[Contribution of central motion conduction time to the assessment of corticospinal tract lesions and its clinical significance].

Objective: To investigate the association of central motion conduction time (CMCT) with corticospinal tract lesions and its clinical application. Methods: Patients who completed transcranial magnetic stimulation-motor evoked potentials were included from Department of Neurology, Xuanwu Hospital between June 2020 and June 2021. The differences of CMCT values between corticospinal tract sign-positive group and tendon reflex-positive group and the relevant negative groups were compared. The consistency between increased CMCT values and the positive signs of corticospinal tract damage, as well as the significance of CMCT in different neurological diseases were further evaluated. Results: A total of 271 patients were included in the study, aged 12-86 (49±16) years, with 137 males (50.55%) and 134 females (49.45%). The CMCT valuesï¼»M（Q1，Q3）］from Hoffmann's sign-positive group [9.52 (8.54, 10.99) ms vs 9.03 (8.30, 9.53) ms], Babinski's sign-positive group [19.54 (16.97, 24.43) ms vs 16.85(15.63, 18.55) ms] and tendon reflex-positive group [15.38 (9.27, 19.28) ms vs 10.49(8.79, 16.60) ms] were larger than those of relevant negative groups (all P<0.01). In the Babinski sign-positive group, 78.01%(181/232) of the patients had increased CMCT, while in the Hoffmann's sign-positive group, only 26.03%(19/73) of the patients had increased CMCT, indicating that the contribution of CMCT from the lower extremities to the assessment of corticospinal tract lesions was better than that of the upper extremities. With the increase of CMCT values in lower limbs, Babinski sign positive rate increased, the difference was statistically significant(P<0.001). In nervous system diseases, the consistency between CMCT and pathological signs was 75.65% (205/271). Conclusions: The contribution of CMCT from the lower extremities to the assessment of corticospinal tract lesions is superior to that of upper limbs. The higher increase of CMCT values are more reliable for corticospinal tract damage. CMCT has a good concordance with corticospinal tract lesions in some neurological diseases, which can be used to assist clinical diagnosis.

Tunable quad-band transmission response, based on single-layer metamaterials.

We investigated the electromagnetically induced transparency (EIT)-like effects in planar metamaterials (MMs) at microwave (GHz) frequencies. The specific MMs that were used in this study consist of cut-wire resonator/ring resonator, which achieved the dual EIT-like effects in a single-layer through the bright- and quasi-dark-mode coupling and the lattice mode coupling. In addition, by varying the distance between the two resonators, the quad-band EIT spectral response in the microwave region was obtained, and the group refractive index at the EIT-like resonance of proposed design reached up to 4,000. This study provides the design approach to the multispectral EIT-like effects and might suggest potential applications in a variety of fields, for example, low-loss slow-light device, multiple switching sensor, and other sensing devices.

Production of medically useful nitric monoxide using AC arc discharge.

Inhaled nitric monoxide (iNO) is increasingly used as a medical treatment for acute respiratory distress syndrome. A course of the existing nitric monoxide (NO) therapy with gas cylinders could cost up to approximately $15,000 for an average of 30.2 h. Moreover, a gas cylinder containing a mixture of N2 and NO may potentially leak NO. The objective of this study is to develop an efficient and cost-effective on-site iNO generation system. In the present setup, NO was generated by using dry air or mixed oxygen/nitrogen (O2/N2) and an AC power source with an output power level of 5-30 W at atmospheric pressure. The simultaneously produced NO2 was eliminated with an ammonium sulfite ((NH4)2SO3) solution. The effects of the O2/N2 ratio, gas flow rate, discharge gap distance, output energy density and electrode structure on NOx concentration and the NO/NO2 ratio are reported. The concentrations of NO and NO2 reached 62 ppm and 3 ppm, respectively, after absorption and dilution at a gas flow rate of 6 L/min. With the present setup, the AC arc discharge produced NOx at a stable concentration for at least 6 h using dry air.

Efficacy of 3D VIBE Dixon fat quantification for differentiating clear-cell from non-clear-cell renal cell carcinoma.

