Tianjin Medical Journal

Study on the mechanism of lnc-NONHSAT074080 regulating target gene ARL6IP4 involved in the pathogenesis of RA-UIP

LIU Yuan, LU Fuai, WANG Le, YANG Youguo, LI Xiaofen, XIAO Yunping

2023, 51 (1): 1-7. doi: 10.11958/20220644

Abstract ( 610 )

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Objective To investigate the role and regulatory mechanism of lnc-NONHSAT074080 on the pathogenesis of rheumatoid arthritis (RA)-related common interstitial pneumonia (UIP). Methods According to whether combined with UIP, patients were divided into the RA pulmonary fibrosis group (n = 4) and the RA control group (n = 4). The long non coding RNA (lncRNA) related RA pulmonary fibrosis was analyzed by Affymetrix gene chip. The bioinformatics analysis was carried out, and lncRNAs with significantly up-regulated expression were verified by real-time auantitative polymerase chain reaction (qPCR). The lncRNA, which was verified by qPCR and showed significantly increased expression in RA pulmonary fibrosis, bioinformatics analysis showed the relationship with the process of pulmonary fibrosis, was selected as the target lncRNA. Then target lncRNA shRNA was constructed，as well as the relationship between the target lncRNA and the target gene was verified by luciferase gene report. Moreover, TGF-β1-induced human embryonic lung fibroblasts were transfected with target lncRNA shRNA. CCK8 was used to detect cell proliferation, and immunofluorescence was used to detect the expression of α smooth muscle actin (α-SMA). qPCR was used to detect the expression of target lncRNA and target genes, and enzyme-linked immunosorbent assay was used to detect the expression of ? collagen and fibronectin in cells. Results There were 192 significant upregulated lncRNA in the whole blood of RA pulmonary fibrosis group compared to the RA control group. qPCR showed that the expression of lnc-NONHSAT074080 in RA pulmonary fibrosis group was significantly increased (P<0.05). Bioinformatics analysis and luciferase gene report showed that lnc-NONHSAT074080 was related to fibrosis progression. The possible target gene including ARL6IP4 may play a role through fibrosis related pathway such as mitogen-activated protein kinase (MAPK) and signal transducer and activator of transcription (STAT) signal pathways. Compared with the blank control group, the proliferation level, α-SMA, lnc-NONHSAT074080, ARL6IP4, type I collagen and fibronectin expression levels were significantly increased in the TGF-β1 group (P<0.05). Compared with the TGF-β1 group, the TGF-β1+NONHSAT074080 shRNA group showed the opposite trend (P<0.05). Conclusion lnc-NONHSAT074080 is associated with the pathogenesis and fibrosis process of RA-UIP, and may participate in fibrosis in the fibrosis process by regulating the target gene ARL6IP4.

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Effects of T-cadherin silencing on ox-LDL induced biological behavior of human chorionic trophoblast cells HTR-8/SVneo

WANG Haijiao, HE Hongmei, QI Lin, CUI Yujiao, XIAO Chunhui, WANG Yi

2023, 51 (1): 8-13. doi: 10.11958/20220832

Abstract ( 462 )

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Objective To investigate the effect of silencing T-cadherin on oxidized low density lipoprotein (ox-LDL) induced biological behavior of human chorionic trophoblast HTR-8/SVneo cells. Methods Human chorionic trophoblast cells HTR-8/SVneo were divided into the blank control group, the ox-LDL group, the T-cadherin small interfering RNA (siRNA) negative control group and the T-cadherin siRNA group. After each group was treated accordingly, the expression levels of T-cadherin messenger RNA (mRNA) in HTR-8/SVneo cells were detected by real-time fluorescence quantitative PCR in the four groups. The proliferation of cells in each group was detected by tetramethylazolate method. The clone formation of each group was detected by plate clone formation experiment. The apoptosis of each group was detected by flow cytometry. Transwell chamber test and scratch test were used to detect the invasion and migration ability of cells. The protein expression levels of Caspase-3, Bcl-2 related X protein (Bax), B cell lymphoma-2 (Bcl-2) and matrix metalloproteinase-2 (MMP-2), matrix metalloproteinase-9 (MMP-9) of cells in each group were detected by Western blot assay. Results Compared with the blank control group, expression levels of T-cadherin mRNA, proliferation inhibition rate, apoptosis rate, Caspase-3 and Bax protein were increased significantly in the ox-LDL group and the T-cadherin siRNA negative control group, and expression levels of clone formation rate, number of invasive cells, scratch healing rate, Bcl-2, MMP-2 and MMP-9 protein were decreased significantly (P<0.05). Compared with the ox-LDL group and the T-cadherin siRNA negative control group, expression levels of T-cadherin mRNA, proliferation inhibition rate, apoptosis rate, Caspase-3 and Bax protein were decreased significantly in the T-cadherin siRNA group, and expression levels of clone formation rate, number of invasive cells, scratch healing rate, Bcl-2, MMP-2 and MMP-9 protein were increased significantly (P<0.05). Conclusion Silencing T-cadherin can inhibit ox-LDL induced abnormal apoptosis of HTR-8/SVneo cells, and promote cell proliferation, invasion and migration.

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The mechanism of miR-891a-5p regulating the biological process of colorectal cancer cells by targeting CPEB1

ZHANG Xinyan, ZHAO Guodong

2023, 51 (1): 14-18. doi: 10.11958/20220657

Abstract ( 453 )

