1.Dynamics of eosinophil infiltration and microglia activation in brain tissues of mice infected with Angiostrongylus cantonensis
Fanna WEI ; Renjie ZHANG ; Yahong HU ; Xiaoyu QIN ; Yunhai GUO ; Xiaojin MO ; Yan LU ; Jiahui SUN ; Yan ZHOU ; Jiatian GUO ; Peng SONG ; Yanhong CHU ; Bin XU ; Ting ZHANG ; Yuchun CAI ; Muxin CHEN
Chinese Journal of Schistosomiasis Control 2025;37(2):163-175
Objective To investigate the changes in eosinophil counts and the activation of microglial cells in the brain tissues of mice at different stages of Angiostrongylus cantonensis infection, and to examine the role of microglia in regulating the progression of angiostrongyliasis and unravel the possible molecular mechanisms. Methods Fifty BALB/c mice were randomly divided into the control group and the 7-d, 14-d, 21-day and 25-d infection groups, of 10 mice in each group. All mice in infection groups were infected with 30 stage III A. cantonensis larvae by gavage, and animals in the control group was given an equal amount of physiological saline. Five mice were collected from each of infection groups on days 7, 14, 21 d and 25 d post-infection, and 5 mice were collected from the control group on the day of oral gavage. The general and focal functional impairment was scored using the Clark scoring method to assess the degree of mouse neurological impairment. Five mice from each of infection groups were sacrificed on days 7, 14, 21 d and 25 d post-infection, and 5 mice from the control group were sacrificed on the day of oral gavage. Mouse brain tissues were sampled, and the pathological changes of brain tissues were dynamically observed using hematoxylin and eosin (HE) staining. Immunofluorescence staining with eosinophilic cationic protein (ECP) and ionized calcium binding adaptor molecule 1 (Iba1) was used to assess the degree of eosinophil infiltration and the counts of microglial cells in mouse brain tissues in each group, and the morphological parameters of microglial cells (skeleton analysis and fractal analysis) were quantified by using Image J software to determine the morphological changes of microglial cells. In addition, the expression of M1 microglia markers Fcγ receptor III (Fcgr3), Fcγ receptor IIb (Fcgr2b) and CD86 antigen (Cd86), M2 microglia markers Arginase 1 (Arg1), macrophage mannose receptor C-type 1 (Mrc1), chitinase-like 3 (Chil3), and phagocytosis genes myeloid cell triggering receptor expressed on myeloid cells 2 (Trem2), CD68 antigen (Cd68), and apolipoprotein E (Apoe) was quantified using real-time quantitative reverse transcription PCR (RT-qPCR) assay in the mouse cerebral cortex of mice post-infection. Results A large number of A. cantonensis larvae were seen on the mouse meninges surface post-infection, and many neuronal nuclei were crumpled and deeply stained, with a large number of bleeding points in the meninges. The median Clark scores of mouse general functional impairment were 0 (interquartile range, 0), 0 (interquartile range, 0.5), 6 (interquartile range, 1.0), 14 (interquartile range, 8.5) points and 20 (interquartile range, 9.0) points in the control group and the 7-d, 14-d, 21-d and 25-d groups, respectively (H = 22.45, P < 0.01), and the median Clark scores of mouse focal functional impairment were 0 (interquartile range, 0), 2 (interquartile range, 2.5), 7 (interquartile range, 3.0), 18 (interquartile range, 5.0) points and 25 (interquartile range, 6.5) points in the control group and the 7-d, 14-d, 21-d and 25-d groups, respectively (H = 22.72, P < 0.01). The mean scores of mice general and focal functional impairment were all higher in the infection groups than in the control group (all P values < 0.05). Immunofluorescence staining showed a significant difference in the eosinophil counts in mouse brain tissues among the five groups (F = 40.05, P < 0.000 1), and the eosinophil counts were significantly higher in mouse brain tissues in the 14-d (3.08 ± 0.78) and 21-d infection groups (5.97 ± 1.37) than in the control group (1.00 ± 0.28) (both P values < 0.05). Semi-quantitative analysis of microglia immunofluorescence showed a significant