso-PGF2α, Irisin, and FNDC5 in predicting VCIND progression to VD. 
Results The serum 8-iso-PGF2α level in the observation group was higher than that in the control group, while the serum Irisin, FNDC5 level and MoCA score were lower than those in the control group (t=13.999, 11.702, 20.186, 9.211, P＜0.05). The serum 8-iso-PGF2α level of VCI patients was negatively correlated with MoCA score, and the levels of Irisin and FNDC5 were positively correlated with MoCA score (r=-0.736, 0.751, 0.698, P＜0.05). Logistic regression analysis showed that elevated serum 8-iso-PGF2α and decreased Irisin and FNDC5 were independent risk factors for VCIND progression to VD (OR=4.999, 0.362, 0.387, P＜0.05). The ROC curve showed that 8-iso-PGF2α, Irisin and FNDC5 had certain predictive value for VCIND progression to VD, and the AUC of the joint prediction reached 0.866. 
Conclusion Serum 8-iso-PGF2α, Irisin and FNDC5 are all involved in the occurrence and development of VCI, which are closely related to the degree of cognitive impairment in patients, and can effectively predict the risk of progression from VCIND to VD.





Therapeutic effect of DEVT and BT on patients with acute large vessel occlusive cerebral infarction and their impact on prognosis

WANG Lin, XU Zheng-hu, LI Ke-jing, FAN Ming-yue, WANG Heng

2023, 44(7):  786-791.  doi:10.3969/j.issn.1007-3205.2023.07.008

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Objective To investigate the therapeutic effects of direct endovascular therapy (DEVT) and bridging therapy (BT) on patients with acute large vessel occlusion cerebral infarction and their impact on prognosis. 
Methods A prospective randomized study was conducted to select 120 patients with acute large vessel occlusion cerebral infarction and divide them into DEVT group (n=60) and BT group (n=60) according to random number table method. DEVT group was given DEVT, and BT group was given BT. The scores of the National Institutes of Health Stroke Scale (NIHSS), Barthel index (BI), and modified Rankin score (mRs) were observed before and after treatment in both groups. In the meantime, serum brain derived neurotrophic factor (BDNF), nerve specific enolase (NSE), insulin-like growth factor-1 (IGF-1), high-sensitivity C-reactive protein (hs-CRP), tumor necrosis factor-α (TNF-α), endothelin-1 (ET-1), nitric oxide (NO) and calcitonin gene-related peptide (CGRP) levels were detected in the two groups. 
Results There was no significant difference in the proportion of patients with mRS score ≤ 2 between two groups (P＞0.05). At one week after treatment, the NIHSS score and NSE in the two groups were significantly lower than those before treatment, while the levels of BDNF and IGF-1 were higher than those before treatment. The NIHSS score and NSE in the BT group were significantly lower than those in the DEVT group, while the BDNF and IGF-1 were significantly higher than those in the DEVT group (P＜0.05). At 24 h after treatment, the levels of hs-CRP, TNF-α, and ET-1 in the two groups were lower than those before treatment, while the levels of NO and CGRP were higher than those before treatment. The levels of ET-1 in the BT group were lower than those in the DEVT group, while the levels of NO and CGRP were higher than those in the DEVT group (P＜0.05). 
Conclusion Both DEVT and BT regimens have significant effects in the treatment of patients with acute large vessel occlusion cerebral infarction. BT regimens are more conducive to the recovery of neurological function and improvement of vascular endothelial function.  





