Plasminogen degrades α-synuclein, Tau and TDP-43 and decreases dopaminergic neurodegeneration in mouse models of Parkinson's disease.

Parkinson's disease (PD) is the second most frequently diagnosed neurodegenerative disease, and it is characterized by the intracellular and extracellular accumulation of α-synuclein (α-syn) and Tau, which are major components of cytosolic protein inclusions called Lewy bodies, in the brain. Currently, there is a lack of effective methods that preventing PD progression. It has been suggested that the plasminogen activation system, which is a major extracellular proteolysis system, is involved in PD pathogenesis. We investigated the functional roles of plasminogen in vitro in an okadaic acid-induced Tau hyperphosphorylation NSC34 cell model, ex vivo using brains from normal controls and methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-treated mice, and in vivo in a widely used MPTP-induced PD mouse model and an α-syn overexpression mouse model. The in vitro, ex vivo and in vivo results showed that the administered plasminogen crossed the blood‒brain barrier (BBB), entered cells, and migrated to the nucleus, increased plasmin activity intracellularly, bound to α-syn through lysine binding sites, significantly promoted α-syn, Tau and TDP-43 clearance intracellularly and even intranuclearly in the brain, decreased dopaminergic neurodegeneration and increased the tyrosine hydroxylase levels in the substantia nigra and striatum, and improved motor function in PD mouse models. These findings indicate that plasminogen plays a wide range of pivotal protective roles in PD and therefore may be a promising drug candidate for PD treatment.

Toxic effects of atmospheric deposition in mining areas on wheat seedlings.

Storage and transportation of coal, as well as operation of coal-fired power plants, produce amounts of metallic exhaust that may lead to different atmospheric environment in the overlapped areas of farmland and coal resource (OAFCR) environment. To investigate the effects of different atmospheric environment in the OAFCR region (north of Xuzhou) on wheat seedlings (AK-58), a box experiment was conducted and compared to an area far from the OAFCR (south of Xuzhou). The study revealed that (1) compared to the southern suburb of Xuzhou, the fresh and dry weight, activities of photosynthetic enzymes and POD of wheat seedlings in the OAFCR reduced obviously. (2) Significantly higher levels of Cr, Cd, Pb, Zn, and Cu were found in the shoots and roots of wheat seedlings in the OAFCR, with lower transfer factor for heavy metals (except Cd and As) in comparison to those in the southern suburb. And the bioconcentration factors of heavy metals (except As) in wheat seedlings in the OAFCR were significantly higher. (3) Nearly 90% of heavy metals (Pb, Cu, Cd, Zn, and Cr) absorbed by wheat were stored in cell walls and soluble fractions, with significantly higher contents of Cu and Cr in wheat seedlings' cell walls and higher contents of Pb, Zn, and Cd in soluble components found in the OAFCR. Our results showed that atmospheric deposition in the mining area has a certain toxic effect on wheat seedlings, and this study provides a theoretical basis for OAFCR crop toxicity management.

Plasminogen decreases Aß42 and Tau deposition, and shows multi-beneficial effects on Alzheimer's disease in mice and humans.

Alzheimer's disease (AD) is the most common neurodegenerative disorder in the world. The aggregation of both amyloid beta (Aß) peptides extracellularly and Tau proteins intracellularly plays key roles in the pathological consequences of AD, which lead to cholinergic neurodegeneration and eventually death. Currently, there are no effective methods to stop the progression of AD. Using ex vivo, in vivo and clinical approaches, we investigated the functional effects of plasminogen on the widely used FAD, Aß42 oligomer or Tau intracranial injection-induced AD mouse model and explored its therapeutic effects on patients with AD. The results show that intravenously injected plasminogen rapidly crosses the blood‒brain barrier (BBB); increases plasmin activity in the brain; colocalizes with and effectively promotes the clearance of Aß42 peptide and Tau protein deposits ex vivo and in vivo; increases the choline acetyltransferase (ChAT) level and decreases the acetylcholinesterase (AChE) activity; and improves the memory functions. Clinically, when GMP-level plasminogen was administered to 6 AD patients for 1-2 weeks, their average scores on the Minimum Mental State Examination (MMSE), which is a standard scoring system used to measure the memory loss and cognitive deficits, were extremely significantly improved by 4.2 ± 2.23 points, e.g., an average increase from 15.5 ± 8.22 before treatment to 19.7 ± 7.09 after treatment. The preclinical study and pilot clinical study suggest that plasminogen is effective in treating AD and may be a promising drug candidate.