difference in the counts of microglial cells among the five groups (F = 17.66, P < 0.000 1), and higher Iba1 levels were detected in mouse brain tissues in 14-d (5.75 ± 1.28), 21-d (6.23 ± 1.89) and 25-d infection groups (3.70 ± 1.30) than in the control group (1.00 ± 0.30) (all P values < 0.05). Skeleton and fractal analyses showed that the branch length [(162.04 ± 34.10) μm vs. (395.37 ± 64.11) μm; t = 5.566, P < 0.05] and fractal dimension of microglial cells (1.30 ± 0.01 vs. 1.41 ± 0.03; t = 5.266, P < 0.05) were reduced in mouse brain tissues in the 21-d infection group relative to the control group. In addition, there were significant differences among the 5 groups in terms of M1 and M2 microglia markers Fcgr3 (F = 48.34, P < 0.05), Fcgr2b (F = 55.46, P < 0.05), Cd86 (F = 24.44, P < 0.05), Arg1 (F = 31.18, P < 0.05), Mrc1 (F = 15.42, P < 0.05) and Chil3 (F = 24.41, P < 0.05), as well as phagocytosis markers Trem2 (F = 21.19, P < 0.05), Cd68 (F = 43.95, P < 0.05) and Apoe (F = 7.12, P < 0.05) in mice brain tissues. Conclusions A. cantonensis infections may induce severe pathological injuries in mouse brain tissues that are characterized by massive eosinophil infiltration and persistent activation of microglia cells, thereby resulting in progressive deterioration of neurological functions.
2.Dynamic observation on capillarization of liver sinusoidal endothelial cells induced by Echinococcus multilocularis infection
Renjie ZHANG ; Jun XIE ; Fanna WEI ; Xiaojin MO ; Peng SONG ; Yuchun CAI ; Yan LU ; Jiahui SUN ; Yan ZHOU ; Lin LIN ; Ting ZHANG ; Muxin CHEN
Chinese Journal of Schistosomiasis Control 2024;36(1):34-43
Objective To investigate the capillarization of liver sinusoidal endothelial cells (LSECs) and its association with hepatic fibrosis during the development of alveolar echinococcosis, so as to provide the basis for unraveling the mechanisms underlying the role of LSEC in the development and prognosis of hepatic injuries and hepatic fibrosis caused by alveolar echinococcosis. Methods Forty C57BL/6 mice at ages of 6 to 8 weeks were randomly divided into a control group and 1-, 2- and 4-week infection groups, of 10 mice in each group. Each mouse in the infection groups was intraperitoneally injected with 2 000 Echinococcus multilocularis protoscoleces, while each mouse in the control group was given an equal volume of phosphate-buffered saline using the same method. All mice were sacrificed 1, 2 and 4 weeks post-infection and mouse livers were collected. The pathological changes of livers were observed using hematoxylin-eosin (HE) staining, and hepatic fibrosis was evaluated through semi-quantitative analysis of Masson’s trichrome staining-positive areas. The activation of hepatic stellate cells (HSCs) and extracellular matrix (ECM) deposition were examined using immunohistochemical staining of α-smooth muscle actin (α-SMA) and collagen type I alpha 1 (COL1A1), and the fenestrations on the surface of LSECs were observed using scanning electron microscopy. Primary LSECs were isolated from mouse livers, and the mRNA expression of LSEC marker genes Stabilin-1, Stabilin-2, Ehd3, CD209b, GATA4 and Maf was quantified using real-time fluorescence quantitative PCR (qPCR) assay. Results Destruction of local liver lobular structure was observed in mice 2 weeks post-infection with E. multilocularis protoscoleces, and hydatid cysts, which were surrounded by granulomatous tissues, were found in mouse livers 4 weeks post-infection. Semi-quantitative analysis of Masson’s trichrome staining showed a significant difference in the proportion of collagen fiber contents in mouse livers among the four groups (F = 26.060, P < 0.001), and a higher proportion of collagen fiber contents was detected in mouse livers in the 4-week infection group [(11.29 ± 2.58)%] than in the control group (P < 0.001). Immunohistochemical staining revealed activation of a few HSCs and ECM deposition in mouse livers 1 and 2 weeks post-infection, and abundant brown-yellow stained α-SMA and COL1A1 were deposited in the lesion areas in mouse livers 4 weeks post-infection, which spread to surrounding