AIM: To assess the efficacy of three-dimensional (3D) volumetric interpolated breath-hold examination (VIBE) magnetic resonance imaging (MRI) with Dixon quantification for differentiating clear-cell from non-clear-cell types of renal cell carcinoma (RCC). MATERIALS AND METHODS: The 3D VIBE Dixon renal MRI examinations of 44 patients with 45 histologically confirmed RCCs was analysed. The fat fractions and signal intensity indexes (SIindex) of the solid portions of clear-cell and non-clear-cell RCCs were measured and compared using Student's t-test and receiver operating characteristic (ROC) curves. The agreement of measurements among observers was evaluated by the intraclass correlation coefficient (ICC), and Bland-Altman plots. RESULTS: The mean values of fat fraction (13.16±7.16%) and SIindex (22.64±15.7%) in clear-cell RCCs were significantly higher than that in non-clear-cell RCCs (7.7±2% and 7.9±4.8%; p<0.001, respectively). With the area under the ROC curve (AUC) of the fat fraction at 0.811, 75% (95% CI: 55.1-89.43%) sensitivity and 76.5% (95% CI: 50.1-93.2%) specificity for diagnosing clear-cell RCC were obtained at a cut-off fat fraction value of 8.9%. With a cut-off value of 8.89%, the diagnostic sensitivity and specificity were 85.7% (95% CI: 67.3-96%) and 70.6% (95% CI: 44-89.7%), respectively. The AUC of the SIindex was 0.870 (0.766-0.973). ICC and Bland-Altman plots show excellent agreement of the tumour fat fraction and SIindex measurement between the two observers. CONCLUSION: Intracellular lipid content analysis using the 3D Dixon technique can help to differentiate clear-cell from non-clear-cell RCCs.

Preliminary study of diffusion-weighted MRI in the preoperative diagnosis of cystitis glandularis.

AIM: To assess the efficacy of diffusion-weighted (DWI) magnetic resonance imaging (MRI) in distinguishing cystitis glandularis (CG) from bladder urothelial carcinoma. MATERIALS AND METHODS: Ultrasound, computed tomography (CT), conventional MRI, and DWI of 30 patients with histopathologically confirmed CG were analysed retrospectively and the imaging findings were correlated to the findings at histology. RESULTS: Ultrasound was non diagnostic in 11/18 and misdiagnosed malignancy in 7/18; CT was non diagnostic in 6/10 and misdiagnosed malignancy in 4/10; MRI was non diagnostic in 0 and misdiagnosed malignancy in 4/5 respectively. One patient with diffuse bladder wall thickening was correctly diagnosed as CG at MRI. All six patients who underwent additional DWI were accurately diagnosed as having CG with no or minimal reduction of diffusion. CONCLUSIONS: Diffusion is not reduced or shows minimal reduction in CG. DWI may aid the differential diagnosis of CG.

[Endoplasmic reticulum stress in INS-1-3 cell associated with the expression changes of MODY gene pathway].

Objective: To profile the gene expression changes associated with endoplasmic reticulum stress in INS-1-3 cells induced by thapsigargin (TG) and tunicamycin (TM). Methods: Normal cultured INS-1-3 cells were used as a control. TG and TM were used to induce endoplasmic reticulum stress in INS-1-3 cells. Digital gene expression profiling technique was used to detect differentially expressed gene. The changes of gene expression were detected by expression pattern clustering analysis, gene ontology (GO) function and pathway enrichment analysis. Real time polymerase chain reaction (RT-PCR) was used to verify the key changes of gene expression. Results: Compared with the control group, there were 57 (45 up-regulated, 12 down-regulated) and 135 (99 up-regulated, 36 down-regulated) differentially expressed genes in TG and TM group, respectively. GO function enrichment analyses indicated that the main enrichment was in the endoplasmic reticulum. In signaling pathway analysis, the identified pathways were related with endoplasmic reticulum stress, antigen processing and presentation, protein export, and most of all, the maturity onset diabetes of the young (MODY) pathway. Conclusion: Under the condition of endoplasmic reticulum stress, the related expression changes of transcriptional factors in MODY signaling pathway may be related with the impaired function in islet beta cells.

[Clinical features and scalp video electroencephalogram analysis of patients with insular epilepsy].