Questionnaire survey of risk factors for recurrence of ocular inflammation in patients with uveitis after SARS-CoV-2 infection.

Introduction: During the COVID-19 pandemic in China, the proportion of patients with uveitis who were infected with SARS-CoV-2 increased greatly. The impact of SARS-CoV-2 infection on patients with uveitis has not been fully described. Methods: A questionnaire on SARS-CoV-2 infection was sent to patients with uveitis to assess ocular and systemic conditions before and after infection. Chi-square analysis and multifactorial regression analysis were used to investigate the associations between each risk factor and the recurrence of uveitis after SARS-CoV-2 infection. Results: One hundred thirty-nine patients with noninfectious uveitis completed the questionnaire; 114 (82.0%) had COVID-19, and 27 (23.7%) had recurrent or exacerbated uveitis after COVID-19. There was a higher rate of recurrence or aggravation of ocular inflammation in patients who developed severe COVID-19 symptoms (severe group 8/20 vs. nonsevere group 19/94). There were significant differences in the rates of recurrence and aggravation between the two groups of patients who differed in terms of ocular inflammatory activity within 3 months prior to SARS-CoV-2 infection (χ2 = 10.701, P=0.001), as well as in the rates of recurrence and aggravation after cessation of systemic immunomodulatory therapy. After multifactorial regression analysis, patients with active ocular inflammation within 3 months prior to SARS-CoV-2 infection had a greater risk of recurrence or exacerbation of uveitis after COVID-19 (OR=4.298, P=0.002). Conclusion: The degree of ocular inflammatory activity within 3 months prior to SARS-CoV-2 infection may be a major factor influencing the recurrence or exacerbation of uveitis after infection. Interruption of medication should be minimized in patients with unstable inflammatory control.

Epidemiological and genetic characteristics of human metapneumovirus in pediatric patients across six consecutive seasons in Beijing, China.

OBJECTIVES: To investigate the genetic characteristics of human metapneumovirus (hMPV) circulating among children with acute respiratory tract infections (ARTIs) in Beijing, China. METHODS: Clinical samples were obtained from outpatients and hospitalized children with ARTIs between August 2010 and July 2016. Reverse transcription polymerase chain reaction assays were used to screen and identify hMPV, while partial glycoprotein gene sequences were used for phylogenetic analysis. RESULTS: Among the 10 918 samples, 292 (2.7%) were positive for hMPV. Overall, the virus was more prevalent among inpatients (4.3%) than outpatients (1.2%). A biennial alternating pattern of hMPV infection was observed, with infection rates fluctuating between 1.6% and 4.0%. Most cases were detected between December and April, showing clear-cut seasonality. Sub-genotypes A2b, B1, and B2 co-circulated in winter and spring in an alternating pattern, while only one A1-positive case was observed in 2012. The seasonal peak of hMPV was slightly delayed or overlapped with that of respiratory syncytial virus and influenza virus. hMPV activity increased in the 2010-2011 and 2014-2015 seasons, when influenza activity was apparently decreased compared with other epidemic seasons. CONCLUSIONS: This study provides information on the epidemiological and genetic characteristics of hMPV in children in Beijing, and reinforces the significance of hMPV in children with ARTIs, especially lower respiratory tract infections.

Clinical Profile, Ultra-Wide-Field Fluorescence Angiography Findings, and Long-Term Prognosis of Uveitis in Tubulointerstitial Nephritis and Uveitis Syndrome at One Tertiary Medical Institute in China.