Effect of tirofiban combined with Solumbra embolization on the vascular recanalization rate and prognosis of acute cerebral infarction caused by intracranial and extracranial large artery occlusion

WANG Rong-hui, WANG Xiao-wei, ZHANG Bo-gang

2023, 44(7):  792-797.  doi:10.3969/j.issn.1007-3205.2023.07.009

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Objective To investigate the application of tirofiban combined with Solumbra technique for mechanical embolization in acute cerebral infarction caused by intracranial and extracranial large artery occlusion, and to analyze its effects on vascular recanalization rate, platelet function and prognosis. 
Methods A total of 120 patients with acute cerebral infarction caused by intracranial and extracranial large artery occlusion were selected, and they were grouped according to different medications used after intervention, with 60 patients in each group. Both groups underwent mechanical embolization by Solumbra technique, the control group was given aspirin combined with clopidogrel after embolization, and the observation group was given tirofiban after embolization. The clinical efficacy, vascular recanalization rate, cerebral blood flow ［meanblood flow velocity (Vm) of middle cerebral artery, peak systolic flow velocity (Vs), end-diastolic flow velocity (Vd)］, platelet function indexes ［mean platelet volume (MPV), platelet aggregation rate (PAgT), platelet adhesion rate (PAdT), platelet P-selectin］, neurological function ［National Institute of Health Stroke Scale (NIHSS)］ score, activities of daily living ［Barthel index (BI) score］, prognosis ［modified rankin scale (mRs) score］ and incidence of bleeding complications, and mortality were compared between two groups. 
Results The total effective rate and vascular recanalization rate of the observation group were higher than those of the control group (P＜0.05). The Vm, Vs, and Vd of the middle cerebral arteries in both groups increased with the duration of treatment, and the observation group showed a more significant increase; The difference between groups, time points, and time points between groups was statistically significant (P＜0.05). The levels of MPV, PAgT, PAdT, and P-selectin in the two groups decreased with the prolongation of treatment, and the observation group showed a more significant decrease. The difference between groups, time points, and time points between groups was statistically significant (P＜0.05). The NIHSS score and mRS score of the two groups decreased with the prolongation of treatment, and the observation group showed a more significant decrease. The Barthel index increased with the duration of treatment, and the observation group showed a more significant increase; The difference between groups, time points, and time points between groups was statistically significant (P＜0.05). There was no statistically significant difference in the incidence of bleeding complications and mortality rate between two groups at 90 d after treatment (P＞0.05). 
Conclusion The application of tirofiban combined with Solumbra technique for mechanical embolization in patients with acute cerebral infarction caused by intracranial and extracranial large artery occlusion is effective in improving platelet function and cerebral blood flow, increasing vascular recanalization rate, and promoting the recovery of neurological function, activities of daily living and prognosis, without increasing rate of bleeding complications and mortality rate.





Correlation and significance of brainstem auditory evoked potentials and cerebral blood flow in patients with epilepsy secondary to cerebral infarction

HE Qin, YANG Xiao-fang, KANG Yu, YIN Cheng-kun

2023, 44(7):  798-802,823.  doi:10.3969/j.issn.1007-3205.2023.07.010

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Objective To investigate the correlation and significance of brainstem auditory evoked potentials (BAEP) and cerebral blood flow (CBF) in patients with epilepsy secondary to cerebral infarction. 
Methods A total of 80 patients with epilepsy secondary to cerebral infarction were selected as the research group, and another 80 patients without epilepsy secondary to cerebral infarction during the same period were selected as the control group. The general information, CBF, serum lipoprotein-associated phospholipase A2 (Lp-PLA 2), neuron-specific enolase (NSE) levels and BAEP scores at admission were compared between two groups. The BAEP scores of patients with different epileptic characteristics (seizure type, discharge type, discharge site) in the research group were compared, and the correlation between BAEP scores and CBF, serum Lp-PLA2, and NSE levels in patients with epilepsy secondary to cerebral infarction was analyzed, and BAEP scores in patients with different characteristics of epilepsy secondary to cerebral infarction were compared. The correlation between BAEP score and the characteristics of epilepsy secondary to cerebral infarction was analyzed, and the value of BAEP scores, CBF, Lp-PLA2, and NSE in predicting epilepsy secondary to cerebral infarction was analyzed by receiver operating characteristic (ROC) curve. 
Results CBF was lower in the research group than in the control group, while serum Lp-PLA2 and NSE levels and BAEP scores were higher than those in the control group (P＜0.05). BAEP scores were negatively correlated with CBF and positively correlated with serum Lp-PLA2 and NSE levels in the research group (P＜0.05). BAEP scores were higher in patients with generalized seizures than in patients with partial seizures in the research group, and BAEP scores were higher in patients with multifocal discharge than in focal discharge patients (P＜0.05). BAEP scores were positively correlated with seizure type and discharge type in patients with epilepsy secondary to cerebral infarction (P＜0.05). The area under the curve (AUC) of BAEP scores, CBF, Lp-PLA2, and NSE in predicting epilepsy secondary to cerebral infarction was > 0.7, in which BAEP scores combined with CBF, Lp PLA2, and NSE in prediction had the largest AUC of 0.928 (95%CI: 0.876-0.963). 
Conclusion BAEP in patients with epilepsy secondary to cerebral infarction is closely correlated with CBF and serum Lp-PLA2 and NSE levels, and the combination of the indicators has high predictive efficacy in predicting epilepsy secondary to cerebral infarction.