Objective: To explore the clinical and scalp video electroencephalographic(EEG) characteristics of insula lobe epilepsy identified by intracranial electrode, and to provide some references for early diagnosis and clinical evaluation in patients with insula lobe epilepsy. Methods: A total of 12 patients diagnosed with insula lobe epilepsy identified by intracranial electrode were included from Xuanwu Hospital of Capital Medical University from January 2013 to December 2015.Their clinical and EEG data were analyzed. Results: 91.7% of patients had sensory aura before seizure, and had clear consciousness at onset initial time.Visceral motor and visceral sensation were the most common symptoms, and the epilepsy was always ended with somatic movement.There was no prominent characteristic changes in scalp EEG. Conclusions: Clinical features are important for insula lobe epilepsy diagnosis.When patients present symtoms aforementioned, insula lobe epilepsy should be considered.

[Myoclonus and it's associated factors in Creutzfeldt-Jakob disease].

Objective: To analyze features and the related factors of myoclonus of 47 patients with probable or possible Creutzfeldt-Jakob disease (CJD). Methods: All patients diagnosed with"suspected CJD" and hospitalized in Xuanwu Hospital from January 2013 to November 2015 were included, and their clinical information and myoclonus features were analyzed. Age, clinical, course and manifestation, EEG, MRI, CSF14-3-3 features between myoclonus positive group and negative group were compared, and the correlation between myoclonus features and these factors was analyzed using Spearman correlation analyses. Results: (1) Occurrence rate of extrapyramidal symptoms (P=0.028), visual impairment (P=0.025) and dyssomnia (P=0.004) were higher in myoclonus positive group, the differences were significant. Spearman correlation analysis showed that myoclonus was related to extrapyramidal symptoms (P=0.024), visual impairment (P=0.030) and dyssomnia (P=0.001). (2) EEG features showed no significant difference between myoclonus positive and negative group. The 17 myoclonus positive patients were divided into three subgroups, typical EEG change group 52.94%(9/17), atypical EEG change group 23.53%(4/17) and no EEG change group 23.53%(4/17). Difference of myoclonus and other clinical manifestations were not significant among the three subgroups(P>0.05); correlation analysis also found no statistically significant correlation between myoclonus and EEG (P=0.201). Conclusions: Myoclonus often occurs after the damage of locomotor system (including pyramidal tract, extracorticospinal tract and cerebellum) among CJD patients, and it is related to extrapyramidal symptoms , visual impairment and dyssomnia . There is undefined correlation between myoclonus and periodic sharp wave complexes (PSWC) in EEG.

Inhibition of gap junctions relieves the hepatotoxicity of TNF-α.

The aim of this study was to observe the influence of gap junction (GJ) functional changes on the hepatotoxicity of TNF-α. Three different methods were employed to study functional effects of the GJ inhibition: 1) pretreatment with a GJ inhibitor; 2) inoculation of cells at high and low densities; and 3) inhibition of the expression of connexin 32 (Cx32) by small inhibitory RNA transfection. We then observed the influence of these treatments on hepatotoxicity following treatment with different concentrations of TNF-α for various duration. The hepatotoxicity of TNF-α was observed to occur in a dose- and time-dependent manner; after pretreatment inhibition, the hepatotoxicity of TNF-α was significantly reduced (P < 0.01). The hepatotoxicity of TNF-α was also found to be remarkably lower in cells that had been inoculated at low density (as measured by the amount of GJ formation among cells) than in those inoculated at density (P < 0.01). In addition, following Cx32 inhibition, the hepatotoxicity of TNF-α was significantly decreased (P < 0.01) as well. Together, these results suggest that inhibition of GJ function or of its component Cx32 significantly decreases the hepatotoxicity of TNF-α, and that the expression of Cx32 plays an important role in the hepatotoxicity of TNF-α.

Value of normalized apparent diffusion coefficient for estimating histological grade of vesical urothelial carcinoma.

