OBJECTIVE:To investigate antioxidant effects of different extract parts from Aquilaria sinensis leaves. METH-ODS:A. sinensis leaves were extracted by reflux extraction with petroleum ether,ethyl acetate,absolutely ethanol,water to obtain corresponding parts (AGP,AGE,AGA and AGW parts). The ability of different extracts eliminating ABTS+ free radical,DPPH free radical and O2-free radical and reducing Cu2+ were investigated. IC50 was also calculated. Vitamin C(VC)was used as positive control in above tests. RESULTS:Those parts eliminated above free radicals and reduced Cu2+ in concentration-dependant manner, among which maximal elimination rate of AGA and AGW parts to ABTS+ free radical were 100% and 81.07%(IC50=14.89,23.12 mg/L)and higher than that of VC(88%);those of other parts were less than 50%. The maximal elimination rate of AGA parts to DPPH free radical was 67.74%(IC50=47.96 mg/L),but still lower than that of VC(91.44%);the elimination rates of other parts were all weak and lower than 40%. The maximal elimination rates of AGA and AGW parts to O2-free radical were 67.79% and 61.79%(IC50=393.01,421.04 mg/L),but still lower than that of VC(88%);the elimination rates of other parts were all lower than 33%. The maximal reduction percentage of AGP,AGE,AGA and AGW parts to Cu2+ were all higher than 50%(IC50=55.99, 40.27,25.44,31.06 mg/L,respectively),but still lower than that of VC (100%). CONCLUSIONS:Different extract parts from A. sinensis leaves have antioxidant activity in vitro,and that of AGA is the strongest.

BACKGROUNDConstructing replication defective recombinant adenovirus vector expressing the group specific antigen VP6 of human rotavirus and studying the immune responses induced in vivo.

METHODSThe cDNA of full length VP6 was inserted into the adenovirus vector pShuttle-CMV, and recombinant adenovirus genome DNA was obtained through homological recombination in E.coli,then the recombinant adenovirus was gained after transfecting 293 cell line with the genome DNA. Gene integration of VP6 in resultant adenovirus was confirmed by PCR and Southern blot, respectively gene expression was confirmed in 293 cells by Western blot. BALB/c mice were immunized intranasally(inl)and orally(ora), respectively, to test the immunization effects of the adenovirus.

RESULTSRecombinant adenovirus named rvAd-VP6 was obtained. The cDNA of VP6 was integrated in the adenovirus and was able to be expressed in 293 cells stably. The systemic immune responses to rotavirus VP6 could be induced effectively in both oral and intranasal group, the titer of serum IgG antibody in the two group of mice were 1?1 000 and 1?10 000-1?100 000, respectively. In addition to IgG, the serum IgA specific to VP6 could also be detected at a titer of 1?10-1?100. Secretory IgA(sIgA) was detected in both lung lavage fluid and intestinal homogenate when administered intranasally to BALB/c mice, whereas only found in intestinal homogenate in the oral group. The results indicated that the immunization efficacy of intranasal inoculation was superior to that of oral inoculation.

CONCLUSIONSThe recombinant adenovirus vector expressing human rotavirus VP6 was successfully constructed, its ability to induce immune responses has laid a solid foundation for the development of rotavirus genetically engineering vaccine against rotavirus infection.

Adenoviridae ; genetics ; Animals ; Antibodies, Viral ; blood ; Antigens, Viral ; Capsid Proteins ; biosynthesis ; immunology ; Genetic Vectors ; Mice ; Mice, Inbred BALB C ; Recombination, Genetic ; Rotavirus ; immunology

OBJECTIVETo set up a novel, simple, sensitive, specific, repeatable and rapid assay for diagnosis of influenza.

METHODSMonolayers of MDCK cells were inoculated with the specimens for amplifying viral yield, the feature of receptors on cell surface was changed by treatment of neuraminidases of influenza A and B viruses. Afterward, based on the lectin binds to receptors on cell surface with strict specificity,the phenomenon of red blood cell aggregation was observed under the conventional microscope. Finally, the tested results could be determined by the extent of red blood cell aggregation.

RESULTSThere was a complete (%) consistency rate (100%) for viral isolation between new and routine tests. In general, the results were detected with new assay within 20 h. The sensitivity of new assay was over 100-10,000 times higher than that of routine method. Meanwhile, the novel test could not only be used for rapid diagnosis in the clinic, but also be used for influenza surveillance. The best concentration of red blood cells was 1 in the detection assay. The testing result was not effected by red blood cells taken from either different red blood cell type of human or different individual of guinea pigs.