tissues. Semi-quantitative analysis revealed significant differences in α-SMA (F = 7.667, P < 0.05) and COL1A1 expression (F = 6.530, P < 0.05) in mouse levers among the four groups, with higher α-SMA [(7.13 ± 3.68)%] and COL1A1 expression [(13.18 ± 7.20)%] quantified in mouse livers in the 4-week infection group than in the control group (both P values < 0.05). Scanning electron microscopy revealed significant differences in the fenestration frequency (F = 37.730, P < 0.001) and porosity (F = 16.010, P < 0.001) on the surface of mouse LSECs among the four groups, and reduced fenestration frequency and porosity were observed in the 1-[(1.22 ± 0.48)/μm2 and [(3.05 ± 0.91)%] and 2-week infection groups [(3.47 ± 0.10)/μm2 and (7.57 ± 0.23)%] groups than in the control group (all P values < 0.001). There was a significant difference in the average fenestration diameter on the surface of mouse LSECs among the four groups (F = 15.330, P < 0.001), and larger average fenestration diameters were measured in the 1-[(180.80 ± 16.42) nm] and 2-week infection groups [(161.70 ± 3.85) nm] than in the control group (both P values < 0.05). In addition, there were significant differences among the four groups in terms of Stabilin-1 (F = 153.100, P < 0.001), Stabilin-2 (F = 57.010, P < 0.001), Ehd3 (F = 31.700, P < 0.001), CD209b (F = 177.400, P < 0.001), GATA4 (F = 17.740, P < 0.001), and Maf mRNA expression (F = 72.710, P < 0.001), and reduced mRNA expression of Stabilin-1, Stabilin-2, Ehd3, CD209b, GATA4 and Maf genes was quantified in three infection groups than in the control group (all P values < 0.001). Conclusions E. multilocularis infections may induce capillarization of LSECs in mice, and result in a reduction in the expression of functional and phenotypic marker genes of LSECs, and capillarization of LSECs occurs earlier than activation of HSC and development of hepatic fibrosis.
3.Pathological characteristics of false-positive lesions of prostate cancer on 68Ga-PSMA-11 PET/CT
Renjie LI ; Yao FU ; Shan PENG ; Fengjiao YANG ; Feng WANG ; Hongqian GUO ; Xuefeng QIU
Journal of Modern Urology 2024;29(11):988-992
[Objective] To investigate the pathological characteristics of false-positive lesions of prostate cancer on 68Ga-PSMA-11 PET/CT based on the pathology of whole mount specimens, in order to more accurately assess the degree of malignancy within the prostate tissue and avoid overdiagnosis and unnecessary treatment. [Methods] A total of 77 patients who underwent 68Ga-PSMA-11 PET/CT before radical prostatectomy in Nanjing Drum Tower Hospital during Jan.2018 and Dec.2022 were retrospectively analyzed.The pathology of whole mount specimens was detected.Two nuclear physicians examined all imaging plates without knowing the pathological results.Two pathological physicians completed all pathological diagnosis without knowing the imaging results.The pathological characteristics of false-positive lesions were determined by matching 68Ga-PSMA-11 PET/CT and pathological specimens.To analyze the pathological features of false-positive lesions, true-negative lesions were randomly delineated and defined.The pathological features of false-positive and true-negative lesions were analyzed and compared using Fisher exact test. [Results] After the imaging and pathological sections were matched, 21(16.3%) false-positive lesions were identified.The pathological characteristics of the 21 false-positive lesions were as follows: 16 (76.2%) simple atrophy with cyst formation, 3(14.3%) prostatic nodular hyperplasia, and 2(9.5%) inflammation.The pathological characteristics of 21 true-negative lesions were: 13(61.9%) normal glands, 5(23.8%) prostatic nodular hyperplasia and 3(14.3%) simple atrophy with cyst formation.Fisher exact test showed that the proportion of simple atrophy with cyst formation in the pathological features of false-positive lesions and true-negative lesions was statistically significant (76.2% vs.14.3%, P<0.001). [Conclusion] Simple atrophy with cyst formation may be a characteristic pathological type of the false-positive lesions of prostate cancer on 68Ga-PSMA-11 PET/CT.