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Objective To explore the mechanism of miR-891a-5p regulating the proliferation, apoptosis, migration and invasion of colon cancer cells through cytoplasmic polyadenylation element binding protein 1 (CPEB1). Methods Real-time fluorescence quantitative reverse transcription polymerase chain reaction (qPCR) was used to detect the expression of miR-891a-5p in colon cancer tissue, paracancer tissue, human normal colon epithelial cells and human colon cancer cells. LOVO cells were divided into the group A (without any treatment), the group B (transfected with anti-miR-con), the group C (transfected with anti-miR-891a-5p), the group D (co-transfected with anti-miR-891a-5p and si-con), group E (co-transfected with anti-miR-891a-5p and si-CPEB1), the group F (transfected with miR-con) and group G (transfected with miR-891a-5p). Cell proliferation rate was detected by CCK-8 assay, and apoptosis rate was detected by Annexin V-FITC/PI assay. Cyclin D1, Pro-caspase-3, C-caspase-3, N-cadherin, E-cadherin and Vimentin were detected by Western blot assay. Transwell assay was used to detect cell migration and invasion. Double luciferase reporter gene assay was used to detect the fluorescence activity of cells. Results The expression of miR-891a-5p in colon cancer tissue was significantly increased compared with that in normal colon epithelial cells (P<0.05). Compared with the group B, protein expressions of miR-891a-5p, Cyclin D1, N-cadherin and Vimentin were decreased in the group C, and cell proliferation rate, migration and invasion number and protein expressions of C-caspase-3, CPEB1 and E-cadherin and apoptosis rate were increased (P<0.05). Compared with the group D, the protein expressions of miR-891a-5p, Cyclin D1, N-cadherin and Vimentin, proliferation rate, migration and invasion number of cells were increased in the group E, while the protein expressions of C-caspase-3, CPEB1 and E-cadherin and apoptosis rate were decreased (P<0.05). There were six consecutive complementary binding sites between miR-891a-5p and CPEB1 at the 3 'UTR. Compared with the group F, the fluorescence activity and CPEB1 protein expression of WT-CPEB1 were decreased in the group G. Compared with the group B, the fluorescence activity of WT-CPEB1 was increased in the group C (P<0.05). Conclusion miR-891a-5p can promote the proliferation, migration and invasion of colon cancer cells and inhibit apoptosis, which may be related to the targeted inhibition of CPEB1.

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Ferrostatin-1 delay D-gal induced cardiomyocyte senescence by inhibiting ferroptosis

XIONG Xicheng, WANG Yiping, WANG Gang, ZHANG Tian, BAO Yali, Di na·AINIWAER, SUN Zhan

2023, 51 (1): 19-23. doi: 10.11958/20220818

Abstract ( 727 )

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Objective To investigate the existence of ferroptosis in cardiomyocyte senescence induced by D-galactose (D-gal) and whether ferroptosis inhibitor ferrostatin-1 (Fer-1) can delay cardiomyocyte senescence. Methods Cardiomyocyte injury was induced by D-gal (0, 5, 10, 20, 40, 80 and 100 g/L) to simulate cardiac aging model. After cells were treated with different concentrations of D-gal for 24 hours, the cell viability was detected by MTT method to determine the concentration of D-gal in the follow-up experiment. The cells were divided into the control group, the D-gal group and the Fer-1 group. The cell viability was detected by MTT method. The level of intracellular reactive oxygen species (ROS) was detected by DCFH-DA method. The intracellular glutathione (GSH) content was detected by microplate method. Intracellular malondialdehyde (MDA) content was detected by thiobarbituric acid method. The protein expression levels of SLC7A11, GPx4 and P53 were detected by Western blot assay. The activity of intracellular β-galactosidase (β-GAL) was detected by microassay. Results MTT assay showed that the cell viability decreased with the increase of D-gal concentration (P<0.05). The concentration of D-gal was 20 g/L in subsequent experiments. Compared with the D-gal group, the cell viability was increased in the Fer-1 group (P<0.05). Compared with the control group, ROS and MDA contents, P53 protein expression level and β -GAL activity were increased in the D-gal group, while GSH content, SLC7A11 and GPx4 protein expression levels were decreased (P<0.05). Compared with the D-gal group, ROS and MDA contents, P53 protein expression level and β -GAL activity were decreased in the Fer-1 group, while GSH, SLC7A11 and GPx4 protein expressions were increased (P<0.05). Conclusion Ferroptosis occurs during D-gal-induced cell senescence, and Fer-1 can inhibit iron death and delay myocardial senescence.

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The effect and mechanism of miR-15b gene interference on cerebral ischemia-reperfusion injury

SONG Zhengfeng, LIU Yuanyuan, QI Peng, TAN Xianxing, MA Lei

2023, 51 (1): 24-29. doi: 10.11958/20220433

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Objective To investigate the effect of microRNA-15b (miR-15b) gene interference on cerebral ischemia-reperfusion injury and its mechanism. Methods Astrocytes from cerebral cortex of newborn Wistar rats within 24 hours of birth were cultured and identified. Oxygen glucose deprivation/recovery method was used to treat the simulated cerebral ischemia-reperfusion injury in vivo, which was recorded as the model group. miR-15b interfering adenovirus vector and negative control vector were constructed and transfected into cells of the above model group respectively, which were recorded as the overexpression group, the silence group, the overexpression control group and the silence control group. Astrocytes from the cerebral cortex without special treatment were used as the blank control group. After 24 hours, the morphological changes of cells were observed by inverted phase contrast microscope in each group. The cell viability was detected by MTT assay, and the cell viability was detected by lactate dehydrogenase (LDH) leakage rate. The apoptosis rate was detected by flow cytometry. Real-time quantitative polymerase chain reaction was used to detect the expression levels of miR-15b, blymphoma-2 (Bcl-2), cysteine protease-3 (Caspase-3) and cysteine protease-9 (Caspase-9) messenger RNA (mRNA). The expressions of Bcl-2, Caspase-3 and Caspase-9 protein levels were detected by Western blot assay. Luciferase reporter gene experiment verified that miR-15b could target and regulate Bcl-2. Results Compared with the control group, the adherent cells decreased in the blank group, the overexpression control group and the silence control group, cells shrunk and became round, and the distribution was loose, cell viability and Bcl-2 mRNA and protein expression decreased, cell viability and apoptosis rate, and the expression levels of miR-15b, Caspase-3, Caspase-9 mRNA and protein increased (P<0.05). Compared with the blank group and the silencing control group, the number of adherent cells in the overexpression group decreased and was more sparsely distributed. It could be seen that cells floated in the culture medium, the cell survival rate and the expression of Bcl-2 mRNA and protein in the overexpression group decreased, the cell viability and apoptosis rate, and the expression of miR-15b, the expressions of Caspase-3, Caspase-9 mRNA and protein increased (P<0.05). Compared with the blank group and the silenced control group, the adherent cells in the silenced group increased, but still lower than the control group. The cells shrank slightly, the cell survival rate and the expression of Bcl-2 mRNA and protein increased in the silencing group, the cell viability and apoptosis rate, and the expression of miR-15b, the expressions of Caspase-3, Caspase-9 mRNA and protein decreased (P<0.05). Results of luciferase reporter gene experiments confirmed that miR-15b can target and regulate Bcl-2. Conclusion Overexpression of miR-15b can aggravate cerebral ischemia-reperfusion injury, presumably through the mitochondrial pathway to induce the deformation and apoptosis of damaged nerve cells.