Purpose: To analyze the clinical features and long-term prognosis of uveitis in tubulointerstitial nephritis and uveitis (TINU) syndrome in a cohort of Chinese patients. Methods: A total of 32 patients with TINU syndrome between 2000 - 2016 were analyzed retrospectively. Results: Mild anterior inflammation was observed in all cases. Ultra-wide-field fluorescence angiography (UWFA) was conducted on 13 patients, and peripheral vascular leakage was observed in 22/26 eyes (84.62%) compared with active anterior chamber inflammation in 13/26 eyes (50%). Three patients received increased corticosteroid dosage therapy on their first episode of uveitis guided by UWFA had stopped steroids without recurrence. Conclusion: The uveitis in TINU syndrome was mild. Corticosteroids were efficient in most cases, but a slower tapering and long-term treatment were required. UWFA is sensitive in detecting the activity of uveitis and might be useful in monitoring disease progression. Complete withdrawal of medication was difficult to achieve in some cases.

A Cross-Sectional Observational Study of Nailfold Capillary Morphology in Uveitis.

PURPOSE: We performed nailfold capillary microscopy to explore microvasculature abnormalities in uveitis overall and uveitis stratified in various ways. METHODS: This was a cross-sectional, case-control, observational study. One hundred and seven uveitis patients and 130 control subjects were included. We used a JH-1004 capillaroscope to perform nailfold capillary video microscopy on the fourth and fifth digits of each subject's nondominant hand. Videos were evaluated for hemorrhages, dilated capillary loops > 25 µm, and avascular zones > 200 µm. Univariate analyses were used for the assessment of case-control morphological differences and multivariate analyses were performed to assess the relation between nailfold capillaroscopic findings and uveitis subgroups. RESULTS: In univariate analysis, uveitis patients were more likely to have higher tortuosity ratings and reduced capillary density compared to controls (p < 0.001 for both); furthermore, dilated capillary loops, avascular zone and hemorrhages were more frequent in uveitis versus control subjects (p < 0.001 for all). Among cases, every unit increase in capillary density (vessels/mm) was associated with active uveitis (n = 72 cases) versus inactive disease (n = 35 cases; odds ratio (OR) = 1.7; (95% confidence interval (CI), 1.2-2.5) in multivariate analysis. Furthermore, the presence of any nailfold hemorrhage versus the absence of hemorrhage was more likely to be associated with posterior and panuveitis (n = 41 cases combined) compared to anterior and intermediate uveitis (n = 66 cases combined; OR = 5.8; 95% CI, 2.3-14.2). Moreover, we found a positive correlation between peripheral retinal leakage and nailfold capillaries dilation (r = 0.33; p = 0.015) that was not strictly significant based on the number of comparisons made. CONCLUSIONS: Our study provides support for non-ocular capillary bed abnormalities in uveitis, with interesting correlations based on disease stage and anatomical classification.

Subretinal echinococcosis: a case report.

BACKGROUND: Echinococcosis is a dangerous zoonotic parasitic disease. Ocular echinococcosis is very rare, especially the hydatid cysts in subretinal space. We present a case of subretinal echinococcosis and management. CASE PRESENTATION: A 37-year-old man with subretinal echinococcosis who developed panuveitis and visual impairment. The patient lives on agriculture and animal husbandry, which made him susceptible to parasitic infection. He had severe panuveitis and blurred vision on arrival at hospital. According to his ocular examination and systemic review, the subretinal echinococcosis diagnosis was made. The patient received pars plana lensectomy and pars plana vitrectomy. The lesion underneath his retina was removed, and histopathology examination confirmed the subretinal echinococcosis diagnosis. CONCLUSIONS: Echinococcosis is a dangerous zoonotic parasitic disease in pastoral areas. Ocular echinococcosis is usually secondary to systemic infection. Although the incidence is rare, the disease could lead to destructive visual function impairment.

[Whole Genome Sequencing and Phylogenetic Analyses of Sub-genotype A1 of the Human Metapneumovirus Detected in an Infant with Pneumonia].