Clinical effect of bedside ultrasound-guided fluid resuscitation on patients with sepsis in ICU

YI Shao-long, HUANG Hu, ZHANG Yan-lin

2023, 44(7):  803-808.  doi:10.3969/j.issn.1007-3205.2023.07.011

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Objective To analyze the clinical effect of bedside ultrasound-guided fluid resuscitation on patients with sepsis in intensive care unit (ICU). 
Methods A total of 97 sepsis patients in ICU of our hospital were selected and randomly divided into observation group (n=49) and control group (n=48) by a random number generation method. Both groups were treated with liquid resuscitation therapy. The control group was given conventional monitoring methods to guide liquid resuscitation, while the observation group was given bedside ultrasound to guide liquid resuscitation based on the control group. The volume of fluid resuscitation was compared between two groups at 6 h and 24 h after treatment. Central venous oxygen saturation (ScvO2), central venous pressure (CVP), mean arterial pressure (MAP), tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), blood lactate concentration, base residue (BE), oxygen consumption (VO2), and oxygen supply (DO2) levels were compared between two groups before treatment, and at 6 h and 24 h after treatment. The incidence of pulmonary edema, rate of requiring renal replacement therapy, mortality within one month, length of ICU stay, and length of hospital stay were statistically analyzed in both groups. 
Results At 6 h and 24 h after treatment, the amount of fluid resuscitation in the observation group was significantly less than that in the control group, suggesting significant difference (P＜0.05). With the prolongation of treatment time, the levels of ScvO2, CVP, MAP, BE, VO2, and DO2 in both groups showed a gradual upward trend, and the levels of ScvO2, CVP, MAP, BE, VO2, and DO2 in the observation group were higher than those in the control group, with statistically significant differences between groups and time points (P＜0.05). With the prolongation of treatment time, the levels of tumor necrosis factor-α, IL-6 and blood lactic acid gradually decreased, which were lower in the observation group than in the control group, and the differences between groups and time points were statistically significant (P＜0.05). The observation group had shorter ICU stay and hospital stay than the control group, and the rate of requiring renal replacement therapy and the incidence of pulmonary edema were lower than those in the control group, with a statistically significant difference (P＜0.05). There was no significant difference in mortality within one month between two groups (P＞0.05). 
Conclusion Bedside ultrasound-guided fluid resuscitation in patients with sepsis can reduce fluid rehydration volume, inhibit inflammatory reaction, improve hemorheology and oxygen metabolism indicators, and shorten the length of ICU stay, and it plays a positive role in maintaining acid-base balance and promoting patients′ rehabilitation. 





