Objective:The neurovascular unit(NVU)damage in rats with cerebral ischemia-reperfusion injury(CI-RI)was evaluated based on multimodal MRI combined with multiple histological techniques to provide visual evidence for clinical assessment of CIRI.Methods:28 healthy male Sprague-Dawley rats were randomly divided into sham-oper-ated(Sham)group and CIRI group.At 24 h after modeling,rats underwent head T2-weighted imaging,diffusion-weighted imaging,water extraction with phase-contrast arterial spin labeling magnetic resonance imaging(MRI),and cerebral blood bleeding oxygen monitoring.Hematoxylin-eosin(HE)staining was used to observe the morphological changes of nerve cells in cerebral cortex.Immunofluorescence staining and transmission electron microscopy were used to observe the changes of NVU after CIRI.The endothelial tight junction proteins(claudin-5 and ZO-1)and matrix metalloproteinase-9(MMP-9)were detected by immunohistochemical staining and Western Blot analysis,to evaluate blood-brain-barrier(BBB)disruption in CIRI rats.Results:The MRI results showed that the brain tissue of the CIRI group was edematous,and the blood perfusion significantly decreased and the BBB permeability significantly increased compared with that of the Sham group(P＜0.01).The results of cerebral blood hemoglobin oxygen monitoring showed that the hemoglobin oxygen saturation of the cerebral cortex on the injured side was significantly lower than that of the Sham group(P＜0.05).HE staining showed the neuronal cells of the injured cerebral cortex were disordered and the nuclei shrank.Immunofluorescence staining results showed the number of NeuN and CD31 positive cells in the cerebral cortex of the injured side was significantly lower than that in the Sham group(P＜0.01),and the number of TUNEL positive cells in the cerebral cortex and striatum was significantly higher than that in the Sham group(P＜0.05).Transmission electron microscopy revealed the tight junctions of cerebral cortical endothelial cells were damaged,neuro-nal nucleus contraction,and astrocyte body edema.Immunohistochemical staining showed that the number of claudin-5 and ZO-1 positive cells in the cerebral cortex on the injured side was significantly lower than that in the Sham group,and the expression of MMP-9 was significantly higher than that in the Sham group(P＜0.01).Conclusion:CIRI re-sults in NVU injury,BBB disruption,and exacerbation of brain damage in rats.Multimodal MRI combined with various histopathological techniques effectively assesses NVU injury after CIRI.

Objective:The neurovascular unit(NVU)damage in rats with cerebral ischemia-reperfusion injury(CI-RI)was evaluated based on multimodal MRI combined with multiple histological techniques to provide visual evidence for clinical assessment of CIRI.Methods:28 healthy male Sprague-Dawley rats were randomly divided into sham-oper-ated(Sham)group and CIRI group.At 24 h after modeling,rats underwent head T2-weighted imaging,diffusion-weighted imaging,water extraction with phase-contrast arterial spin labeling magnetic resonance imaging(MRI),and cerebral blood bleeding oxygen monitoring.Hematoxylin-eosin(HE)staining was used to observe the morphological changes of nerve cells in cerebral cortex.Immunofluorescence staining and transmission electron microscopy were used to observe the changes of NVU after CIRI.The endothelial tight junction proteins(claudin-5 and ZO-1)and matrix metalloproteinase-9(MMP-9)were detected by immunohistochemical staining and Western Blot analysis,to evaluate blood-brain-barrier(BBB)disruption in CIRI rats.Results:The MRI results showed that the brain tissue of the CIRI group was edematous,and the blood perfusion significantly decreased and the BBB permeability significantly increased compared with that of the Sham group(P＜0.01).The results of cerebral blood hemoglobin oxygen monitoring showed that the hemoglobin oxygen saturation of the cerebral cortex on the injured side was significantly lower than that of the Sham group(P＜0.05).HE staining showed the neuronal cells of the injured cerebral cortex were disordered and the nuclei shrank.Immunofluorescence staining results showed the number of NeuN and CD31 positive cells in the cerebral cortex of the injured side was significantly lower than that in the Sham group(P＜0.01),and the number of TUNEL positive cells in the cerebral cortex and striatum was significantly higher than that in the Sham group(P＜0.05).Transmission electron microscopy revealed the tight junctions of cerebral cortical endothelial cells were damaged,neuro-nal nucleus contraction,and astrocyte body edema.Immunohistochemical staining showed that the number of claudin-5 and ZO-1 positive cells in the cerebral cortex on the injured side was significantly lower than that in the Sham group,and the expression of MMP-9 was significantly higher than that in the Sham group(P＜0.01).Conclusion:CIRI re-sults in NVU injury,BBB disruption,and exacerbation of brain damage in rats.Multimodal MRI combined with various histopathological techniques effectively assesses NVU injury after CIRI.