CONCLUSIONSThe novel method has several advantages: simple, high sensitivity and specificity, accurate and suitable for multiple purposes.

Animals ; Cells, Cultured ; Erythrocyte Aggregation ; drug effects ; Guinea Pigs ; Humans ; Influenza A virus ; enzymology ; isolation & purification ; Influenza B virus ; enzymology ; isolation & purification ; Influenza, Human ; diagnosis ; virology ; Neuraminidase ; analysis ; Reagent Kits, Diagnostic ; Sensitivity and Specificity

Due to the increasingly severe situation of pathogen resistance， the efficacy of classical antibiotics has declined， and the difficulty of treating community-acquired pneumonia （CAP） has gradually increased. Omadacycline was approved for marketing in China in December 2021 and has been approved for treating community-acquired bacterial pneumonia and acute bacterial skin and skin structure infection in adults. Based on introducing the pathogen resistance of CAP， this article systematically reviews the antibacterial activity and resistance， pharmacokinetic characteristics， and clinical efficacy of omadacycline. It is found that its antibacterial spectrum is wide， its bioavailability is high， and its safety is good. Its pharmacokinetic parameters are not affected by the patients’ age， gender， liver and kidney function， or drug interactions， and it is effective against multiple drug-resistant bacteria， Gram-negative bacteria producing extended-spectrum β-lactases， atypical pathogens， etc. It can be used as one of the treatment options for CAP. However， due to limited clinical research data and the short clinical application time of the drug， more clinical studies are still needed to verify its efficacy as a monotherapy or in combination with other antibiotics for severe pneumonia patients.

Objective:To investigate the changes in the nerve fiber layer of the cornea in patients with demyelinating optic neuritis (DON) and its correlation with visual acuity.Methods:A cross-sectional study. From March 2021 to July 2022, 27 cases (39 eyes) of DON patients diagnosed in the Department of Neurology and Ophthalmology of Beijing Tongren Hospital Affiliated to Capital Medical University were enrolled in this study. According to the serological test results, the patients were divided into aquaporin 4 antibody associated optic neuritis (AQP4-ON group) and myelin oligodendrocyte glycoprotein antibody associated optic neuritis (MOG-ON group), with 15 cases (19 eyes) and 12 cases (20 eyes) respectively. According to previous history of glucocorticoid treatment, the patients were divided into glucocorticoid treated group and non-glucocorticoid treated group, with 17 cases (27 eyes) and 10 cases (12 eyes) respectively. Twenty healthy volunteers (20 eyes) with age- and gender-matched were selected as the control group. All eyes underwent best corrected visual acuity (BCVA) and in vivo confocal microscopy (IVCM) examinations. BCVA was performed using Snellen's standard logarithmic visual acuity chart, which was converted into logarithmic minimum angle resolution (logMAR) visual acuity during statistics. The corneal nerve fiber length (CNFL), corneal nerve fiber density (CNFD), corneal nerve fiber branch length (CNBL), corneal nerve fiber branch density (CNBD) and the density of corneal dendritic cells (DC) were detected by IVCM examination. Parameter comparison between groups by t-test and Kruskal-Wallis rank sum test. The correlation between logMAR BCVA and pamameters of corneal nerve fibers were analyzed using Spearman analysis. Results:The CNFL, CNFD, and CNBL of the DON group and the control group were (10.67±2.55) mm/mm 2, (57.78±12.35) root/mm 2, (3.27±1.34) mm/mm 2, and (13.74±3.05) mm/mm 2, (70.95±13.14) root/mm 2, and (4.22±1.03) mm/mm 2, respectively; the difference in CNFL, CNFD, and CNBL between the two groups were statistically significant ( t=4.089, 3.795, 2.773; P<0.05). The CNFL, CNBL, and CNBD of the affected eyes in the MOG-ON group and AQP4-ON group were (12.02±2.13) mm/mm 2, (3.80±1.19) mm/mm 2, (47.97±8.86) fibers/mm 2, and (9.25±2.19) mm/mm 2, (2.72±1.19) mm/mm 2, (39.43±13.86) fibers/mm 2, respectively; the differences in CNFL, CNBL, and CNBD between the two groups were statistically significant ( t=-4.002, -2.706, -2.306; P<0.05). The corneal DC density of the patients in the hormone treated group and the non-hormone treated group was (24.43±8.32) and (41.22±9.86) cells/mm 2, respectively. The difference in corneal DC density between the two subgroups was statistically significant ( P<0.001). Correlation analysis showed that there was a significant negative correlation between logMAR BCVA and CNBL and CNFL in patients with DON ( r=-0.422, -0.456; P<0.05). Conclusions:There are different degrees of corneal nerve fiber damage in patients with different types of DON. There was a negative correlation between BCVA and the length of corneal nerve fibers.