Clinical value of dual-channel negative pressure aspirator used in laparoscopic radical resection of rectal cancer

DING Hong-rui, LIU Teng-fei, YANG Liu, CAO Yue-peng, LI Peng-yi, QIAN Chen

2023, 44(7):  809-813.  doi:10.3969/j.issn.1007-3205.2023.07.012

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Objective To investigate the clinical value of a new dual-channel negative pressure aspirator in laparoscopic radical resection of rectal cancer. 
Methods A total of 102 patients who initially underwent laparoscopic radical resection of rectal cancer in our hospital were randomly divided into observation group (n=51)and control group (n=51) according to random number table. Patients in the two groups were operated by the same operation team. The conventional single-channel negative pressure aspirators were used in the laparoscopic resection of patients in control group, and new dual-channel negative pressure aspirators were used in the laparoscopic resection of patients in observation group. Continuous negative pressure suction of surgical smoke and excess CO2 in the body of patients during surgery was performed. The surgical smoke removal effect, operation condition, postoperative recovery and complication incidence rate were compared between two groups. 
Results The times of lens wiping in observation group were significantly decreased than those in control group (P＜0.05), and the quality of visual field in observation group was significantly superior to that in control group (P＜0.05). The digestive tract reconstruction time, duration of operation, intraoperative bleeding and ETCO2 after pneumoperitoneum creation in observation group were significantly lower than those in control group (P＜0.05). The postoperative drainage in observation group was significantly lower than that in control group (P＜0.05), and the first time of anal exhaust, the first time of off-bed activity, the duration of indwelling catheter and the length of postoperative hospitalization had the decreasing tendency, as compared with those in control group, showing no significant difference (P＞0.05). No significant difference was found in the total incidence of postoperative complications in control group and observation group (P＞0.05). 
Conclusion The application of dual-channel negative pressure aspirator in laparoscopic radical resection of rectal cancer has a good effect of eliminating surgical smoke, reducing the times of lens wiping and improving the quality of visual field, which is helpful to shorten the operation process without increasing postoperative complications incidence, and has a high clinical value.





Expression of STING in bladder cancer tissues and its mechanism of regulating phosphorylation modification of IFN- γ/STAT1 signaling pathway

WANG Yong, WANG Zhi-rong, LU Hai-bing, LI Mao-lin, DING Yan

2023, 44(7):  814-818,829.  doi:10.3969/j.issn.1007-3205.2023.07.013

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Objective To investigate the expression of stimulator of interferon genes (STING) in patients with bladder cancer and its mechanism of regulating interferon- γ (IFN-γ)/signal transducer and activator of transcription 1 (STAT1) signaling pathway. 
Methods In this cohort study, 110 patients with bladder cancer were selected to collect cancer tissues and adjacent tissues, and the expression of STING in cancer tissues and adjacent tissues was detected by quantitative real-time polymerase chain reaction (qPCR). In cytological test, CRISPR-Cas9 technology was used to knock out the STING gene of human bladder cancer BIU-87 cells, and bladder cancer cells were divided into three groups: control group, NC group (transfected with NC plasmid) and experimental group (transfected with CRISPR-Cas9 plasmid). MTT method was used to detect the proliferation ability of cells at different time points (at 24 h, 48 h and 72 h), and flow cytometry was used to detect the proportion of cell survival, early apoptosis and late apoptosis in each group. qPCR was used to detect mRNA expression levels of IFN- γ and STAT1, and Western blot method for detecting IFN- γ and STAT1 protein expression level as well as the phosphorylation modification level of STAT1 protein. 
Results In cohort study, the expression level of STING in bladder cancer tissue was higher than that in adjacent tissue (P＜0.05). Cytological test showed that, compared with the control group, the proliferation ability of cells in the knockout STING group decreased, and further decreased with time (P＜0.05). Cell proliferation decreased cell survival, and the proportion of early and late apoptosis increased (P＜0.05). IFN- γ and STAT1 mRNA expression levels decreased (P＜0.05), and the expression level of IFN- γ and STAT1 protein as well as the phosphorylation level of STAT1 decreased (P＜0.05). 
Conclusion STING may activate the protein expression of IFN-γ/STAT1 signaling pathway and phosphorylation modification of STAT1, thereby getting involved in the occurrence and development of bladder cancer. 