AIM: To compare the efficacy of apparent diffusion coefficient (ADC) and normalized ADC (nADC) for estimating the histological grade of vesical urothelial carcinoma and to identify an optimal reference for nADC calculation. MATERIALS AND METHODS: Thirty patients with histologically confirmed vesical urothelial carcinomas underwent preoperative diffusion-weighted magnetic resonance imaging (DW-MRI) of the pelvis. nADC of the tumour was calculated as ADC (tumour)/ADC (reference) using urine in the bladder lumen, and the obturator internus and gluteus maximus muscles as reference. Receiver operating characteristic (ROC) curves were constructed and compared to identify an optimal reference for nADC calculation. RESULTS: Both ADC and nADC of low-grade tumours (1.112 ± 0.159 × 10(-3) mm(2)/s, 0.403 ± 0.047 × 10(-3) mm(2)/s) were significantly (p < 0.001) higher than those of high-grade tumours (0.772 ± 0.091 × 10(-3) mm(2)/s, 0.276 ± 0.033 × 10(-3) mm(2)/s). The area under the nADC ROC curve using urine as reference was significantly (p = 0.000) larger (0.995) than those using obturator internus (0.960) and gluteus maximus (0.945). CONCLUSIONS: nADC is superior to ADC for estimating the histological grade of bladder carcinoma using urine in the bladder lumen as an optimal reference for nADC calculation.

Changes in the PD-1 and PD-L1 expressions of splenic dendritic cells in multiple-organ dysfunction syndrome mice and their significance.

The aim of this study was to evaluate the expression of surface molecules in splenic dendritic cells (DC) in multiple-organ dysfunction syndrome (MODS) mice and their effects on the immunosuppression of sepsis and MODS. One hundred thirty C57BL/6 mice were divided into 7 groups: 6, 12, 24, 48 h, 5-7 days, 10-12 days, and the normal control group. The sepsis-MODS mouse model was established by zymson injection into the peritoneal cavity. Histopathological changes in the spleen were evaluated by hematoxylin and eosin (HE) staining. After enrichment with BDTM IMag, the expressions of PD-1, PD-L1, MHC-II (I-A(b)), and CD86 in splenic DCs were examined by flow cytometry, and their relationship with sepsis development and MODS was analyzed. The histological structures of the spleen were damaged in the 24-, 48-h, and 10-12-day groups. PD-L1 expression increased 6 h after zymosan injection, decreased to normal levels at 24 and 48 h, and increased at 5-7 days, peaking at 10-12 days. The change in PD-1 expression roughly paralleled that of PD-L1. MHC-II and CD86 increased at 6 and 12 h, and dropped to normal levels at 10-12 days. In the early stage of injury, splenic DCs were mainly activated, whereas in the later stage, the expressions of the negative co-stimulatory molecules, PD-L1 and PD- 1, were upregulated, similar to tolerogenic DCs. Splenic DCs might suppress the stimulation of T lymphocytes in MODS mice through the PD-L1/PD-1 pathway, which would induce immunosuppression and the pathogenesis of MODS.

Inhibition effect of silver nanoparticles on herpes simplex virus 2.

The herpes simplex virus 2 (HSV-2) is one of the most important sexually transmitted pathogens, and can facilitate the spread of human immunodeficiency virus. The currently available antiviral drugs have certain limitations. Nanosilver has received increasing attention recently with respect to its antibacterial and antiviral properties. The purpose of this study was to determine the inhibiting effect and mechanism of silver nanoparticles (Ag-NPs) on HSV-2. The cytotoxicity of Vero cells induced by different Ag-NP concentrations was investigated by using the methyl thiazolyl tetrazolium (MTT) assay. The inhibiting effect of Ag-NPs on HSV-2 at various times was also evaluated by using a plaque assay. The toxicity of 100 µg/mL Ag-NPs on Vero cells was very low. The mixture of Ag-NP suspension and HSV-2 prior to infecting cells could significantly inhibit the production of progeny viruses. Ag-NPs also inhibited the replication of HSV-2 for 24 h before infecting cells with HSV-2. Therefore, 100 µg/mL Ag-NPs could completely inhibit HSV-2 replication. Ag-NPs at nontoxic concentrations were capable of inhibiting HSV-2 replication when administered prior to viral infection or soon after initial virus exposure. This suggests that the mode of action of Ag-Nps occurs during the early phases of viral replication.

[Related factors of acute symptomatic seizures and epilepsy in children with myelin oligodendrocyte glycoprotein antibody-associated disease].