Objective:To study the clinical characteristics of purulent meningitis complicated with hydrocephalus in neonates, and to analyze the risk factors of the disease.Methods:Neonates diagnosed with purulent meningitis complicated with hydrocephalus who hospitalized in the department of neonatology of the Second Affiliated Hospital of Wenzhou Medical University from January 2002 to August 2021 were selected as the case group. Neonates with positive pathogen cultures but no hydrocephalus during the same period were assigned by random number table method as the control group. The ratio of the control group and the case group was 2 ∶1. The clinical data such as bacteria distribution, cranial imaging, therapy and prognosis were compared between the two groups. The risk factors for hydrocephalus were predicted. Statistical analysis was conducted using chi-square test and multiple logistic regression analysis.Results:There were 33 cases in the case group and 66 cases in the control group. A total of 27 cases had confirmed pathogen results, of which 20 cases (74.1%) were Gram-negative bacteria and seven cases (25.9%) were Gram-positive bacteria. The time of diagnosis for hydrocephalus were 13.0(5.5, 28.5) days after the onset. Twenty-six cases received non-surgical treatment, while seven cases received surgery. The cure rate of case group was 42.4%(14/33), which was lower than that of control group (72.7%, 48/66), and the difference was statistically significant ( χ2=8.63, P=0.003). Univariate analysis showed that the incidences of protein>3 g/L in cerebrospinal fluid, glucose<2 mmol/L in cerebrospinal fluid, convulsions, central respiratory failure, intracranial hemorrhage and encephalomalacia in the case group were all higher than those in the control group, with statistical significance ( χ2=19.72, 12.04, 19.04, 5.73, 11.85 and 17.48, respectively, all P<0.050). Multivariate logistic regression analysis showed that convulsions (odds ratio ( OR)=4.476, 95% confidence interval ( CI) 1.091 to 18.363, P=0.037), intracranial hemorrhage ( OR=8.031, 95% CI 1.894 to 34.059, P=0.005) and encephalomalacia ( OR=35.189, 95% CI 2.954 to 419.150, P=0.005) were risk factors for neonatal purulent meningitis complicated with hydrocephalus. Conclusions:Gram-negative bacteria are common pathogen of neonatal purulent meningitis complicated with hydrocephalus. Convulsions, intracranial hemorrhage and encephalomalacia are important predictors for neonatal purulent meningitis complicated with hydrocephalus.

Objective:To identify the core genes related to the disease severity of respiratory syncytial virus (RSV) bronchiolitis in children using RNA sequencing (RNA-seq) and weighted gene co-expression network analysis (WGCNA), aiming to provide reference for predicting the condition of RSV infection.Methods:Twenty-two patients admitted to the Second Affiliated Hospital of Wenzhou Medical University with RSV bronchiolitis from October 1, 2019 to February 29, 2020 were enrolled as the case group. They were divided into three groups based on the severity of the disease: mild group, moderate group and severe group. Twenty-two healthy children were selected as the control group. Total RNA was extracted from whole blood leukocytes and analyzed by RNA-seq to compare the differentially expressed genes (DEGs) between children with RSV bronchiolitis and healthy children. The gene co-expression modules related to disease severity and biological indicators for disease severity assessment were identified.Results:The median age of the 22 patients (19 males and 3 females) was 3 months. The median age of the 22 healthy children (14 males and 8 females) was 4 months. There was no significant difference in age or gender between the two groups. There were 8 cases in the mild group, 7 cases in the moderate group and 7 cases in the severe group. Through significance analysis, 416 DEGs were found in the mild group, 586 in the moderate group and 846 in the severe group. According to WGCNA analysis, 10 co-expression modules were found, among which brown module ( r=0.62, P<0.001) was significantly correlated with disease severity. The protein-protein interaction network of DEGs in brown module was constructed and the top 30 core genes were selected according to the connectivity of gene nodes, among which the genes with high correlation were RBX1 and PSMA7. The expression of RBX1 and PSMA7 genes was up-regulated in the severe group, but their expression in the mild and moderate groups was not significantly different from that in the control group. Conclusions:RBX1 and PSMA7 genes might be biological predictors of disease severity in RSV bronchiolitis.