4.Health risk assessment of dietary lead exposure among children in Jiading District
DONG Yuting ; PENG Hui ; WANG Xiaohui ; CHENG Renjie ; WANG Hexing
Journal of Preventive Medicine 2024;36(10):893-896
Objective:
To investigate the dietary lead exposure level among children aged 3 to 4 years in Jiading District, Shanghai Municipality, and assess the health risk caused by lead exposure, so as to provide the basis for the management of children's food safety.
Methods:
Based on the 2023 Dietary and Health Status Surveillance Project in Jiading District, children aged 3 to 4 years from 12 streets (townships) in Jiading District were selected as the subjects using a multi-stage stratified random sampling method. The consumption frequency and daily intake of seven subcategories of four major food groups (beans, tofu), cereals (wheat, rice), meat (animal viscera), and aquatic products (mantis shrimps, bivalves) consumed by children over the past three months were collected using a food frequency questionnaire. Food samples were collected according to the food safety risk monitoring plan in Jiading District, and the lead content was detected. The health risks of dietary lead exposure among 3-4-year-old children were quantitatively assessed using the single-factor pollution index method, the Nemerow comprehensive pollution index method, and the margin of safety (MOS) method.
Results:
A total of 143 3-4-year-old children were surveyed, including 69 boys (48.25%) and 74 girls (51.75%). A total of 317 food samples were tested, and lead was detected in all seven subcategories of food samples, with an overall detection rate of 77.29%. The detection rates of lead in bivalves and mantis shrimps of aquatic products were relatively high (98.75% and 100.00%, respectively). The mean lead content of various foods ranged from 0.003 4 to 0.090 7, with the highest level found in bivalves. The lead content of all food samples did not exceed the standard. The single-factor pollution index of seven subcategories ranged from 0.017 2 to 0.148 0, and the Nemerow comprehensive pollution index ranged from 0.116 5 to 0.424 4, both of which were less than 0.7. The MOS (mean) ranged from 0.000 3 to 0.003 9, with an overall MOS (mean) of 0.012 2. The MOS (P95) ranged from 0.005 7 to 0.055 9, with an overall MOS (P95) of 0.112 4, both of which were less than 1.
Conclusions
The lead pollution level in the diet among children aged 3 to 4 years in Jiading District is safe and clean, with an acceptable impact on food safety. However, the detection rate of lead is relatively high, and the main source of dietary lead exposure is aquatic products.
5.The sedative effect of remimazolam on ICU elderly patients undergoing mechanical ventilation and its influence on the circulatory system
Peng ZHAO ; Fangchao YAO ; Yi ZHENG ; Hailing DONG ; Jiuqing CUI ; Hao SUN ; Renjie LI ; Jingpu TIAN
Chinese Journal of Postgraduates of Medicine 2024;47(7):640-646
Objective:To investigate the sedative effect of remimazolam on ICU elderly patients undergoing mechanical ventilation and its influence on circulatory system.Methods:Using a prospective research approach, 189 ICU elderly patients undergoing mechanical ventilation in Hebei Petro China Central Hospital from October 2021 to June 2023 were selected. The patients were divided into remimazolam group, dexmedetomidine group and propofol group by random number table method with 63 cases in each group. The patients in remimazolam group, dexmedetomidine group and propofol group were sedated with remimazolam, dexmedetomidine and propofol, respectively. The sedation standard time, sedation standard rate, sedation maintenance time and recovery time after drug withdrawal were compared among the three groups. The heart rate, mean arterial pressure (MAP), respiratory rate and pulse oxygen saturation (SpO 2) before medication (T 0) and medication for 15 min (T 1), 30 min (T 2), 1 h (T 3), 6 h (T 4), 12 h (T 5) were recorded. The incidences of bradycardia, hypotension, respiratory depression, body movement and delirium during sedation were recorded. Results:The