Objective: To investigate the risk factors of acute symptomatic seizures (ASS) and epilepsy in children with myelin oligodendrocyte glycoprotein antibody-associated disease (MOGAD). Methods: A ambispective cohort study was used including 74 children with MOGAD who were admitted to the Department of Pediatrics of Peking University First Hospital from January 2013 to June 2023 and were followed up. Demographic information, clinical information, treatment status, ASS and epilepsy status were collected. The clinical phenotypes were classified. According to the presence or absence of ASS in the course of disease, the children and the course of disease were divided into groups with and without ASS. Chi-square test, Fisher exact test and Mann Whitney U test were used to analyze the correlation between symptoms and auxiliary examination characteristics and the occurrence of ASS in the two groups of children. Multivariate Logistic regression analysis was used for multivariate analysis. Results: The onset age of the 74 children with MOGAD was 6.58 (3.80, 9.67) years, including 38 females (51.4%) and 36 males (48.6%). The duration of the final follow-up was 2.67 (1.10, 4.12) years, with a total of 239 times acute clinical episodes. ASS occurred in 39.2% (29/74) children during the course of disease and in 29.3% (70/239) of attacks. The common phenotypes were ADEM (67 times (28.0%)), optic neuritis (37 times (15.4%)) and cerebral cortical encephalitis (31 times (13.0%)) in 239 times acute clinical episodes. The incidence of ASS in ADEM and cerebral cortical encephalitis phenotype was 28.4%(19/67) and 100.0% (31/31), respectively. Multivariate analysis showed that cortical involvement on magnetic resonance imaging during clinical attacks was an independent risk factor for ASS (ß=-1.49, OR=0.23) after excluding attacks involving only optic nerve or spinal cord (49 episodes). During the follow-up, 5 children (6.8%) had epilepsy, and all children with epilepsy had multiple clinical attacks of MOGAD and previous ASS. Conclusions: Cortical involvement on magnetic resonance imaging during clinical episodes is an independent risk factor for ASS in children with MOGAD. All MOGAD children with epilepsy had ASS and multiple MOGAD clinical episodes in the past.

Two ground-state bright solitons in fractional-order spin-orbit-coupled Bose-Einstein condensates.

We study two types of bright solitons in zero and non-zero detuning spin-orbit-coupled (SOC) Bose-Einstein condensates in fractional effect by variational and imaginary-time evolution method. The results show that the variation of SOC with fractional kinetic energy operator affects the existence and form of solitons. In particular, we found that SOC strengthγ=1.0could be a transition point. In other words, forγ<1.0andγ⩾1.0, the soliton states and pseudo-spin polarization in non-zero detuning show opposite changes. In addition, moving bright solitons with zero and non-zero detuning are studied.

Variability in chewing, ruminal fermentation, digestibility and bacterial communities between subacute ruminal acidosis-susceptible and acidosis-tolerant sheep.

Sorting behaviour is a common phenomenon observed in ruminants when they are provided with a total mixed ration, which contributes to variations in the severity of subacute ruminal acidosis (SARA). Pelleted total mixed ration (PTMR) reduces sorting, but high-grain content increases acidosis risk. However, whether the variability in the severity of SARA exists in sheep fed the same high-grain PTMR is less understood. This study aimed to investigate SARA variability among individual sheep offered a high-grain PTMR, considering chewing activity, ruminal fermentation, bacterial communities and nutrient digestibility. Twenty ruminally cannulated male Hu sheep were individually housed in cages and fed a PTMR comprising 80% concentrate mix and 20% roughage. A 14-day adaptation period to the diet and facilities was provided before a 10-day sample collection period. Continuous monitoring of ruminal pH was conducted for 48 h, during which time chewing activity was also recorded. Ruminal fluid samples were collected for analysis of volatile fatty acid and microbial DNA extraction. Faecal samples were collected to measure nutrient digestibility. Based on their acidosis index, the sheep were classified into two groups: SARA-susceptible group (n = 6) and SARA-tolerant group (n = 6). The SARA-susceptible sheep exhibited a lower ruminal mean pH and minimum pH than the SARA-tolerant sheep (P < 0.05). Additionally, the SARA-susceptible group increased the acidosis index, duration and areas of pH below 5.8 and 5.6 compared to the SARA-tolerant group (P < 0.05). The SARA-susceptible group also exhibited a longer ruminating time than the SARA-tolerant group (P < 0.05). The SARA-susceptible group exhibited a tendency to increase the relative abundance of Firmicutes (P = 0.089), while simultaneously decreasing the copy number of Fibrobacter succinogenes in the rumen, as well as the digestibility of NDF and ADF compared to the SARA-tolerant group (P < 0.05). The acidosis index was found to be positively correlated with ruminating time (min/kg DM intake (DMI)) and total chewing time (min/kg DMI), but negatively correlated with the copy number of Fibrobacter succinogenes and Ruminococcus albus in the rumen. These findings indicate that there exists variability in the SARA severity among sheep when fed a high-grain PTMR, as evidenced by varied chewing activity, bacterial communities and nutrient digestibility. Ruminating time, total chewing time per kilogram of DMI as well as the copy number of Fibrobacter succinogenes and Ruminococcus albus in the rumen hold potential as indicators for assessing the severity of SARA.