Humans ; Female ; Breast Neoplasms/pathology* ; Multiomics ; Trastuzumab ; Receptor, ErbB-2/genetics* ; Machine Learning

Objective:To compare the effects of Feishu(BL13)combined with Tianshu(ST25)and Feishu(BL13)alone on pulmonary function and tissue inflammation in asthma model rats. Methods:Forty-eight Sprague-Dawley rats were randomly divided into a normal group,a model group,a lung treatment group,and a lung-intestine treatment group according to the random number table method,with 12 rats in each group.Except for the normal group,rats in the other three groups were sensitized by ovalbumin followed by atomization stimulation to establish the asthma model.After the model was made successfully,rats in the lung treatment group were treated with acupuncture at bilateral Feishu(BL13)for 30 min;rats in the lung-intestine treatment group were treated with acupuncture at bilateral Feishu(BL13)and Tianshu(ST25)for 15 min,respectively,for 30 min in total.Acupuncture was performed once a day for 14 d.Rats in the other two groups did not receive any intervention.After intervention,the pulmonary function of each group of rats was measured by a pulmonary function tester.The histopathological changes stained by hematoxylin-eosin staining and the collagen deposition degree stained by Masson in lung tissue,as well as the inflammatory cells in the bronchoalveolar lavage fluid(BALF)stained by Wright's-Giemsa staining were observed under a light microscope;the levels of interleukin(IL)-4,IL-5,IL-13,IL-17,IL-25,IL-33,leukotrienes(LT),thymic stromal lymphopoietin(TSLP),and prostaglandin D2(PGD2)were detected by enzyme-linked immunosorbent assay. Results:Compared with the normal group,the peak expiratory flow(PEF),the dynamic lung compliance(Cdyn),the forced expiratory flow 25%(FEF25%),the forced expiratory volume in the first second/forced vital capacity(FEV1/FVC),and the maximum mid-expiratory flow(MMEF)in the model group were significantly decreased(P<0.01 or P<0.05);the lung resistance(RL),the collagen deposition,the IL-4,IL-5,IL-13,IL-17,IL-33,LT,TSLP,and PGD2 levels,and the neutrophil proportion in the BALF were increased significantly(P<0.01 or P<0.05).Compared with the model group,the FEF25%and FEV1/FVC in the pulmonary function were significantly increased(P<0.01,P<0.05),while the collagen fiber deposition and the levels of IL-4,IL-5,IL-13,IL-17,LT,TSLP,and PGD2 were significantly decreased(P<0.01 or P<0.05)in the lung treatment group;the PEF,FEF25%,and FEV1/FVC in pulmonary function were significantly increased(P<0.01 or P<0.05),while the RL and the collagen fiber deposition,as well as the neutrophil proportion and the levels of IL-4,IL-5,IL-13,IL-17,IL-33,LT,TSLP,and PGD2 in the BALF were significantly decreased(P<0.01 or P<0.05)in the lung-intestine treatment group.Compared with the lung treatment group,the IL-5 level in the rat lung tissue of the lung-intestine treatment group was decreased(P<0.05). Conclusion:Acupuncture in the lung treatment and the lung-intestine treatment groups improved pulmonary function,lung inflammation,and inflammatory cells in the BALF in asthma model rats;the lung-intestine treatment was better than lung treatment in improving the IL-5 level.The combination of Feishu(BL13)and Tianshu(ST25)was superior to Feishu(BL13)alone in the treatment of asthma in trend.