Objective:To evaluate the distribution and drug sensitivity of pathogenic bacteria in patients with bacterial keratitis in recent 40 years in China.Methods:A Meta-analysis was conducted.Five databases including PubMed, ScienceDirect, Embase, CNKI and Wanfang Data Knowledge Service Platform were searched.The research was limited to the hospital-based cross-sectional studies published in Chinese and English between 1980 and 2020, without limitations of subject.Two researchers followed the inclusion and exclusion criteria to complete literature retrieval, data extraction and methodological quality evaluation.The literature quality was assessed with reference to a methodological scoring system for rates. Q test and I2 test were used to quantify the degree of heterogeneity of the included literature.According to the heterogeneity, the fixed effects model or random-effects model was used to calculate the combined rates indicators to perform the Meta-analysis. Results:Twenty-seven original studies were incorporated, including 50 046 cases of bacterial keratitis.A random-effects model for the Meta-analysis showed that the positive rate of bacterial culture in bacterial keratitis was 28% (95% CI: 0.24-0.32). Among the culture-positive bacteria, the percentages of gram-positive cocci and gram-negative bacilli were 57% (95% CI: 0.52-0.62) and 32% (95% CI: 0.28-0.37), respectively.The percentages of gram-positive bacilli and gram-negative cocci were 8% (95% CI: 0.06-0.10) and 1% (95% CI: 0.01-0.02), respectively.In the recent 40 years, the proportion of gram-positive cocci isolated from corneas in China was on the rise, and the proportion of gram-negative bacilli was on the decline.Among them, coagulase-negative Staphylococcus accounted for 23% (95% CI: 0.17-0.30), followed by Pseudomonas aeruginosa 18% (95% CI: 0.14-0.23), Streptococcus pneumoniae 8% (95% CI: 0.06-0.12), Staphylococcus aureus 6% (95% CI: 0.04-0.08), Corynebacterium 4% (95% CI: 0.03-0.07), and Escherichia coli 4% (95% CI: 0.02-0.06). The results of the drug sensitivity test showed that gram-positive cocci were highly sensitive to vancomycin, moxifloxacin, levofloxacin and ofloxacin. Pseudomonas aeruginosa was most sensitive to tobramycin, and Escherichia coli was most sensitive to ofloxacin. Conclusions:This Meta-analysis indicates that the proportion of gram-positive cocci isolated from corneas in China has been on the rise, while the proportion of gram-negative bacilli was on the decline during the recent 40 years.Coagulase-negative Staphylococcus and Pseudomonas aeruginosa have become the most common pathogens of bacterial keratitis in China.The sensitivity of various bacteria to commonly used antibiotics shows a downward trend.The sensitive antibiotics should be selected correctly according to the drug sensitivity results.

Diabetic retinopathy(DR)is one of the common causes of visual impairment and blindness in adults, which is caused by various pathogenesis. Although the mechanism of DR has not been elucidated yet, the destruction of blood-retinal barrier is a key process. As a highly endothelial-specific factor in promoting the growth of vascular endothelial cell, vascular endothelial growth factor(VEGF)plays a crucial role in the formation of pathological retinal neovascularization and the destruction of blood-retinal barrier. Therefore, a comprehensive understanding of the etiology and pathogenesis of blood-retinal barrier damage promoted by VEGF is critical for exploring the pathogenesis of DR. In this study, the underlying relationship between VEGF and the mechanism of blood-retinal barrier damage, including retinal vascular endothelial cell permeability, vascular inflammatory response, apoptosis, oxidative stress, mitochondrial damage and endoplasmic reticulum stress, with a view to providing a reference for the study in VEGF in the pathogenesis of blood-retinal barrier damage in DR.

cFos is one of the most widely-studied genes in the field of neuroscience. Currently, there is no systematic database focusing on cFos in neuroscience. We developed a curated database-cFos-ANAB-a cFos-based web tool for exploring activated neurons and associated behaviors in rats and mice, comprising 398 brain nuclei and sub-nuclei, and five associated behaviors: pain, fear, feeding, aggression, and sexual behavior. Direct relationships among behaviors and nuclei (even cell types) under specific stimulating conditions were constructed based on cFos expression profiles extracted from original publications. Moreover, overlapping nuclei and sub-nuclei with potentially complex functions among different associated behaviors were emphasized, leading to results serving as important clues to the development of valid hypotheses for exploring as yet unknown circuits. Using the analysis function of cFos-ANAB, multi-layered pictures of networks and their relationships can quickly be explored depending on users' purposes. These features provide a useful tool and good reference for early exploration in neuroscience. The cFos-ANAB database is available at www.cfos-db.net .
Animals ; Fear ; Mice ; Neurons ; Proto-Oncogene Proteins c-fos ; Rats