Preliminary Extracellular Vesicle Profiling in Drainage Fluid After Neck Dissection in OSCC.

Lymph node metastasis is related to poor prognosis in oral squamous cell carcinoma (OSCC), and few studies have explored the relevance of postoperative drainage fluid (PDF) in metastasis. Extracellular vesicles (EVs) are nanosized vesicles that can transfer oncogenic molecules to regulate tumorigenesis. However, the proteomic profile of postoperative drainage fluid-derived EVs (PDF-EVs) in OSCC has not been elucidated. Herein, we collected drainage fluid from OSCC patients after neck dissection to investigate the difference in PDF-EVs between patients with metastatic lymph nodes (the LN+ group) and nonmetastatic lymph nodes (the LN- group). The proteomic profile of PDF-EVs from the LN+ and LN- groups was compared using label-free liquid chromatography tandem-mass spectrometry-based protein quantification. The results revealed that PDF-EVs were mainly derived from epithelial cells and immune cells. A total of 2,134 proteins in the PDF-EVs were identified, and 313 were differentially expressed between the LN+ and LN- groups. Metabolic proteins, such as EHD2 and CAVIN1, were expressed at higher levels in the LN+ group than in the LN- group, and the levels of EHD2 and CAVIN1 in the postoperative drainage fluid were positively correlated with lymph node metastasis. Our study revealed previously undocumented postoperative drainage fluid-associated proteins in patients with metastatic OSCC, providing a starting point for understanding their role in metastatic and nonmetastatic OSCC.

A minimized and efficient low temperature loading device for indentation.

A minimized and efficient low temperature loading device cooling with Peltier coolers for indentation test is developed. Both specimen and indentation tip are immersed in 50% methanol-water solution, totally eliminating the contact thermal drift problem. Low temperature indentation tests down to 253.8 K can be realized within 10 min. The size of the device is miniaturized within 50 × 40 × 30 mm3, and no vacuum environment is required. Monocrystalline copper is tested to perform the feasibility of the device. Based on the simple structure and stable experimental effect, the developed device can be integrated into various types of current indentation devices to attach low temperature testing ability.

[Effects and mechanism of human umbilical vein endothelial cells-derived exosomes on wound healing in diabetic rabbits].