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Effects of autonomic nervous regulation on myocardial structural remodeling, electrical remodeling and fibrosis in rats with ejection fraction preserved heart failure based on calcium overload

PENG Ming, LI Yukai, WANG Lan, HUANG Liang, CHENG Zhong, XIAO Jie

2023, 51 (1): 30-34. doi: 10.11958/20220897

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Objective To explore the effects of autonomic nervous regulation on structural remodeling, electrical remodeling and fibrosis in rats with ejection fraction preserved heart failure (HFpEF). Methods A total of 44 SPF grade 12-week-old SD rats were selected, 10 of them were selected as the control group, and the remaining 34 rats were used to establish the HFpEF model. The rat HFpEF model was established by ligation of abdominal aorta-inferior vena cava fistula. Among them, 30 rats were successfully modeled, and 4 rats died. The successfully established rats were divided into 3 groups with 10 rats in each group: the model group, the autonomic nerve regulation group and the autonomic nerve regulation+ acetylcholine M₂ receptor antagonist group. The autonomic nerve regulation group received percutaneous auricular marginal vagus nerve stimulation on the basis treatment of the model group. In the autonomic regulation + acetylcholine M₂ receptor antagonist group, mesotramine (0.5 mg/kg) was injected into the caudal vein every day on the basis treatment of the autonomic nervous regulation group. Left ventricular posterior wall end-diastolic dimension (LVPWD-D), interventricular ventricular septum end-diastolic dimension (IVS-D) and the peak value of mitral flow velocity in the early diastole/peak value of mitral flow velocity in late diastole (E/A) were measured by echocardiography.The effective refractory period (ERP) and monophasic action potential duration (MAPD) of rat hearts were obtained by cardiac electrophysiological stimulator in each group. The value of NT-proBNP was detected by ELISA. The disordered arrangement of cardiomyocytes and the infiltration of inflammatory cells were observed by HE staining. The degree of myocardial fibrosis was observed by Masson staining. The expression levels of matrix metalloproteinase-9 (MMP-9), tissue inhibitor of metalloproteinase-1 (TIMP-1), sarcoplasmic reticulum Ca²⁺-ATP enzyme (SERCA2a) and its regulatory protein phosphoprotein (PLB) mRNA and protein were detected by RT-PCR and Western blot assay. Results HE staining showed that the arrangement of cardiomyocytes was disordered, the intercellular space was not obvious enough, accompanied by the infiltration of inflammatory cells in the model group. Masson staining showed that the myocardial fibers were disordered, and a large number of collagen fibers were produced in the model group. The above pathological changes were significantly less in the autonomic nervous regulation group than those in the model group. The above pathological changes were not significantly improved in the autonomic nervous regulation + acetylcholine M₂ receptor antagonist group compared with those in the model group. Compared with the control group, the values of NT-proBNP, LVPWD-D, IVS-D, ERP, MAPD, MMP-9 mRNA and protein, PLB mRNA and protein were all increased in the model group, and levels of E/A, TIMP-1 mRNA and protein, SERCA2a mRNA and protein decreased significantly (P<0.05). Compared with the model group, values of NT-proBNP, LVPWD-D, IVS-D, ERP, MAPD, MMP-9 mRNA and protein, PLB mRNA and protein decreased in the autonomic nerve regulation group, and levels of E/A, TIMP-1 mRNA and protein, SERCA2a mRNA and protein were all increased (P<0.05). There were no significant differences in the above-mentioned indexes between the autonomic nervous regulation + acetylcholine M₂receptor antagonist group and the model group. Conclusion Autonomic nervous regulation may reduce structural remodeling, electrical remodeling and myocardial fibrosis in HFpEF model rats and improve the prognosis of HFpEF through reducing the level of calcium overload.

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ANA-12 relieves oxaliplatin-induced chemotherapy pain in rats by targetly inhibiting BDNF/TrkB signal

ZHAO Jiajia, WAN Wenjun, YANG Heyu, XIE Min, LIU Ling

2023, 51 (1): 35-40. doi: 10.11958/20220518

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Objective To investigate the effect and mechanism of ANA-12 on relieving oxaliplatin (OXA) induced neuropathic pain by targeting brain-derived neurotrophic factor (BDNF)/tyrosine kinase receptor B (TrkB) signaling in rats. Methods Eighteen male SD rats were randomly divided into 3 groups according to table of random number: the control group, the OXA treated group and the OXA + ANA-12 group (OXA+ANA-12), with 6 rats in each group. The OXA group and the OXA + ANA-12 group received an intraperitoneal injection of OXA (4 mg/kg for 5 days) to construct a chemotherapeutic pain model. After the model was successfully established, ANA-12 (20 g/L) was intrathecally administered in the OXA + ANA-12 group. After administration, pain behavior tests of rats in each group were performed, and changes in number of spontaneous flinches and mechanical pain threshold were recorded. The infiltration of spinal inflammatory cells and changes in expression level of interleukin (IL)-1β, tumor necrosis factor (TNF)-α, ionized calcium binding adapter molecule 1 (Iba1), BDNF, TrkB and nuclear factor kappa B (NF-κB) were detected by HE staining, immunofluorescence and Western blot assay. Results Compared with the control group, behavioral analysis showed that continuous injection of OXA significantly induced pain hyperalgesia and increased number of spontaneous flinches. Compared with the OXA group, intrathecal injection of ANA-12 significantly decreased the number of spontaneous flinches and increased mechanical pain threshold of rats. Morphological and protein expression analysis showed that OXA administration induced spinal inflammation, up-regulated BDNF/TrkB signaling and increased the expression levels of IL-1β, TNF-α, Iba1 and NF-κB compared with those of the control group. Compared with the OXA group, ANA-12 treatment significantly inhibited BDNF/TrkB signaling and down-regulated the expression levels of IL-1β, TNF-α, Iba1 and NF-κB. Conclusion Intrathecal administration of ANA-12 inhibits spinal inflammation and relieves chemotherapy pain by blocking BDNF/TrkB signal.