Correlations between the coronal plane parameters of trunk and Cobb angle in adolescent idiopathic scoliosis

GUO Xiao-qi, ZHU Fei-long, ZHANG Cun-gang, ZHANG Ming, SUI Yi-chen, ZHU Qian

2023, 44(7):  819-823.  doi:10.3969/j.issn.1007-3205.2023.07.014

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Objective To explore the relationship between the coronal plane parameters of trunk and Cobb angle of adolescent idiopathic scoliosis (AIS), for the purpose of providing assistance for early screening and monitoring of conservative treatment of AIS. 
Methods Forty-three AIS patients with unilateral right thoracic or left lumbar “C”-shaped curvature admitted to Department of Rehabilitation Engineering of Xuzhou Rehabilitation Hospital were selected, including 20 patients with right thoracic curvature and 23 patients with left lumbar curvature. The Cobb angle on the full length of spine X-ray, the apex vertebral translocation (AVT), trunk shift (TS), trunk shift ratio (TSR) and apex trunk rotation (ATR) degree of vertebral body in physical appearance were measured. The correlations between the AVT, TS, TSR, ATR and Cobb angle were analyzed respectively. 
Results The AVT and TSR were significantly correlated with Cobb angle, the AVT was better correlated with Cobb angle (r=0.749, P＜0.05), and the correlation between AVT, TSR and Cobb angle was greater in patients with left lumbar curvature than in patients with right thoracic curvature. There was no correlation between ATR and Cobb angle in patients with left lumbar curvature (P＞0.05). 
Conclusion There is a certain correlation between the coronal plane parameters of trunk and the Cobb angle in patients with unilateral right thoracic or left lumbar “C”-shaped curvature in AIS patients. 





Mechanisms of miRNA promoting invasion and metastasis in ovarian cancer

WANG Lu, SHAN Jia-jia

2023, 44(7):  824-829.  doi:10.3969/j.issn.1007-3205.2023.07.015

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Objective To investigate the mechanism of miRNA promoting invasion and metastasis of ovarian cancer. 
Methods By analyzing the miRNA expression profile of ovarian cancer in GSE119055 dataset of high-throughout gene expression omnibus (GEO), we screened the differential miRNAs. Prognostic miRNAs and prognostic differential genes were screened using the cancer genome atlas program (TCGA) database. The effect of heterogeneous expression of prognostic miRNA on the migration and invasion of ovarian cancer cells was detected by wound healing test and Transwell method. Bioinformatics website Starbase was used to predict the likelihood of targeted binding of prognosis related miRNAs and prognostic related genes. 
Results There were 32 differential miRNAs in the GSE119055 dataset, 20 up-regulated miRNAs and 12 down-regulated miRNAs, among which the prognostic related miRNAs were miR-187-3p and miR-381-5p. The results of wound healing test and Transwell showed that overexpression of miR-187-3p or inhibition of miR-381-5p could significantly inhibit the migration and invasion of ovarian cancer cells. Starbase predicted that there were 29 potential target genes of miR-187-3p and miR-381-5p. 
Conclusion miR-187-3p and miR-381-5p are associated with the prognosis of ovarian cancer. miR-187-3p and miR-381-5p provide a new theoretical basis for the treatment of ovarian cancer as potential target genes promoting invasion and metastasis.