sedation standard time and recovery time after drug withdrawal in remimazolam group were significantly shorter than those in dexmedetomidine group and propofol group: (22.27 ± 5.31) min vs. (29.45 ± 6.24) and (30.12 ± 5.87) min, (28.66 ± 7.06) min vs. (32.22 ± 6.85) and (34.34 ± 7.24) min, and there were statistical differences ( P<0.05); there were no statistical difference between dexmedetomidine group and propofol group ( P>0.05). The sedation standard rate in remimazolam group and dexmedetomidine group was significantly higher than that in propofol group: 87.43% (661/756) and 83.60% (632/756) vs. 72.49% (548/756), and there was statistical difference ( P<0.016 7); there was no statistical difference between remimazolam group and dexmedetomidine group ( P>0.016 7). There was no statistical difference in sedation maintenance time among the three groups ( P>0.05). There were no statistical difference in T 0 heart rate, MAP, respiratory rate and SpO 2 among the three groups ( P>0.05). The T 1 to T 5 heart rate and MAP in remimazolam group were significantly higher than those in dexmedetomidine group and propofol group, the T 2 to T 5 heart rate and MAP in dexmedetomidine group were significantly lower than those in propofol group, and there were statistical differences ( P<0.05). The T 2 to T 5 respiratory rate in remimazolam group was significantly lower than that in dexmedetomidine group, the T 1 to T 5 respiratory rate in remimazolam group and dexmedetomidine group was significantly higher than that in propofol group, and there were statistical differences ( P<0.05). The T 2 to T 5 SpO 2 in remimazolam group and dexmedetomidine group was significantly higher than that in propofol group, and there was statistical difference ( P<0.05). The incidence of bradycardia in remimazolam group was significantly lower than that in dexmedetomidine group: 7.94% (5/63) vs. 25.40% (16/63), the incidence of hypotension was significantly lower than that in propofol group: 6.35% (4/63) vs. 23.81% (15/63), and there were statistical differences ( P<0.016 7). The incidence of respiratory depression in remimazolam group and dexmedetomidine group was significantly lower than that in propofol group: 4.76% (3/63) and 1.59% (1/63) vs. 22.22% (14/63), and there was statistical difference ( P<0.016 7). There was statistical difference in incidence of delirium among the three groups ( P<0.05), but there was no statistically significant difference in pairwise comparison ( P>0.016 7). There was no statistical difference in the incidence of body movement among the three groups ( P>0.05). Conclusions:The effect of remimazolam sedation in ICU elderly patients undergoing mechanical ventilation is satisfactory, with little influence on circulation and respiratory system and few adverse reactions.
6.Research on grading algorithm of diabetic retinopathy based on cross-layer bilinear pooling.
Liming LIANG ; Renjie PENG ; Jun FENG ; Jiang YIN
Journal of Biomedical Engineering 2022;39(5):928-936
Considering the small differences between different types in the diabetic retinopathy (DR) grading task, a retinopathy grading algorithm based on cross-layer bilinear pooling is proposed. Firstly, the input image is cropped according to the Hough circle transform (HCT), and then the image contrast is improved by the preprocessing method; then the squeeze excitation group residual network (SEResNeXt) is used as the backbone of the model, and a cross-layer bilinear pooling module is introduced for classification. Finally, a random puzzle generator is introduced in the training process for progressive training, and the center loss (CL) and focal loss (FL) methods are used to further improve the effect of the final classification. The quadratic weighted Kappa (QWK) is 90.84% in the Indian Diabetic Retinopathy Image Dataset (IDRiD), and the area under the receiver operating characteristic curve (AUC) in the Messidor-2 dataset (Messidor-2) is 88.54%. Experiments show that the algorithm proposed in this paper has a certain application value in the field of diabetic retina grading.