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The effect of halofuginone on cardiopulmonary function and its mechanism in rats with high-altitude pulmonary hypertension

WANG Jiangtao, MA Bohua, SHEN Huihua, HE Jia, YIN Dongfeng, WANG Rui, LI Wu

2023, 51 (1): 41-44. doi: 10.11958/20220965

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Objective To investigate the effect of halofuginone (HF) on the cardiopulmonary function of rats with high-altitude pulmonary hypertension (HAPH) and its mechanism. Methods Forty male SD rats were selected according to random number table method and divided into the model group, the low-dose, the medium-dose and the high-dose groups, with 10 rats in each group. HAPH rat model was constructed in hypobaric hypoxia chamber simulating altitude of 6 000 m. The other 10 were fed outside the cabin as the control group. Low, medium and high dose groups were given HF (0.5, 1 and 2 mg/kg, respectively). The model group and the control group were given distilled water, once a day, for consecutive 4 weeks. The mean pulmonary artery pressure (mPAP) was measured. The right ventricular hypertrophy index (RVHI) was calculated, and the degree of pulmonary vascular remodeling was assessed by hematoxylin-eosin (HE) staining. The levels of hypoxia inducible factor-1α (HIF-1α), vascular endothelial growth factor (VEGF) and endothelin-1 (ET-1) in serum and lung tissue of rats were detected by enzyme-linked immunosorbent assay (ELISA). Results Compared with the control group, the mPAP and RVHI were significantly increased in the model group (P<0.05). The small pulmonary artery wall was significantly thickened and the lumen was narrowed, and levels of HIF-1α, VEGF and ET-1 in serum and lung tissue were significantly increased (P<0.05). Compared with the model group, the mPAP and RVHI were significantly decreased in the low, medium and high dose groups (P<0.05). The degree of pulmonary vascular remodeling was significantly alleviated in the medium and high dose groups, and levels of HIF-1α, VEGF and ET-1 in serum and lung tissue were significantly decreased, while levels of VEGF and ET-1 in lung tissue were only decreased in the low-dose group (P<0.05). Compared with the low-dose group, the mPAP, VEGF and ET-1 in lung tissue were significantly decreased in the medium and high dose groups (P<0.05). Compared with the medium-dose group, there was no significant difference in each index in the high-dose group. Conclusion HF can reduce the mean pulmonary artery pressure, relieve right ventricular hypertrophy and improve pulmonary vascular remodeling in HAPH rats, and its mechanism may be related to the inhibition of HIF-1α, VEGF and ET-1 expressions.

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The expression of miR-149-3p in heart tissue of newborn mice with congenital heart disease and its effect on differentiation of P19 cells

HU Danhui, LUO Huichen, LIU Haiying

2023, 51 (1): 45-49. doi: 10.11958/20220707

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Objective To explore the expression of miR-149-3p in heart tissues of newborn mice with congenital heart disease (CHD) and its effect on differentiation of mouse embryonic teratoma P19 cells. Methods The newborn mouse model of ventricular septal defect CHD was established. On the 19th day of pregnancy, HE staining was used to observe pathological changes of fetal heart. Then qPCR was used to detect the expression levels of miR-149-3p and heat shock protein family B in fetal mouse heart tissue member 6 (HSPB6) mRNA in heart tissue of fetal mice. Dimethyl sulfoxide (DMSO) was used to induce P19 cells to differentiate into cardiomyocytes, and cells on the 0 th, 5 th and 10 th days of differentiation were collected. qPCR was used to detect markers of myocardial differentiation GATA4, cTnT, the mRNA expression level of atrial natriuretic peptide (ANP) and HSPB6 mRNA and miR-149-3p expression levels during the induction of differentiation. P19 cells were infected with miR-149-3p overexpression lentivirus and empty lentivirus. After induction for 10 days by DMSO, the expression of cTnI protein in cells was detected by immunofluorescence staining. mRNA expression levels of myocardial differentiation markers were detected by qPCR. The dual-luciferase reporter gene experiment was used to verify the targeting relationship between miR-149-3p and HSPB6. Results Compared with the normal group, the newborn mice in the CHD group developed cardiac ventricular septal defect, with high expression of miR-149-3p and low expression of HSPB6 in cardiac tissue (P<0.01). During the differentiation of P19 cells into cardiomyocytes, the expression levels of GATA4, cTnT, ANP and HSPB6 mRNA were gradually increased, while the expression level of miR-149-3p was gradually decreased (P<0.05). After 10 days of differentiation, the overexpression of miR-149-3p significantly reduced the differentiation rate of cardiomyocytes, and down-regulated the mRNA expression levels of GATA4, cTnT and ANP (P<0.01). The dual luciferase reporter gene experiment confirmed that miR-149-3p could target the expression of HSPB6. Conclusion miR-149-3p might inhibit the differentiation of P19 cells into cardiomyocytes through targeted down- regulation of HSPB6.

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The relationship between STAT4 rs7574865 single nucleotide polymorphism and ANCA-associated vasculitis

CAO Yueqi, XUE Chao

2023, 51 (1): 50-53. doi: 10.11958/20220821

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Objective To explore the association of STAT4 rs7574865 single nucleotide polymorphism with antineutrophil cytoplasmic autoantibody associated vasculitides (AAV). Methods A total of 145 patients with AAV (the AAV group) and 216 healthy controls (the control group) were included in this study. Whether the patients were accompanied by fever, proteinuria, hematuria and hemoptysis were collected. Multiplex PCR combined with high-throughput sequencing (MPCR-HTS) was used for genotyping. Genotypes, alleles and genotypes in different genetic models were compared between the two groups and between each clinical symptom subgroup. Logistic regression analysis was used to assess the relationship between genotype and risk of clinical symptoms. Results STAT4 rs7574865 genotype, allele and genotype distribution under different genetic models showed no significant difference between the two groups. There were no significant differences in genotype and allele distribution between different subgroups. In recessive model, the proportion of TT genotypes was lower in the hematuria group than that in the no hematuria group (P<0.05). Logistic regression analysis showed that TT genotype was not associated with the risk of hematuria. Conclusion STAT4 rs7574865 single nucleotide polymorphism was not associated with AAV susceptibility and clinical symptoms of fever, proteinuria, hematuria and hemoptysis.