Objective: To evaluate the impact of bleomycin/etoposide/cisplatin (BEP) and paclitaxel/carboplatin (PC) chemotherapy regimens on the fertility and prognostic outcomes in malignant ovarian germ cell tumor (MOGCT) patients who underwent fertility-sparing surgery (FSS). Methods: A propensity score matching algorithm was performed between the BEP and PC groups. The χ2 test and the Kaplan-Meier method were used to compare the fertility outcome, disease-free survival (DFS) and overall survival (OS). The Cox proportional hazards regression analysis was used to identify risk factor of DFS. Results: We included 213 patients, 185 (86.9%) underwent BEP chemotherapy, and 28 (13.1%) underwent PC chemotherapy. The median age was 22 years (range, 8–44 years), and the median follow-up period was 63 months (range, 2–191 months). Fifty-one (29.3%) patients had a pregnancy plan, and 35 (85.4%) delivered successfully. In the before and after propensity score matching cohorts, there were no significant differences in spontaneous abortion, selective termination of pregnancy, during-pregnancy status, and live birth between the BEP and PC groups (p>0.05). Fourteen (6.6%) patients experienced recurrence, including 11 (5.9%) in the BEP group and 3 (10.7%) in the PC group. Four (1.9%) patients in the BEP group died. Kaplan-Meier analysis revealed no significant differences in DFS (p=0.328) and OS (p=0.446) between the BEP and PC groups, and the same survival results were observed in the after matching cohort. Conclusion The PC regimen is as safe as the BEP regimen for MOGCT patients with fertility preservation treatment, and no differences were observed in fertility and clinical prognosis.

Objective:To understand the molecular epidemic characteristics and genes of human astrovirus (HAstV) in infantile diarrhea patients in Jilin province from 2017 to 2020.Methods:Fecal samples of hospitalized infants with diarrhea under 5 years of age from January 2017 to December 2020 were collected. The nucleic acids of fecal samples were amplified by reverse transcription-polymerase chain reaction (RT-PCR), the positive amplification products were sequenced, and the phylogenetic tree was constructed.Results:A total of 2 019 fecal samples were collected, of which 71 were positive for HAstV, with a positive rate of 3.5%. Among the cases with positive samples of HAstV, 42 had mixed infection with other viruses causing diarrhea, accounting for 59.2% of the total number of positive cases, half of which were mixed infection with rotavirus and HAstV. From 2017 to 2020, the positive rates of HAstV were 4.69%, 1.98%, 5.93% and 0.43% per year, respectively. There were two epidemic peaks every year and one epidemic peak year every two years. The positive rate of 36-47 months old was the highest, followed by 0-2 months old. A total of 55 sequences were obtained in this study. Phylogenetic tree analysis showed that they all belonged to classical HAstV, including 42 HAstV-1a subtypes, 8 HAstV-1b subtypes and 5 HAstV-5 subtypes.Conclusions:HAstV is one of the important pathogens of diarrhea in children under 5 years of age in Jilin province. The epidemic law has obvious seasonal and age differences. The dominant strain is HAstV-1a.

Objective:To summarize and analyze the clinical data of Chinese patients with colony-stimulating factor 1 receptor (CSF1R)-related leukoencephalopathy, and clarify the phenotypic and genetic characteristics of Chinese patients.Methods:Medical history of patients with CSF1R-related leukoencephalopathy diagnosed from April 1, 2018 to January 31, 2021 in the department of neurology of 22 hospitals in China was collected, and scores of Mini-Mental State Examination (MMSE), Montreal Cognitive Assessment Scale (MoCA), magnetic resonance severity scale were evaluated. Group comparison was performed between male and female patients.Results:A total of 62 patients were included, and the male-female ratio was 1∶1.95. The age of onset was (40.35±8.42) years. Cognitive impairment (82.3%, 51/62) and motor symptoms (77.4%,48/62) were the most common symptoms. The MMSE and MoCA scores were 18.79±7.16 and 13.96±7.23, respectively, and the scores of two scales in male patients (22.06±5.31 and 18.08±5.60) were significantly higher than those in females (15.53±7.41 , t=2.954, P=0.006; 10.15±6.26, t=3.328 , P=0.003). The most common radiographic feature was bilateral asymmetric white matter changes (100.0%), and the magnetic resonance imaging severity scale score was 27.42±11.40, while the white matter lesion score of females (22.94±8.39) was significantly higher than that of males (17.62±8.74 , t=-2.221, P<0.05). A total of 36 CSF1R gene mutations were found in this study, among which c.2381T>C/p.I794T was the hotspot mutation that carried by 17.9% (10/56) of the probands. Conclusions:The core phenotypic characteristics of CSF1R-related leukoencephalopathy in China are progressive motor and cognitive impairment, with bilateral asymmetrical white matter changes. In addition, there exist gender differences clinically, with severer cognitive impairment and imaging changes in female patients. Thirty-six CSF1R gene mutations were found in this study, and c.2381T>C/p. I794T was the hotspot mutation.