Humans
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Diabetic Retinopathy/diagnostic imaging*
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Algorithms
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ROC Curve
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Diabetes Mellitus
7.Electrospun gelatin polycaprolactone nanofiber aerogel combined with cartilage extracellular matrix for repair of cartilage injury in rabbits
Lei WANG ; Yi ZHANG ; Haining PENG ; Renjie CHEN ; Yunze LONG ; Zhong LIU ; Tengbo YU ; Yingze ZHANG
Chinese Journal of Trauma 2021;37(5):449-456
Objective:To investigate the repair effect of electrospun gelatin polycaprolactone (GT/PCL) nanofiber aerogels (NFA) combined with cartilage extracellular matrix (ECM) for treatment of cartilage injuries in rabbits.Methods:Firstly, the GT/PCL electrospun membrane was prepared by electrospinning and was ground into the short fiber at high speed. ECM was extracted and separated from fresh bovine articular cartilage, which mixed with the short fiber solution (10 ∶1). Subsequently, it was used to prepared GT/PCL/ECM (NFA) three-dimensional scaffold. Finally, the physical characteristics of the three different scaffolds (GT/PCL, ECM and GT/PCL/ECM) were detected by scanning electron microscope and Fourier-transform infrared (FTIR) spectrometer, including the composition, microstructure, swelling rate, porosity, compressive strength and degradation rates. And the biocompatibility research was getting on by co-culturing the scaffold with chondrocytes. Fifteen male New Zealand white rabbits were divided into blank control group (Group A, n=5), ECM group (group B, n=5) and composite scaffold(GT/PCL/ECM)group (Group C, n=5) according to the random number table. An injury model was established and three types of bio-scaffold materials were implanted into different groups. At 3 weeks, the cartilage repair was evaluated among groups by semi quantitative global MRI scoring system (WORMS). After the animals were killed, the knee joints of each group were scored by the international society for cartilage repair histological score (ICRs); the ICRs histological score was performedby HE staining and safranine green staining. Results:Three scaffolds showed a porous three-dimensional structure under the scanning electron microscope. FIRT showed that GT and PCL were introduced into the scaffolds successfully. The GT/PCL NAF was loose and unable to be characterized by materials science. The swelling rate of GT/PCL/ECM scaffold [(1, 092.0±32.2)%] was higher than that of ECM scaffold [(933.6±16.3)%] ( P<0.01). The porosity of GT/PCL/ECM scaffold [(92.3±2.3)%] was higher than that of ECM scaffold [(85.9±2.2)%] ( P<0.05). The compressive strength of ECM scaffold [(2.7±0.1)kPa] and of GT/PCL/ECM scaffold [(2.4±0.1)kPa] showed no statistical difference ( P>0.05). The degradation rate of ECM scaffold was higher than that of GT/PCL/ECM scaffold, but the difference was not statistically significant ( P>0.05). The cytotoxicity rating of GT/PCL/ECM scaffold was grade I, indicating that its biocompatibility was better. At 3 weeks, the MRI WORMS score in Group C [(49.0±11.4)points] was significantly higher than that in Group B [(40.0±6.7)points] and that in Group A [(24.0±6.5) points] ( P<0.05 or 0.01); the general ICRS score of group C was [(7.4±1.1) points], which was significantly higher than that of group B [(4.6±1.1)points] and group A [(3.0±1.2)points] ( P<0.01); The ICRS histological scores of group C and group B were [(6.8±0.8)points] and [(4.2±0.8)points] respectively compared with group A [(2.8±0.8)points] were significantly higher ( P<0.05 or 0.01). Conclusion:GT/PCL/ECM (NFA) scaffold has similar tissue structure to natural cartilage and is superior to traditional ECM scaffold in physical properties and biocompatibility, which provides a stable environment for chondrocyte adhesion and growth, promotes collagen regeneration, and thus accelerates the repair of cartilage injury.