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Clinical observation of oral glucocorticoid combined with intraocular pressure reduction in the treatment of patients with pathological myopia complicated with choroiditis

XIAO Bowen, CAO Wei, LIU Lijing, ZHAO Xiang, WANG Xin

2023, 51 (1): 54-57. doi: 10.11958/20220588

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Objective To observe the effect of oral glucocorticoid combined with ocular hypotensive drugs on the development of myopia in patients with pathologic myopia (PM) complicated with multifocal choroiditis (MFC). Methods A total of 56 PM patients (76 eyes) with MFC were collected, and they were treated with moderate-dose glucocorticoid (prednisone acetate) orally, with an average course of 13.5 weeks. At the same time, they were combined with brinzolamide eye drops for 24 months. Axial length (AL), spherical equivalent (SE), intraocular pressure, best corrected visual acuity (BCVA) and mean sensitivity of visual field ( mean sensitivity, MS) were followed-up and recorded at 3 months, 6 months, 12 months, 18 months and 24 months. Results After treatment, the color image of the fundus showed that the lesion area was slightly smaller than that before treatment, and the leakage of FFA fluorescein was improved compared with that before treatment. Compared with before treatment, there were no significant differences in AL, SE and intraocular pressure only at 3 months after treatment, and there were no significant differences in AL and SE at 24 months after treatment. Compared with before treatment and 3 months after treatment, there were significant differences in each index at 6, 12 and 18 months after treatment (P<0.05). There were significant differences in BCVA and visual field MS at different time points between 3, 6 and 24 months after treatment (P<0.05). At 3 and 24 months after treatment, there was no correlation between AL and intraocular pressure and SE. At 6, 12 and 18 months after treatment, AL was positively correlated with intraocular pressure and negatively correlated with SE (P<0.01). Conclusion Oral glucocorticoid combined with ocular hypotensive drugs to treat PM with MFC can control the growth of PM AL and myopia diopter, and effectively improve visual function.

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Effects of DNase I combined with Nd:YAP laser on the removal of enterococcus faecalis biofilm in root canals

YE Jing, LIU Qing, WEN Te, DU Rui, WANG Yuan, MA Ben, NI Na

2023, 51 (1): 58-61. doi: 10.11958/20220653

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Objective To study the effect of DNaseⅠ combined with Nd: YAP laser on the removal of enterococcus faecalis (E.f) biofilm in root canals. Methods The E.f infection model on dentin surface was established and observed by scanning electron microscope (SEM). E.f biofilm samples cultured for 48 h were divided into 5 groups by random number table method, with 5 samples in each group. The group A was treated with 0.5%NaOCl, the group B was treated with 0.5%NaOCl+Nd: YAP laser, the group C was treated with DNaseⅠ+ 0.5%NaOCl, the group D was treated with DNaseⅠ+ 0.5%NaOCl+Nd: YAP laser and the group E was treated with 0.9% normal saline. The adhesion of E.f on dentin surface was observed by laser confocal microscope (CLSM), and the amount of green fluorescence was counted, which was the amount of viable bacteria. Results SEM observation showed the dentin tubule opening on the dentin surface. E.f adhesion was observed on the dentin surface, and bacterial adhesion was observed in the deep layer of dentin tubule. CLSM observation showed that living bacteria with green fluorescence were scattered on the dentin surface in the group A, the group B and the group C. Less green fluorescence was observed in the group D. Green fluorescent live bacteria covered all the sample surface in the group E. The amount of bacteria in the group D was significantly less than that in the groups A and the groups E, and the amount of bacteria in the group C was significantly less than that in the group E (P<0.01). Conclusion Compared with 0.5% NaOCl alone, DNaseⅠ combined with Nd: YAP laser and 0.5% NaOCl can treat E.f biofilms more effectively.

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Relationship between serum uric acid level and abnormal renal function in adults

WANG Jianxiong, LI Chunjun, ZHANG Li, HE Jiangshan, GUO Pei, ZHANG Minying, ZHANG Mianzhi

2023, 51 (1): 62-68. doi: 10.11958/20220663

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Objective To explore the relationship between serum uric acid (SUA) levels and renal function decline in adults. Methods A total of 12 138 normal physical examination subjects were selected as research subjects. General data of subjects were collected by questionnaire, and serum creatinine (Scr), SUA and fasting blood glucose were measured. Subjects were divided into two groups based on estimated glomerular filtration rate (eGFR): the abnormal renal function group (1 607 cases) and the normal renal function group (10 531 cases). Men and women of subjects were divided into the hyperuricemia group and the normal uric acid group according to 420 μmol/L and 360 μmol/L uric acid levels, respectively. The normal uric acid group was then divided into four sub-groups according to SUA quartile. Trends in the proportion of patients with abnormal renal dysfunction with SUA levels were analyzed in different genders. Logistic regression was used to analyze the influence of SUA level and renal function in subgroups with different gender, age, education level, exercise, hypertension and diabetes. Results In men and women, with the increase of SUA level, the proportion of SUA quartile groups and hyperuricemia patients in the renal dysfunction group showed an increasing trend (Z _male =6.411; Z _female =23.800, P<0.01). Logistic regression results showed that compared with the first quartile group, the second, third, and fourth quartile groups and hyperuricemia group were all risk factors of renal dysfunction (P<0.05). There was a significant dose-response relationship between SUA and renal dysfunction in both men and women (P _{for trend}<0.01). The relationship was statistically significant across subgroups of age, education, physical activity, hypertension and diabetes. Conclusion Elevated SUA are related with an increased risk of renal dysfunction in a dose-response relationship.