The human metapneumovirus (HMPV) is an important pathogen in respiratory-tract infections in children. We undertook genomic sequence analyses and described the genetic characteristics of an uncommon sub-genotype, the HMPV A1 strain, and provide useful data for further studies. The HMPV A1(BJ-1610)strain was identified from a nasopharyngeal aspirate collected from a 3-month-old female with bronchopneumonia. Gene fragments of BJ-1610 were amplified by reverse transcription-polymerase chain reaction(RT-PCR)and assembled by DNAStar software. Sequence alignment for BJ-1610 and other HMPV reference strains with four known genotypes available in the GenBank database was conducted by DNAStar. Phylogenetic trees were created using MEGA 6.06 software. The whole genome of BJ-1610 was 13406nt in length (GenBank accession number:KU821121).Compared with HMPV reference strains,BJ-1610 shared the highest similarities with HMPV/AUS/150229278/2003/A(KC562226)from Australia, which was classified into sub-genotype A1.The nucleotide identity of the full genome between BJ-1610 and KC562226was 98.4%.N,P,F,M2-2and L genes had great similarity with KC562226 compared with other reference strains, whereas SH and G genes shared higher similarities with other strains of sub-genotype A1.Phylogenetic analyses of the whole genome showed that BJ-1610 was clustered into sub-genotype A1 and was close to KC562226.The N,P,M,F,M2-1,M2-2and L genes of BJ-1610 showed the same genetic features as the whole genome, whereas the variable genes SH and G were closest to KC403980.The F protein of BJ-1610 showed high genetic conservation. The length of the SH protein of BJ-1610 changed from 552 bp to 567 bp due to mutations in the stop codon. The amino-acid mutations on protein G led to a decrease in the number of N-glycosylation sites. As an infrequently circulating genotype, sequence analyses of the whole genome of a HMPV A1strain(BJ-1610)will promote further studies on its epidemiology and pathogenicity, and aid the development of vaccines and antiviral drugs.

[The clinical investigation of peripheral retinal changes of anterior uveitis].

OBJECTIVE: To observe the peripheral retinal changes of anterior uveitis patients using ultra-wide field fluorescein angiography. METHODS: Retrospective case series study. Thirty-three eyes of 20 patients diagnosed as anterior uveitis from September to November 2013 in the ophthalmic clinic of Peking University First Hospital was included. All the patients underwent routine ocular examination, followed by ultra-wide field Fluorescein. RESULTS: There were 13 male patients and 7 female patients, aged 23-66, an average of 46 years old. The anterior uveitis recurrent in both eyes in 13 patients and only one eye in 7 patients. Uveitis duration ranging from months to 30 years. Twenty eyes were at the active stage of the anterior uveitis while 13 eyes in the quiescent stage. Peripheral retinal vessels leakage was detected in 57.6% (19/33) of the eyes, among which in eyes with active disease the rate of peripheral retinal vessels leakage was 75% (15/20), and in eyes with quiescent disease the rate was 30.8% (4/13). Optic disk hyperfluorescence was found in 6 of 20 eyes with active disease. Cystoid macular edema was found in 4 eyes. In addition, peripheral retinal hyperfluorescence dot and zone were detected in 2 eyes. CONCLUSIONS: Peripheral retinal vessels were detected to have inflammative leakage in patients of anterior uveitis by ultra-wide field fluorescein agiography. Peripheral retinal inflammation may still exist even in the quiescent stage of the anterior uveitis.

A prospective, observational study on the application of ultra-wide-field angiography in the evaluation and management of patients with anterior uveitis.