Objective: The investigate the effects and mechanism of exosomes derived from human umbilical vein endothelial cells (HUVECs) on wound healing in diabetes rabbits. Methods: The experimental research methods were used. The primary vascular endothelial cells (VECs) and human skin fibroblasts (HSFs) were extracted from skin tissue around ulcer by surgical excision of two patients with diabetic ulcer (the male aged 49 years and the female aged 58 years) admitted to Xiangya Third Hospital of Central South University in June 2019. The cells were successfully identified through morphological observation and flow cytometry. The HUVEC exosomes were extracted by ultracentrifugation and identified successfully by morphological observation, particle size detection, and Western blotting detection. Twenty female 3-month-old New Zealand rabbits were taken to create one type 2 diabetic full-thickness skin defect wound respectively on both sides of the back. The wounds were divided into exosomes group and phosphate buffer solution (PBS) group and treated accordingly, with 20 wounds in each group, the time of complete tissue coverage of wound was recorded. On PID 14, hematoxylin-eosin staining or Masson staining was performed to observe angiogenesis or collagen fiber hyperplasia (n=20). The VECs and HSFs were co-cultured with HUVEC exosomes for 24 h to observe the uptake of HUVEC exosomes by the two kinds of cells. The VECs and HSFs were divided to exosome group treated with HUVEC exosomes and PBS group treated with PBS to detect the cell proliferation on 4 d of culture with cell count kit 8, to detect and calculate the cell migration rate at 24 and 48 h after scratch by scratch test, to detect the cell migration number at 24 h of culture with Transwell test, and to detect the mRNA expressions of nuclear factor-erythroid 2-related factor 2 (NRF2) and transcription activating factor 3 (ATF3) by real time fluorescence quantitative reverse transcription polymerase chain reaction. Besides, the number of vascular branches and vascular length were observed in the tube forming experiment after 12 h of culture of VECs (n=3). The VECs and HSFs were taken and divided into PBS group and exosome group treated as before, and NRF2 interference group, ATF3 interference group, and no-load interference group with corresponding gene interference. The proliferation and migration of the two kinds of cells, and angiogenesis of VECs were detected as before (n=3). Data were statistically analyzed with analysis of variance for repeated measurement, one-way analysis of variance, independent sample t test, and least significant difference test. Results: The time of complete tissue coverage of wound in exosome group was (17.9±1.9) d, which was significantly shorter than (25.2±2.3) d in PBS group (t=4.54, P<0.05). On PID14, the vascular density of wound in PBS group was significantly lower than that in exosome group (t=10.12, P<0.01), and the collagen fiber hyperplasia was less than that in exosome group. After 24 h of culture, HUVEC exosomes were successfully absorbed by VECs and HSFs. The proliferative activity of HSFs and VECs in exosome group was significantly higher than that in PBS group after 4 d of culture (with t values of 54.73 and 7.05, respectively, P<0.01). At 24 and 48 h after scratch, the migration rates of HSFs (with t values of 3.42 and 11.87, respectively, P<0.05 or P<0.01) and VECs (with t values of 21.42 and 5.49, respectively, P<0.05 or P<0.01) in exosome group were significantly higher than those in PBS group. After 24 h of culture, the migration numbers of VECs and HSFs in exosome group were significantly higher than those in PBS group (with t values of 12.31 and 16.78, respectively, P<0.01). After 12 h of culture, the mRNA expressions of NRF2 in HSFs and VECs in exosome group were significantly higher than those in PBS group (with t values of 7.52 and 5.78, respectively, P<0.05 or P<0.01), and the mRNA expressions of ATF3 were significantly lower than those in PBS group (with t values of 13.44 and 8.99, respectively, P<0.01). After 12 h of culture, the number of vascular branches of VECs in exosome group was significantly more than that in PBS group (t=17.60, P<0.01), and the vascular length was significantly longer than that in PBS group (t=77.30, P<0.01). After 4 d of culture, the proliferation activity of HSFs and VECs in NRF2 interference group was significantly lower than that in PBS group and exosome group (P<0.05 or P<0.01); the proliferation activity of HSFs and VECs in ATF3 interference group was significantly higher than that in PBS group (P<0.05 or P<0.01) and significantly lower than that in exosome group (P<0.05 or P<0.01). At 24 and 48 h after scratch, the migration rates of HSFs and VECs in ATF3 interference group were significantly higher than those in PBS group (P<0.05 or P<0.01) and significantly lower than those in exosome group (P<0.05 or P<0.01). At 24 and 48 h after scratch, the migration rates of HSFs and VECs in NRF2 interference group were significantly lower than those in PBS group and exosome group (P<0.05 or P<0.01). After 24 h of culture, the migration numbers of VECs and HSFs in ATF3 interference group were significantly more than those in PBS group (P<0.05) and significantly less than those in exosome group (P<0.05 or P<0.01); the migration numbers of VECs and HSFs in NRF2 interference group were significantly less than those in PBS group and exosome group (P<0.01). After 12 h of culture, the vascular length and number of branches of VECs in NRF2 interference group were significantly decreased compared with those in PBS group and exosome group (P<0.01); the vascular length and number of branches of VECs in ATF3 interference group were significantly increased compared with those in PBS group (P<0.01) and were significantly decreased compared with those in exosome group (P<0.01). Conclusions: HUVEC exosomes can promote the wound healing of diabetic rabbits by promoting the proliferation and migration of VECs and HSFs, and NRF2 and ATF3 are obviously affected by exosomes in this process, which are the possible targets of exosome action.