Objective:To study the biological behavior of nasopharyngeal carcinoma stem cells and to explore the activation of Ras signaling pathway regulated by CD44.Methods:CNE2-SC and 5-8F-SC were nasopharyngeal carcinoma stem cells and obtained by serum-free suspension culture. Cell counting kit-8 (CCK-8) assay, colony formation assay, Transwell migration assay, cell adhesion array were used to investigate the growth, proliferation, migration and adhesion of nasopharyngeal carcinoma stem cells. Western blot test was used to detect the expressions of Ras signaling pathway related proteins and siRNA-mediated interference was used to determine the activation of Ras signaling pathway regulated by CD44.Results:The growth rates of CNE2-SC and 5-8F-SC cells were significantly lower than those of nasopharyngeal carcinoma cells at 24, 48 and 72 hours after inoculation ( P<0.05). After 14 days of implantation, the colony formation rates of CNE2-SC (44.5±1.9)% and 5-8F-SC (47.4±1.8)% were higher than those of CNE2 (34.9±1.5)% and 5-8F (37.2±1.7)%, respectively( P<0.01). The migration cell number of CNE2-SC was (87.6±7.8), 3.97 times higher than that of CNE2 ( P<0.01). The migration cell number of 5-8F-SC was (67.2±5.7), 3.07 times higher than 5-8F ( P<0.01). The adhesion rates of CNE2-SC and CNE2 cells were (42.1±7.6)% and (8.9±2.0)%, respectively at 3 hours after inoculation and were (82.4±5.0)% and (12.1±2.2)% at 6 hours after inoculation, respectively. The adhesion rate of CNE2-SC cells was higher than that of CNE2 cells (all P<0.01). The adhesion rates of 5-8F-SC and 5-8F cells were (53.6±6.1)% and (7.3±1.5)% at 3 hours after inoculation, and (90.7±3.6)% and (11.0±1.2)% at 6 hours after inoculation, respectively. The adhesion rate of 5-8F-SC cells was higher than that of 5-8F cells ( P<0.01). The expression levels of CD44, Ras and N-cadherin were significantly higher, while phosphatase and tensin homolog deleted on chromosome 10 (PTEN), E-cadherin in nasopharyngeal carcinoma stem cells were lower than those of the nasopharyngeal carcinoma cells. Furthermore, the levels of phosphorylated mitogen extracellular kinase1/2 (p-MEK1/2) and phosphorylated extracellular signal-regulated protein kinase1/2 (p-ERK1/2)were significantly increased in nasopharyngeal carcinoma stem cells ( P<0.01). Correlation analysis showed that the protein expression levels of CD44 was highly positively correlated with RAS in nasopharyngeal carcinoma stem cells( r=0.985, P=0.002; r=0.962, P=0.038). Deletion of CD44 in CNE2-SC decreased the expression levels of HER-2, Ras and p-ERK1/2, p-Akt and phosphorylated protein kinase C-δ(p-PKCδ) ( P<0.01). Conclusions:Despite compare to the nasopharyngeal carcinoma cell, nasopharyngeal carcinoma stem cells grows at a relatively slow rate, the capacities of clone formation, migration, adhesion are promoted. This may be related to the CD44-regulated abnormal activation of Ras signaling pathway.