A study of delineation of pelvic lymph node drainage area in intensity-modulated radiotherapy for gynecologic malignancies

HUANG Rong, LI Dan, PANG Hao-wen, DU Xiu-ju, CHEN Xiao, WEN Qing-lian

2023, 44(7):  830-835.  doi:10.3969/j.issn.1007-3205.2023.07.016

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Objective To evaluate changes in small intestine position during whole pelvic intensity-modulated radiation therapy for patients with gynecologic malignancies using computed tomographic (CT) images during treatment and to investigate whether the small intestine should be avoided for delineation of pelvic lymph node drainage area in patients with gynecologic malignancies. 
Methods Thirty-one patients with gynecologic malignancies were included for pelvic radiotherapy and divided into surgical group (n=15) and non-surgical group (n=16). The 10th, 15th, and 20th fractions of the CT images during treatment were reviewed and changes in the avoidance area formed by the intersection of the observation area with the small intestine compared with the initially planned CT scan images were recorded. We defined the scope of CTV radiation therapy in the drainage area of the pelvic lymph nodes formed by pelvic vascular expansion in the initially planned scan images as the observation area and its intersecting volume with the small bowel as avoidance area of the small intestine. 
Results During radiotherapy, the volume ratio of small intestine staying in its avoidance area of planned target area in the observation area, which included the left common iliac region, right common iliac region, left internal iliac region, right internal iliac region, left external iliac region, right external iliac region, left obturator region, right obturator region, and presacral region, was 16.52%, 28.05%, 18.65%, 37.16%, 39.18%, 49.67%, 23.74%, 50.07%, and 42.65%, respectively, for the non-surgical group, and 39.57%, 34.41%, 18.45%, 31.21%, 44.21%, 43.39%, 31.32%, 49.67%, and 27.59%, respectively, for the surgical group. 
Conclusion The volume ratio of small intestine staying in its avoidance area of planned target area accounted for 50% or less in patients during radiotherapy, regardless of whether they had received surgery for gynecological malignancies. Combined with the mobility of the intestine itself and the percentage of overlapping volume, it is recommended that the small intestines do not need to be avoided for delineation of drainage area of pelvic lymph nodes for gynecological malignancies.





Relationship between Mir-93-5P expression and Th1/Th2 balance in peripheral blood of children with RSV lower respiratory tract infection

ZHOU Lian-hua, LIU Lin, YU Qing-le, WU Lei

2023, 44(7):  836-839,850.  doi:10.3969/j.issn.1007-3205.2023.07.017

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Objective To study the relationship between the expression of microrNA-93-5P and T-helper 1 (Th1)/T-helper 2 (Th2) balance in peripheral blood of children with respiratory syncytial virus (RSV) lower respiratory tract infection.  
Methods A total of 58 children with RSV lower respiratory tract infection who were registered and treated in the Department of Pediatrics of our hospital were selected as the RSV group, and 60 age- and gender-matched healthy children were selected as the control group. The serum miR-93-5p level was detected by real-time quantitative polymerase chain reaction (RT-qPCR). The percentage of Th1 and Th2 were counted by flow cytometry, and the expression of Th1/Th2 was calculated. The levels of interleukin (IL)-2, IL-4, IL-10 and interferon-γ (IFN-γ) were detected by enzyme linked immunosorbent assay (ELISA), and the relationship between miR-93-5p and Th1/Th2 was analyzed by Spearman. Logistics regression analysis was performed to analyze the risk factors of RSV lower respiratory tract infection. 
Results The level of miR-93-5p in RSV group was higher than that in control group (P＜0.05). Th1 and Th1/Th2 in RSV group were lower than those in control group (P＜0.05), while Th2 in RSV group was higher than that in control group; the levels of IL-2 and IFN-γ were lower in RSV group than in the control group (P＜0.05). The levels of IL-4 and IL-10 in RSV group were higher than those in control group (P＜0.05). miR-93-5p inRSV group was negatively correlated with IL-2, IFN-γ and Th1/Th2 ratio (P＜0.05), and positively correlated with IL-4 and IL-10 (P＜0.05). Logistic regression analysis showed that high level of miR-93-5p and low level of Th1/Th2 were risk factors for RSV lower respiratory tract infection (P＜0.05). 
Conclusion Mir-93-5P is highly expressed in peripheral blood of children with RSV lower respiratory tract infection, and its high expression may be closely related to Th1/Th2 balance. 