8.Diffusion tensor imaging and resting-state functional magnetic resonance imaging in patients with delirium in intensive care unit
Renjie SONG ; Ganjun SONG ; Peng XIE ; Haizhen DUAN ; Tianxi ZHANG ; Yuanlan LU ; Mo LI ; Xiaoyun FU
Chinese Critical Care Medicine 2020;32(1):88-93
Objective:To analyze the brain function of patients with delirium in intensive care unit (ICU) using resting-state functional magnetic resonance imaging (fMRI), further analyze the structural changes in the brain using diffusion tensor imaging (DTI), and explore the correlations of brain function with structural changes in patients with delirium in ICU from a new perspective of functional imaging, provide visual evidence for the diagnosis of delirium.Methods:Patients with delirium admitted to ICU of the Affiliated Hospital of Zunyi Medical University from January 1st to December 31st in 2017 were enrolled as subjects. During the same period, the healthy volunteers who matched the gender, age and education level of the patients with delirium were enrolled as control group. The intensive care delirium screening checklist (ICDSC) scores within 24 hours after ICU admission were recorded. All the subjects were scanned by fMRI and DTI. The abnormal changes in resting-state brain function of the patients with delirium were evaluated by cerebral regional homogeneity (ReHo) data analysis. The DTI data were processed by the FSL software, and the fractional anisotropy (FA) and mean diffusivity (MD) of the brain were extracted, respectively, to evaluate the damage to brain structure. The values of ReHo, FA and MD were compared between the two groups. The ReHo value of brain region with reduced ReHo value of patients with delirium as compared with the healthy volunteers was extracted for Pearson correlation analysis with ICDSC scores.Results:A total of 22 patients with delirium were included. Seven patients who did not cooperate in the examination, used sedatives or had false images in scanning, were excluded. Finally, 15 patients were enrolled in the delirium group, and 15 healthy volunteers in the healthy control group. ① No statistically significant difference was found in gender, age or education time between the two groups. ICDSC score of the delirium group was significantly higher than that of the healthy control group (6.07±1.28 vs. 1.07±0.88, P < 0.01). ② fMRI scanning and analysis results: compared with the healthy control group, the ReHo values of the cerebellum, right hippocampus, striatum, midbrain and pons in the delirium group were significantly increased (all P < 0.05, AlphaSim correction), while the ReHo values of bilateral superior frontal gyrus, bilateral median frontal gyrus, left inferior frontal gyrus, temporal lobe and parietal lobe were significantly lowered (all P < 0.05, AlphaSim correction). Correlation analysis showed that the ReHo value of the left superior frontal gyrus was negatively correlated with ICDSC score in the patients with delirium ( r = -0.794, P < 0.05), indicating that the changes in the functional area of the medial frontal gyrus was most closely related to delirium. ③ DTI scanning and analysis results: compared with the healthy control group, the FA values of the left cerebellum, bilateral frontal lobes, left temporal lobe, corpus callosum and left hippocampus in the delirium group were decreased significantly (all P < 0.05, AlphaSim correction), while the MD values of the medial frontal gyrus, right superior temporal gyrus, anterior cingulate gyrus, bilateral insular lobes and left caudate nucleus were enhanced significantly (all P < 0.05, AlphaSim correction), suggesting that the structural and functional damage was found in multiple brain regions in patients with delirium. Conclusions:Multiple brain regions of patients with delirium present abnormal resting-state brain function. The abnormal resting-state brain function of the left superior frontal gyrus is closely related to the occurrence of delirium. Structural damage is found in multiple brain regions of patients with delirium. The structural changes in the frontal lobe, temporal lobe, corpus callosum, hippocampus and cerebellum and their abnormal functions can be used as preliminary imaging indexes for the diagnosis of delirium.