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The correlation between the level of reticulin 1C in peripheral blood mononuclear cells and the efficacy of neoadjuvant chemotherapy for triple negative breast cancer

ZHANG Manli, LIU Weiwei, WANG Lijuan, LI Weidong

2023, 51 (1): 69-73. doi: 10.11958/20220641

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Objective To explore the relationship between the expression level of reticulin 1C (RTN-1C) in peripheral blood mononuclear cells of patients with triple negative breast cancer (TNBC) and the efficacy of neoadjuvant chemotherapy. Methods A total of 154 TNBC patients were selected as the study subjects. Patients were divided into the complete remission group (n=47) and the incomplete remission group (n=107) according to the efficacy of neoadjuvant chemotherapy. Data of age, menstrual status, type of pathology, TNM stage, histological grade, Ki-67 expression >30% and tumour diameter were collected from all subjects. Fasting elbow venous blood samples were collected from TNBC patients before chemotherapy (T₀), 7 days after chemotherapy (T₁), 14 days after chemotherapy (T₂) and 21 days after chemotherapy (T₃). Peripheral blood mononuclear cells were isolated by Ficoll density gradient centrifugation. The expression levels of RTN-1C in mononuclear cells were detected by Western blot assay. The receiver operating characteristic curve (ROC) was used to evaluate the efficacy of RTN-1C in determining neoadjuvant chemotherapy efficacy. Logistic regression was used to analyse risk factors for neoadjuvant chemotherapy efficacy. Nomogram regression models were constructed to predict complete remission of pathology after neoadjuvant chemotherapy, and consistency index (C-index), calibration curves and decision tree curves were used to assess model efficacy. Results The relative expression levels of RTN-1C were lower at T₁ and T₃ in the complete remission group than those in the incomplete remission group (P<0.05). The relative expression levels of RTN-1C at T₀, T₁ and T₂ decreased over time in the 2 groups (P<0.01). The area under the ROC curve (AUC) of T₃-RTN-1C was higher than T₀-RTN-1C, T₁-RTN-1C and T₂-RTN-1C in judging pathological complete remission after neoadjuvant chemotherapy (P<0.01). Logistic regression analysis showed that T₃-RTN-1C>0.91 (OR=12.178, 95%CI: 4.796-30.924), N1 or N2 (OR=2.180, 95%CI: 1.100-4.322) and histological grade Ⅲ (OR=3.609, 95%CI: 1.453-8.969) were independent risk factors for pathological incomplete remission after neoadjuvant chemotherapy (P<0.05). Model A (constructed by N stage, histological grade and T₃-RTN-1C) had a high degree of coincidence between the fitting curve and the ideal curve, while model B (constructed by N stage and histological grade) had a poor degree of coincidence between the fitting curve and the ideal curve. The C-index of model A and model B were 0.866 and 0.772, respectively. When the threshold probability was higher than 0.40, the net benefit of model A in judging pathological complete response after neoadjuvant chemotherapy was higher than that of model B. Conclusion The model constructed by N stage, histological grade and T₃-RTN-1C has a high degree of differentiation, accuracy and clinical application value in judging pathological complete response after neoadjuvant chemotherapy.

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Predictive value of serum FGF-23 in recurrence of patients with atrial fibrillation after radiofrequency ablation

HUO Liwei, LIU Jun, ZHENG Binbin, BI Xuena

2023, 51 (1): 74-77. doi: 10.11958/20220317

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Objective To explore the predictive value of fibroblast growth factor-23 (FGF-23) in recurrence of patients with atrial fibrillation (AF) after radiofrequency ablation. Methods A total of 132 AF patients who received radiofrequency ablation were selected. According to the recurrence 12 months after radiofrequency ablation, patients were divided into the recurrence group (41 cases) and the non-recurrence group (91 cases). In addition, according to the expression level of serum FGF-23, patients were divided into the FGF-23 high expression group (56 cases) and the FGF-23 low expression group (76 cases). The parameters such as left atrial diameter (LAD) were detected by color Doppler ultrasound diagnostic instrument, and the serum level of FGF-23 was detected by enzyme-linked immunosorbent assay kit. Results Compared with the non-recurrent group, the proportion of persistent AF, LAD and serum FGF-23 levels were higher in the recurrence group (P<0.05). Persistent AF, excessive LAD and high serum FGF-23 level were risk factors for recurrence in AF patients after radiofrequency ablation (P<0.05). Receiver operating characteristic curve (ROC) analysis showed that the areas under the curve of serum FGF-23, LAD and their combined detection to predict the recurrence of AF patients after radiofrequency ablation were 0.770 (95%CI: 0.681-0.859), 0.743 (95%CI: 0.651-0.836) and 0.828 (95%CI: 0.752-0.904) respectively. The recurrence rate after radiofrequency ablation was significantly higher in the FGF-23 high expression group (FGF-23≥224.42 μg/L, 56 cases) than that of the FGF-23 low expression group (FGF-23<224.42 μg/L, 76 cases, 44.64% vs. 21.05%, χ²=8.379, P<0.05). Conclusion High serum FGF-23 level is a risk factor for recurrence in AF patients after radiofrequency ablation, and it has high predictive value for recurrence after radiofrequency ablation in patients with AF.

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Distribution characteristics of bacteria isolated from lower respiratory tract secretions of premature infants with severe bronchopulmonary dysplasia

GU Mingxiao, LIU Xuancheng, LI Yue, SHAN Ruobing

2023, 51 (1): 78-80. doi: 10.11958/20220841

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Objective To analyze the characteristics of bacteria isolated from lower respiratory tract in premature infants with severe bronchopulmonary dysplasia (BPD), and to explore the correlation between the bacteria and BPD. Methods A total of 185 preterm infants diagnosed as BPD in the neonatal intensive care unit (NICU) with invasive mechanical ventilation ≥ 72 h were enrolled in the study. Patients were divided into the mild moderate BPD group (n=124) and the severe BPD group (n=61) according to the severity of the disease. The general clinical data such as birth weight and gestational age were collected, as well as the treatment process indicators of neonatal diseases such as necrotizing enterocolitis, sepsis and ventilator-associated pneumonia (VAP). Respiratory aspirate fluid (TAF) samples from 2 groups were collected for pathogen culture and statistically analyzed. Results Compared with the mild moderate group, birth weight and gestational age were lower in the severe group (P<0.05), and the proportion of children with gram-negative bacteria, Klebsiella pneumoniae, Pseudomonas aeruginosa and Corynebacterium detected by TAF were increased (P<0.05). Conclusion Changes of lower respiratory tract flora in preterm infants may be an important factor in the development of BPD, and the proportion of Klebsiella pneumoniae, Pseudomonas aeruginosa and Corynebacterium detected in TAF of children with severe BPD is higher.