PURPOSE: Evaluation of peripheral retinal vascular changes in anterior uveitis using ultra-wide-field fluorescein angiography. DESIGN: A prospective, observational study of a case series of patients diagnosed with anterior uveitis. SETTING: Clinical observation at an academic medical center. Patient or Study Population: A total of 65 eyes of 33 patients corresponded with the research criteria of anterior uveitis in the opinion of specialists of Peking University First Hospital. OBSERVATION PROCEDURES: Patients were diagnosed primarily through clinical examinations and conventional fluorescein angiography. Subsequently, ultra-wide-field fluorescein angiograms were obtained for each patient. MAIN OUTCOME MEASURES: The main outcome was the detection of peripheral retinal changes by ultra-wide-field fluorescein angiography, and how these changes influenced the evaluation and management of the disease. RESULTS: Peripheral vessel leakage was detected in 27 eyes (42%) with anterior uveitis, of which 15 eyes displayed active inflammation and 12 eyes displayed inactive inflammation. Peripheral vessel leakage was found in seven of eight eyes with cystoid macular edema. Cystoid macular edema was detected in 7 of 27 eyes (26%) with peripheral vessel leakage, whereas 1 of 38 eyes (3%) did not display peripheral vessel leakage (p<0.01). 44.4% of the patients with peripheral vessel leakage had a specific etiology. The relevant treatment strategies were modified based on the results of the ultra-wide-field fluorescein angiography. 12 patients with peripheral vessel leakage and a quiescent anterior segment were added to those receiving topical glucocorticoids, while 3 patients with serious peripheral vessel leakage and an active anterior segment received a sub-Tenon injection of triamcinolone acetonide. CONCLUSIONS: Ultra-wide-field fluorescein angiography was very effective in detecting peripheral retinal vascular pathology in anterior uveitis. The changes found in the periphery were important in the evaluation and management of anterior uveitis.

Synthesis and biological evaluation of quinazoline and quinoline bearing 2,2,6,6-tetramethylpiperidine-N-oxyl as potential epidermal growth factor receptor(EGFR) tyrosine kinase inhibitors and EPR bio-probe agents.

4-anilinoquinazoline and 4-anilinoquinoline scaffolds bearing a 2,2,6,6-tetramethylpiperidine-N-oxyl(TEMPO) have been synthesized and evaluated for their ability to inhibit EGFR tyrosine kinase and A431 cell lines. Compared to their corresponding parent compounds, all of the new compounds bearing a TEMPO showed more efficient inhibition for EGFR and A431 cells. Furthermore, we have proved that these molecules bearing a TEMPO can exactly get into A431 cells exerting inhibitory effect that may be used for EPR detecting. In our docking model, quinazolines bearing a TEMPO on either 6- or 3-positions took different linking modes according to EGFR crystal structure. In contrast to their parent compounds, these new TEMPO-derived analogues possessed compatible inhibitory effect that might be useful as potential EGFR inhibitors and as EPR bio-probes.

Synthesis and biological evaluation of 4-[3-chloro-4-(3-fluorobenzyloxy)anilino]-6-(3-substituted-phenoxy)pyrimidines as dual EGFR/ErbB-2 kinase inhibitors.

A series of 4-[3-chloro-4-(3-fluorobenzyloxy)anilino]-6-(3-substituted-phenoxy)pyrimidine derivatives were elaborately designed based on the skeleton of Lapatinib, and evaluated for their potential to inhibit epidermal growth factor receptor (EGFR) and ErbB-2 tyrosine kinase activities and antiproliferative activities against A431 and SKOV-3 cell lines. Among these synthesized pyrimidine derivatives, 4-[3-chloro-4-(3-fluorobenzyloxy)anilino]-6-(3-acrylamidophenoxy)pyrimidine (6), 4-[3-chloro-4-(3-fluorobenzyloxy)anilino]-6-(3-cyanoacetamidophenoxy)pyrimidine (9), 4-[3-chloro-4-(3-fluorobenzyloxy)anilino]-6-{3-[6-(4-amino)pyrimidinyl]amino) phenoxy}pyrimidine (11) and 4-[3-chloro-4-(3-fluorobenzyloxy)anilino]-6-(3-phenoxyacetamidophenoxy)pyrimidine (14) could significantly inhibit dual EGFR/ErbB-2 kinase activities (IC(50)=37/29 nM, 48/38 nM, 61/42 nM, 65/79 nM, respectively). And compounds 6 and 11 also showed the most potent antiproliferative activities in vitro, with the IC(50) value of 6 being 3.25 µM for A431 and 0.89 µM for SKOV-3, as for 11, 4.24 µM for A431 and 0.71 µM for SKOV-3, respectively. Docking study was also performed to determine the possible binding model.