Expression and clinical significance of blood lactic acid, CCSP and IL-22R in children with pneumonia

WEN Bo, LI Ling, FENG Yang, LUO Li

2023, 44(7):  840-843,865.  doi:10.3969/j.issn.1007-3205.2023.07.018

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Objective To investigate the expression and clinical significance of blood lactic acid, Clara cell secretory protein (CCSP) and interleukin-22 receptor (IL-22R) in children with pneumonia. 
Methods A total of 150 children with pneumonia treated in our hospital were selected as the observation group. They were divided into 96 cases in the acute phase and 54 cases in the convalescent phase according to the course of disease, and were divided into mild disease group and severe disease group according to the severity of the disease. Another 80 healthy children during the same period were selected as the control group. The levels of blood lactic acid, CCSP, and IL-22R in the control group, acute phase and convalescent phase were compared, and these levels in children with different conditions in the acute phase and convalescent phase were compared. The correlation between the levels of blood lactic acid, CCSP and IL-22R in children with pneumonia and the value of blood lactic acid, CCSP and IL-22R alone and in combination in the diagnosis of pneumonia were analyzed, and the receiver operating characteristic (ROC) curve was drawn. 
Results Blood lactic acid and IL-22R in the control group were lower than those in the acute and convalescent phase groups, and lower in the convalescent group than in the acute group (P＜0.05). CCSP in the control group was higher than that in the acute and convalescent groups, and higher in the convalescent group than in the acute group (P＜0.05). In the acute phase group, the levels of blood lactic acid and IL-22R in severe group A were higher than those in mild group A, while CCSP was lower than that in mild group A (P＜0.05). In the convalescent group, the levels of blood lactic acid and IL-22R in severe group B were higher than those in mild group B, while CCSP was lower than that in mild group B (P＜0.05). Pearson correlation analysis showed that blood lactic acid and IL-22R were negatively correlated with CCSP in children with acute pneumonia (r=-0.75, 0.81, P＜0.05), and blood lactic acid was positively correlated with IL-22R (r=0.06, P＜0.05). There was no correlation between blood lactic acid, CCSP and IL-22R in children in convalescent phase (P＞0.05). The area under the ROC curve (AUC) of blood lactic acid, CCSP and IL-22R in combination in the diagnosis of pediatric pneumonia was higher than that of single detection (P＜0.05). 
Conclusion The expression of blood lactic acid and IL-22R is high, while the expression of CCSP is low in children with pneumonia. The abnormal levels can indicate severe pneumonia. The combined detection of blood lactic acid, CCSP and IL-22R has high value in the diagnosis of children with pneumonia.





Value of integrated carotid ultrasonography and transcranial Doppler in perioperative period of carotid endarterectomy