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Clinical study on the evaluation of dynamic endometrial morphology and receptivity for IVF-ET pregnancy outcomes of patients with hydrosalpinx infertility

LIANG Ting, YANG Jingya, LIANG Qin, ZHOU Cuijuan

2023, 51 (1): 81-85. doi: 10.11958/20220414

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Objective To explore the evaluation of dynamic endometrial morphology and receptivity for pregnancy outcomes of in vitro fertilization-embryo transfer (IVF-ET) in patients with hydrosalpinx infertility. Methods A total of 99 patients with hydrosalpinx infertility were enrolled. All were given antagonist regimens for assisted reproduction. According to the first IVF-ET pregnancy outcomes, patients were divided into the pregnancy group (43 cases) and the non-pregnancy group (56 cases). The endometrial morphology and hemodynamic indexes were compared between the two groups. The specimens of endometrial biopsy in implantation window of oocyte retrieval period were collected to detect the expressions of homeobox gene A10 (HOXA10) and integrin α_vβ3 by immunohistochemical staining. Results There were no significant differences in uterine position and endometrial morphology between the two groups (P>0.05). On the start day of COH cycle (D1), uterine artery PI was lower in the pregnancy group than that in the non-pregnancy group. On the day of HCG injection (D2), endometrial thickness was greater in the pregnancy group than that in the non-pregnancy group, and uterine artery PI and RI were lower in the pregnancy group than those in the non-pregnancy group (P<0.01). The expression levels of HOXA10 and integrin α_vβ3 in endometrial tissue were significantly higher in the pregnancy group than those in the non-pregnancy group (P<0.05). On D2, the expression of RI in uterine artery, HOXA10 and integrin α_vβ3 in endometrial tissue were independent factors influencing pregnancy outcomes of IVF-ET (P<0.05). The expression levels of HOXA10 and integrin α_vβ3 in endometrial tissue were of certain predictive value for IVF-ET pregnancy, and the area under the curve (AUC) were 0.778 (95%CI: 0.681-0.874) and 0.779 (95%CI: 0.685-0.873), respectively. Conclusion The endometrial receptivity indexes HOXA10 and α_vβ3 have a positive predictive effect on pregnancy outcomes of IVF-ET in patients with hydrosalpinx infertility.

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Analysis of long-term effect and risk factors of modified TROPIS on postoperative anal function in patients with high anal fistula

HOU Yi, ZHENG Xueping, TAN Yanyan, ZHOU Feng, HUANGFU Shaohua, XU Dachao, WANG Yan

2023, 51 (1): 86-90. doi: 10.11958/20220756

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Objective To evaluate the long-term effect of modified TROPIS on postoperative anal function in patients with high anal fistula, and analyze the risk factors influencing postoperative anal function. Methods A retrospective case-control study was used in this study. A total of 181 patients with high anal fistula who underwent surgical treatment were included. Patients were divided into the experimental group (n = 84) and the control group (n = 97) according to the surgical method. The experimental group was treated with modified TROPIS, and the control group was treated with incision-thread-drawing procedure. Follow-up period was 12-36 months (median, 21 months). The general condition, cure rate and anal function were compared between the two groups. Logistic regression analysis was used to analyze risk factors influencing postoperative anal function, and the area under the ROC curve (AUC) and the Hosmer-Lemeshow test were used to evaluate the predictive efficacy. Results The relapse rate was 8.33% in the experimental group, which was significantly lower than 20.62% in the control group. The total score of postoperative anal gas control, lifestyle and Wexner incontinence were significantly higher after surgery compared with those before surgery (P<0.01), but each score was significantly lower than that in the control group (P<0.01). Postoperative anal function declined in 79 patients. Logistic regression analysis showed that body mass index (OR=4.579), horseshoe anal fistula (OR=158.011), number of fistulas (OR=8.789), lesion involving external sphincter (OR=6.883) and surgical method (OR=7.082) were independent risk factors for postoperative anal function decline in patients with high anal fistula. Combined with each independent risk factor, the AUC of overall predictive probability was 0.903 (95%CI: 0.859-0.947, P<0.01). Hosmer-Lemeshow test demonstrated that χ²=5.603, P=0.587, indicating that the combined prediction of each independent risk factor had high discrimination and accuracy. Conclusion Compared with the incision-thread-drawing procedure, modified TROPIS procedure is safer and more effective in the treatment of high anal fistula. However, this surgery still leads to long-term postoperative anal function decline. For patients with obesity and complicated fistula, anal sphincter should be protected as much as possible during surgery.

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Relationship between irisin and newly diagnosed type 2 diabetes mellitus

TAN Anjun, YANG Jingjing, LI Tianrong

2023, 51 (1): 91-94. doi: 10.11958/20220376

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Objective To explored the relationship between irisin and newly diagnosed type 2 diabetes mellitus (T2DM). Methods Forty newly diagnosed T2DM patients were collected as the T2DM group. During the same period, 40 healthy subjects in the physical examination center of the hospital were collected as the control (NC) group. Clinical indexes, metabolic indexes and serum concentrations of inflammatory cytokines interleukin-1β (IL-1β), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α) and irisin were detected in the two groups. The differences of above indexes were analysed in the two groups. The correlation between irisin and the above indexes was analyzed. Logistic regression was used to analyze the relationship of irisin and newly diagnosed T2DM. Results Compared with the NC group, serum levels of cytokines IL-1β, IL-6, TNF-α and IFN-γ were significantly increased in the T2DM group (P<0.05), and the serum level of irisin was significantly decreased (P<0.05). The serum irisin was negatively correlated with FPG, 2 h PG, HbA1c, HOMA-IR, IL-1β and IFN-γ (P<0.05). The increased level of irisin was a protective factor for T2DM (P<0.01). Conclusion The serum level of irisin decreases in newly diagnosed T2DM patients. Irisin is closely related to glucose metabolism, insulin resistance and inflammation, which may have some protective effects on T2DM.