Objective:To study the biological behavior of nasopharyngeal carcinoma stem cells and to explore the activation of Ras signaling pathway regulated by CD44.Methods:CNE2-SC and 5-8F-SC were nasopharyngeal carcinoma stem cells and obtained by serum-free suspension culture. Cell counting kit-8 (CCK-8) assay, colony formation assay, Transwell migration assay, cell adhesion array were used to investigate the growth, proliferation, migration and adhesion of nasopharyngeal carcinoma stem cells. Western blot test was used to detect the expressions of Ras signaling pathway related proteins and siRNA-mediated interference was used to determine the activation of Ras signaling pathway regulated by CD44.Results:The growth rates of CNE2-SC and 5-8F-SC cells were significantly lower than those of nasopharyngeal carcinoma cells at 24, 48 and 72 hours after inoculation ( P<0.05). After 14 days of implantation, the colony formation rates of CNE2-SC (44.5±1.9)% and 5-8F-SC (47.4±1.8)% were higher than those of CNE2 (34.9±1.5)% and 5-8F (37.2±1.7)%, respectively( P<0.01). The migration cell number of CNE2-SC was (87.6±7.8), 3.97 times higher than that of CNE2 ( P<0.01). The migration cell number of 5-8F-SC was (67.2±5.7), 3.07 times higher than 5-8F ( P<0.01). The adhesion rates of CNE2-SC and CNE2 cells were (42.1±7.6)% and (8.9±2.0)%, respectively at 3 hours after inoculation and were (82.4±5.0)% and (12.1±2.2)% at 6 hours after inoculation, respectively. The adhesion rate of CNE2-SC cells was higher than that of CNE2 cells (all P<0.01). The adhesion rates of 5-8F-SC and 5-8F cells were (53.6±6.1)% and (7.3±1.5)% at 3 hours after inoculation, and (90.7±3.6)% and (11.0±1.2)% at 6 hours after inoculation, respectively. The adhesion rate of 5-8F-SC cells was higher than that of 5-8F cells ( P<0.01). The expression levels of CD44, Ras and N-cadherin were significantly higher, while phosphatase and tensin homolog deleted on chromosome 10 (PTEN), E-cadherin in nasopharyngeal carcinoma stem cells were lower than those of the nasopharyngeal carcinoma cells. Furthermore, the levels of phosphorylated mitogen extracellular kinase1/2 (p-MEK1/2) and phosphorylated extracellular signal-regulated protein kinase1/2 (p-ERK1/2)were significantly increased in nasopharyngeal carcinoma stem cells ( P<0.01). Correlation analysis showed that the protein expression levels of CD44 was highly positively correlated with RAS in nasopharyngeal carcinoma stem cells( r=0.985, P=0.002; r=0.962, P=0.038). Deletion of CD44 in CNE2-SC decreased the expression levels of HER-2, Ras and p-ERK1/2, p-Akt and phosphorylated protein kinase C-δ(p-PKCδ) ( P<0.01). Conclusions:Despite compare to the nasopharyngeal carcinoma cell, nasopharyngeal carcinoma stem cells grows at a relatively slow rate, the capacities of clone formation, migration, adhesion are promoted. This may be related to the CD44-regulated abnormal activation of Ras signaling pathway.

Objective To investigate the correlation between cilioretinal artery and central visual impairment in patients with central retinal artery occlusion (CRAO).Methods A total of 32 CRAO patients (32 eyes) were selected,and results of central vision and fundus fluorescein angiography (FFA) were analyzed retrospectively.Correlation between cilioretinal arteries and degree of central visual acuity impairment was analyzed.Results In the 32 CRAO eyes,11 eyes were found with cilioretinal? arteries.Eye examination results showed that there were 10 eyes with mild visual impairment,9 eyes with moderate visual impairment and 13 eyes with severe visual impairment in the total 32 eyes.There were significant difference in the degrees of central visual impairment between CRAO patients with or without cilioretinal artery (P ＜ 0.05),and there was a significant correlation between cilioretinal artery and degree of central visual acuity impairment.Conclusion 34.4％ CRAO patients have cilioretinal artery,and cilioretinal artery in CRAO patients is associated with the degree of central visual acuity impairment.