9.Experimental study on the protective effect of EPO in glucocorticoid-induced osteoporosis
Yongqing YAN ; Renjie XU ; Peng WEI
Chinese Journal of Orthopaedics 2020;40(21):1469-1477
Objective:To study the mechanism of protective effect of erythropoietin (EPO) on glucocorticoid-induced osteoporosis in rats through promoting vascular action mediated by VEGF, promoting osteogenesis, and inhibiting bone resorption.Methods:An animal experiment of 24 SD rats in total were divided into three groups: the osteoporosis group (20 mg/kg BW of methylprednisolone biwim for 6 weeks); the EPO group (MP 20 mg/kg biw+EPO 500 u/kg qdim); the nature salt group (0.9%NS). After 12 weeks, rats were harvested and received examination of histology (HE staining) for demonstration of protective effect, immunohistochemistry with CD31 stainingfor microvascular changes ,changes of VEGF and PCNA expressions using Western Boltfor microvascular and cell proliferation, and ELISA to detectOPN, PINP, CTX-1 in serumasbone turnover marker.Results:Hematoxylin and eosin staining in the model group showed that the bony trabeculae had become obviously narrow and sparse with discontinuity of the integrity. The integrity of the trabeculae was better in the EPO group. Immunohistochemical results: the EPO group CD31+ cell 16.60±4.88, the osteoporosis group 12.96±4.54, the NS group 25.84±7.97. CD31 expression was higher in the EPO group than the osteoporosis group. Western Bolt: in the NS groupthe ratio of VEGF/β-actin greyscalewas 0.570±0.022, with the osteoporosis group 0.446±0.083 and the EPO group 0.584±0.009; The ratio of PCNA/β-actingreyscale was 0.541±0.158 in the NS group, withthe osteoporosis group 0.187±0.099, the EPOgroup 0.733±0.257. VEGF and PCNA expression in the EPO group were higher than those in the osteoporosis group. ELISA: OPN results: the NS group 78.34±17.28 pg/ml, the osteoporosis group 368.48±97.23 pg/ml, the EPO group 217.62±39.11 pg/ml; P1NP results: the NS group 1 507.00±58.49 ng/ml, the osteoporosis group 1 196.00±91.32 ng/ml, the EPO group 1 621.00±65.57 ng/ml; CTX-1 results: the NS group 27.10±4.78 ng/ml, the osteoporosis group 39.46±9.23 ng/ml, the EPO group 31.17±4.11 ng/ml. The level of OPN and CTX-1 in the EPO group were lower than that in the osteoporosis group, and P1NP was higher than that in the osteoporosis group.Conclusion:EPO generates certain protective effect on bone of rats withglucocorticoid-induced osteoporosis. Its potential mechanism is to promote vascular action, promote osteogenesis, and inhibit bone resorption.
10.LINC00703 Acts as a Tumor Suppressor via Regulating miR-181a/KLF6 Axis in Gastric Cancer
Haiyang YANG ; Minqi PENG ; Yanjiao LI ; Renjie ZHU ; Xiang LI ; Zhengjiang QIAN
Journal of Gastric Cancer 2019;19(4):460-472
PURPOSE: Long noncoding RNA 00703 (LINC00703) was found originating from a region downstream of Kruppel-like factor 6 (KLF6) gene, having 2 binding sites for miR-181a. Since KLF6 has been reported as a target of miR-181a in gastric cancer (GC), this study aims to investigate whether LINC00703 regulates the miR-181a/KLF6 axis and plays a functional role in GC pathogenesis.MATERIALS AND METHODS: GC tissues, cell lines, and nude mice were included in this study. RNA binding protein immunoprecipitation (RIP) and pull-down assays were used to evaluate interaction between LINC00703 and miR-181a. Quantitative real-time polymerase chain reaction and western blot were applied for analysis of gene expression at the transcriptional and protein levels. A nude xenograft mouse model was used to determine LINC00703 function in vivo.RESULTS: We revealed that LINC00703 competitively interacts with miR-181a to regulate KLF6. Overexpression of LINC00703 inhibited cell proliferation, migration/invasion, but promoted apoptosis in vitro, and arrested tumor growth in vivo. LINC00703 expression was found to be decreased in GC tissues, which was positively correlated with KLF6, but negatively with the miR-181a levels.CONCLUSIONS: LINC00703 may have an anti-cancer function via modulation of the miR-181a/KLF6 axis. This study also provides a new potential diagnostic marker and therapeutic target for GC treatment.
Animals
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Apoptosis
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Binding Sites
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Blotting, Western
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Cell Line
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Cell Proliferation
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Gene Expression
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Heterografts
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Immunoprecipitation
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In Vitro Techniques
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Mice
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Mice, Nude
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Real-Time Polymerase Chain Reaction
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RNA, Long Noncoding
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RNA-Binding Proteins
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Stomach Neoplasms


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