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Construction of a multiple risk factor prediction model for colorectal adenoma in patients with type 2 diabetes mellitus

YANG Chao, ZHANG Zhiyong

2023, 51 (1): 95-99. doi: 10.11958/20220636

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Objective To investigate independent influencing factors of colorectal adenomas (CRA) in patients with type 2 diabetes mellitus (T2DM) and to establish the risk prediction model. Methods Patients with T2DM undergoing colonoscopy were divided into the adenoma group (n=145, pathologically diagnosed as CRA) and the control group (n=174, without abnormal colorectal mucosa). Age, sex, body mass index (BMI) and diabetes course, history of hypertension, diabetic peripheral neuropathy, fatty liver, gastrointestinal symptoms, metformin and insulin use in recent 5 years, Helicobacter pylori (Hp) infection, smoking history, drinking history and fecal occult blood were compared between the two groups. Laboratory data were also compared between the two groups, including tumor abnormal protein (TAP), glycosylated hemoglobin (HbA1c), insulin resistance index (HOMA-IR), serum total cholesterol (TC), triglyceride (TG), low density lipoprotein cholesterol (LDL-C), uric acid (UA) and vitamin D₃ levels. Multivariate Logistic regression was used to determine the independent risk factors for CRA in T2DM patients, and a new combined predictor was constructed (L_combined). The combined prediction value of L_combined was evaluated by ROC curve, and the fit degree was evaluated by Hosmer-Lemeshow test. Results In the adenoma group, age, BMI, male, history of hypertension, history of smoking, history of drinking, positive proportion of Hp, positive proportion of stool occult blood, HbA1c, TG, TAP and HOMA-IR were significantly higher than those in the control group. The proportion of metformin use and vitamin D₃ levels were lower in the adenoma group than those in the control group (P<0.05). Multivariate Logistic regression showed that the increased age, BMI, TAP, TG, smoking history, positive Hp and positive occult blood were independent risk factors for CRA in T2DM patients, while metformin was an independent protective factor (P<0.05). ROC curve showed that the AUC of L_combined was 0.815 (95%CI: 0.768-0.862, P<0.01), and Hosmer-Lemeshow test showed a good fit of the risk model (χ²=10.249, P=0.248). Conclusion The combined predictors of age, BMI, TAP, TG, smoking history, Hp infection, fecal occult blood and metformin use are effective in predicting the risk of colorectal adenoma in patients with T2DM.

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The influencing factors and clinical diagnostic value of dry eye syndromes

XING Jiannan, HE Wei

2023, 51 (1): 100-104. doi: 10.11958/20220589

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Objective To explore the influencing factors and clinical diagnostic value of dry eyes in patients with dry eye syndromes. Methods A total of 160 patients with dry eye were collected，including the mild dry eye group (n=51), the moderate dry eye group (n=61) and the severe dry eye group (n=48). Another 55 healthy subjects were selected as the normal control group. Data of the first tear film rupture time (NIBUTf), mean tear film rupture time (NIBUTav), tear film lipid layer thickness (LLT), lacrimal river height (LTMH) and conjunctival congestion score were detected by dry eye analyzer SLM-KD3. The ocular surface disease index (OSDI) scale score, corneal fluorescent staining score (FL) and tear secretion test (Slt) were performed in each group. Interleukin-17 (IL-17) and matrix metalloproteinase-9 (MMP-9) in tears were detected by enzyme-linked immunosorbent assay (ELISA). Univariate and multivariate Logistic regression were used to analyze influencing factors of dry eye. Results NIBUTf, NIBUTav, LLT, LTMH and SIt decreased in turn in the control group, the mild dry eye group, the moderate dry eye group and the severe dry eye group, while IL-17, MMP-9, OSDI and FL scores increased in turn (P<0.05). The score of conjunctival congestion was higher in the severe dry eye group than that in the control group and the mild dry eye group (P<0.05). Logistic regression analysis showed that higher levels of LLT and LTMH were protective factors for dry eyes, while higher levels of IL-17 and MMP-9 were risk factors for dry eyes. ROC analysis showed that when each individual index was diagnosed, the comprehensive advantage of LLT was more obvious, and the diagnostic specificity of IL-17 was higher, and the advantage of combined diagnosis of each index was the most obvious. Conclusion Higher levels of LLT and LTMH are protective factors for dry eyes, while higher levels of IL-17 and MMP-9 are risk factors for dry eyes. The combined detection of the four indexes is helpful in the diagnosis of dry eyes.

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Research progress of influencing mechanism of COVID-19 on intestinal microbiota and related intestinal mucosal barrier defects

SUN Qiang, SONG Weiliang

2023, 51 (1): 105-108. doi: 10.11958/20220608

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The interaction between intestinal microbiota and intestinal mucosal barrier plays an important role in maintaining the stability of intestinal environment. Respiratory syndrome coronavirus infection can directly or indirectly disrupt the diversity of intestinal microflora through different ways, and affect the structure and metabolism of intestinal mucosal barrier. The intestinal homeostasis and microenvironment appear to play a major role in the pathogenesis of coronavirus disease 2019 (COVID-19) and the enhancement of the systemic inflammatory responses, suggesting that intestinal microbiota and mucosal barrier can be used as important therapeutic targets to reverse the pathological process of these patients. Therefore, this paper is a review about the research progress on the mechanism of COVID-19 induced intestinal microbiota and related intestinal mucosal barrier defects.

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Research progress of interleukin 22 binding protein in inflammation related diseases

SHI Jingxiang, CAO Jisen, ZHANG Jinjuan, WANG Yijun

2023, 51 (1): 109-112. doi: 10.11958/20220999

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Interleukin 22 binding protein (IL-22BP) is a soluble receptor homolog that binds to interleukin 22 (IL-22) with greater affinity than the membrane spanning receptor. IL-22BP can play a role in maintaining immune homeostasis by inhibiting its expression and combining with IL-22. This review will focus on the production, regulation, typing of IL-22BP and its research progress in inflammatory related diseases, and explore the feasibility of it as a therapeutic target.

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