[Clinical follow-up study of myelin oligodendrocyte glycoprotein antibody-associated disease in children].

Objective: To summarize the clinical characteristics of myelin oligodendrocyte glycoprotein antibody-associated disease (MOGAD) and compare the differences in efficacy of different disease-modifying drugs. Methods: An ambispective cohort study was conducted in 42 children diagnosed with MOGAD at Department of Pediatrics, Peking University First Hospital from January 2012 to March 2021 and conducted long-term follow-up to analyze clinical phenotypes and compare the efficacy of different disease-modifying drugs such as rituximab, mycophenolate mofetil and azathioprine. Kruskal-Wallis H test was used to compare the annual relapse rate of disease-modifying drugs at different times, expanded disability status scale (EDSS) score at the last follow-up, and Wilcoxon rank test was used to compare the annual relapse rate before and after modified disease therapy. The Log-rank (Mantel-Cox) survival curve was used to compare the relapse rate of different disease-modifying drugs. Results: Of the 42 cases, 22 were male and 20 were female, with the age at disease onset of 5.96 (2.33-12.90) years. The disease duration was 4.46 (1.25-13.00) years at the last follow-up with 161 clinical acute attacks. Acute disseminated encephalomyelitis (ADEM) was the most common phenotype of first attack and all attacks during disease course ((60% (25/42) for first attack, 38% (61/161) for all attacks). The most common clinical syndrome was neuromyelitis optica spectrum disorders (NMOSD) (50%, 21/42). Of the 42 children, 5 (12%) showed encephalitis and 6 (14%) combined with anti-N-methyl-D-aspartate receptor (NMDAR) antibody overlap syndrome. The most commonly involved areas of brain magnetic resonance imaging (MRI) were subcortical white matter (71%, 88/124), cortex (26%, 32/124) and periventricular white matter (25%, 32/124). Spinal cord MRI was most frequently involved in cervical (70%, 16/23) and thoracic (61%, 14/23) medulla, and 43% (10/23) longitudinally extensive transeverse myelitis. Disease-modifying drugs were used in 34 patients. The annual relapse rate after treatment with rituximab, mycophenolate mofetil and azathioprine decreased (all P<0.05) and there was no statistically significant difference in the annual relapse proportion among the groups (P=0.307). Conclusions: The most common clinical attack of first and all of MOGAD in children is ADEM, and the most common clinical syndrome is NMOSD. Rituximab, mycophenolate mofetil and azathioprine can reduce the annual relapse rate, but it is not clear effect of which treatment is better.

Characterization of a cis-acting regulatory element which silences expression of the class II-A beta gene in epithelium.

Class II major histocompatibility complex (MHC) genes encode for alpha/beta chain pairs that are constitutively expressed principally on mature B cells and dendritic cells in mice. These gene products are easily induced on macrophages with cytokines, and may also aberrantly appear on the surface of epithelium during immune injury. The appearance of class II determinants in parenchymal tissue potentially renders these somatic cells capable of antigen presentation to circulating CD4+ T lymphocytes, and their absence may be protective for normal tissues expressing self-antigens. The low surface class II expression observed on parenchymal cells generally correlates with low levels of mRNA, suggesting that transcription rate is a major element in class II regulation. To understand the transcriptional mechanism maintaining low basal surface expression of class II in somatic cells, we transiently transfected mini-gene reporter constructs to study the regulation of the murine A beta promoter in a cultured renal epithelial cell line. We describe here a negative cis-acting regulatory region located between -552 and -489 bp upstream of the A beta cap site that silences the transcriptional activity of the A beta promoter in epithelial cells in an orientation-dependent manner, and is also able to silence a heterologous promoter. This region is not active in class II-expressing B cells (BAL-17) in culture, but is functional in two other murine class II-negative cell lines, fibroblasts and thymoma T cells. Using competition electrophoretic mobility shift assays, we have localized the core protein binding site within this region to an 8-10-bp response element, designated A beta NRE, at -543 to -534 bp. A nuclear extract from BAL-17 cells does not bind to this element. Mutation of this site abrogates the transcriptional silencing activity of the region. We conclude that the transcription of class II-A beta in parenchymal cells, and some lymphocytes, can be actively repressed by an upstream silencing element.