LI Jun-qing, LI Dan, CHAI Jian-shuang, WANG Yan, SONG Zhi-jun, LI Hai-fen

2023, 44(7):  844-850.  doi:10.3969/j.issn.1007-3205.2023.07.019

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Objective To investigate the value of integrated carotid ultrasonography and transcranial Doppler (TCD) in perioperative period of carotid endarterectomy (CEA). 
Methods The clinical data of 166 patients who received CEA due to carotid stenosis were retrospectively analyzed. In addition to digital subtraction angiography (DSA) and/or computed tomography angiography (CTA), carotid artery color Doppler ultrasound (referred to as carotid artery ultrasound) was also conducted to evaluate carotid artery stenosis and characteristics of plaques. Transcranial color-coded Duplex (TCCD) and TCD were carried out to determine the compensatory status of collateral circulation of the brain. TCD was used to monitor the peak systolic velocity (PSV) of bilateral middle cerebral artery (MCA) during the operation, and the blood pressure was regulated individually according to the changes of PSV, so as to decide whether to use CEA shunts to prevent the occurrence of cerebral hyperperfusion syndrome. Integrated carotid ultrasonography and TCD were used for reexamination and follow-up to evaluate the effect of CEA after the operation. 
Results Carotid artery stenosis and collateral circulation in all patients were evaluated through integrated carotid ultrasonography and TCD. Among them, there were 8 cases of subtotal occlusion of the internal carotid artery, 15 cases of stenosis rate ranging from 50% to 69% with clinical symptoms, 136 cases of stenosis rate ranging from 70% to 99% (including 4 cases of stent restenosis with stenosis rate ranging from 70% to 99%), and 6 cases of carotid web. Anterior communicating artery patency was present in 27 cases, posterior communicating artery patency in 38 cases, anterior-posterior communicating artery patency in 13 cases, and internal-external carotid artery collateral circulation was open in 42 cases, and no collateral artery opening in 9 cases, which was highly consistent with DSA or CTA results. Twenty-four cases underwent CEA with shunting. Among the patients who underwent CEA without a shunt, 6 cases of mild cerebral hyperperfusion syndrome and 1 case of mild cerebral infarction occurred after operation. Within 7 days after operation, 2 cases died, including 1 death from myocardial infarction and 1 death from sudden cardiac arrest. Internal carotid artery (ICA) occlusion occurred in one case who underwent ligation of ICA due to severe carotid artery stenosis with extracranial ICA aneurysm. ICA patency was found in all the other 163 patients in the first 6-month follow-up after operation. 
Conclusion The degree of carotid artery stenosis and the opening of collateral circulation assessed by integrated carotid ultrasonography and TCD were highly consistent with DSA. Integrated carotid ultrasonography and TCD can provide reliable information before, during and after CEA, improve the safety of surgery, and has important application value in the perioperative period of CEA. 





The diagnostic efficacy of ultrasound scoring method, combined with human chorionic gonadotropin β, alpha-fetoprotein, and creatine kinase on dangerous placenta previa combined with placenta implantation

JI Li, ZHANG Xue-fang, ZHAI Xing-rong

2023, 44(7):  851-855.  doi:10.3969/j.issn.1007-3205.2023.07.020

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Objective To explore the diagnostic efficacy of ultrasound scoring method combined with human chorionic gonadotropin β (β-hCG), alpha fetoprotein (AFP), and creatine kinase (CK) on pernicious placenta previa (PPP) combined with placental implantation. 
Methods A total of 130 PPP patients were enrolled as the observation group, and 128 healthy parturients as control group. The observation group was divided into two groups according to presence of placental implantation, namely, implantation group (with placental implantation, n=97) and non-implantation group (without placental implantation, n=33). The ultrasound scoring method and the serum β-hCG, AFP, and CK levels were compared in each group. Spearman analysis was used to analyze the correlation of ultrasound scoring method and serum β-hCG, AFP, and CK with PPP and placental implantation. Receiver operating characteristic (ROC) curve was used to analyze the diagnostic efficacy of ultrasound scoring method, β-hCG, AFP, and PK levels for PPP with placental implantation. 
Results The ultrasound score method, β-hCG, AFP, and CK levels were higher in the observation group than in the control group (P＜0.05). The ultrasound score method, β-hCG, AFP, and CK in the implantation group were all higher than those in the non-implantion group (P＜0.05). The results of Spearman analysis showed that ultrasound scoring, β-hCG, AFP, and CK were positively correlated with PPP and placental implantation (r=0.566,0.537,0.439 and 0.524; P＜0.05). The results of ROC curve analysis showed that the area under the ROC curve (AUC) of ultrasound scoring method, β-hCG, AFP, CK and the combined diagnoses of the four indicators were 0.816, 0.877, 0.866, 0.758 and 0.956, respectively, and the diagnostic efficacy of the combined diagnosis of the four indicators for PPP with placental implantation was better than the single detection. 
Conclusion The combination of ultrasound scoring method with serum β-hCG, AFP, and CK has high diagnostic efficacy for PPP with placental implantation.