Objective Currently, there is no commercially available quantitative detection kit for the main Felis domestic allergen (Fel d 1) in China. To establish a rapid detection method for Fel d 1, this study aims to prepare monoclonal antibodies against Fel d 1 protein. Methods The codon preference of Escherichia coli was utilized to optimize and synthesize the Fel d 1 gene. The prokaryotic expression plasmid pET-28a-Fel d 1 was constructed and used to express and purify the recombinant Fel d 1 protein. Subsequently, the recombinant protein was immunized into BALB/c mice and monoclonal antibodies (mAbs) were prepared by the hybridoma technique. An indirect ELISA was established using the recombinant Fel d 1 as the coating antigen, and hybridoma cell lines were screened for positive clones. The specificity and antigenic epitopes of the mAbs were confirmed by Western blot analysis. Finally, the selected hybridoma cells were injected into the peritoneal cavities of BALB/c mice for large-scale monoclonal antibody production. Results The recombinant plasmid pET-28a-Fel d 1 was successfully constructed, and soluble Fel d 1 protein was obtained after optimizing the expression conditions. Western blot and antibody titer assays confirmed the successful isolation of two hybridoma cell lines, 7D11 and 5H4, which stably secreted mAbs specific to Fel d 1. Antibody characterization revealed that the 5H4 mAb was of the IgG2a subtype and could recognize the amino acid region 105-163 of Fel d 1, while the 7D11 mAb was the IgG1 subtype and could recognize the amino acid region 1-59. Conclusion The high-purity recombinant Fel d 1 protein produced in this study provides a promising alternative for clinical immunotherapy of cat allergies. Furthermore, the monoclonal antibody prepared in this experiment lays a material foundation for the in-depth study of the biological function of Fel d 1 and the development of ELISA detection.
Animals ; Antibodies, Monoclonal/biosynthesis* ; Mice, Inbred BALB C ; Cats ; Mice ; Allergens/genetics* ; Glycoproteins/genetics* ; Enzyme-Linked Immunosorbent Assay ; Hybridomas/immunology* ; Recombinant Proteins/genetics* ; Female ; Antibody Specificity

Objective: To reveal the molecular mechanism underlying the compatibility of Salvia miltiorrhiza Bge (S. miltiorrhiza, Dan Shen) and C. tinctorius L. (C. tinctorius, Hong Hua) as an herb pair through network pharmacology and subsequent experimental validation. Methods: Network pharmacology was applied to construct an active ingredient-efficacy target-disease protein network to reveal the unique regulation pattern of S. miltiorrhiza and C. tinctorius as herb pair. Molecular docking was used to verify the binding of the components of these herbs and their potential targets. An H9c2 glucose hypoxia model was used to evaluate the efficacy of the components and their synergistic effects, which were evaluated using the combination index. Western blot was performed to detect the protein expression of these targets. Results: Network pharmacology analysis revealed 5 pathways and 8 core targets of S. miltiorrhiza and C. tinctorius in myocardial protection. Five of the core targets were enriched in the hypoxia-inducible factor-1 (HIF-1) signaling pathway. S. miltiorrhiza-C. tinctorius achieved vascular tone mainly by regulating the target genes of the HIF-1 pathway. As an upstream gene of the HIF-1 pathway, STAT3 can be activated by the active ingredients cryptotanshinone (Ctan), salvianolic acid B (Sal. B), and myricetin (Myric). Cell experiments revealed that Myric, Sal. B, and Ctan also exhibited synergistic myocardial protective activity. Molecular docking verified the strong binding of Myric, Sal. B, and Ctan to STAT3. Western blot further showed that the active ingredients synergistically upregulated the protein expression of STAT3. Conclusion The pharmacodynamic transmission analysis revealed that the active ingredients of S. miltiorrhiza and C. tinctorius can synergistically resist ischemia through various targets and pathways. This study provides a methodological reference for interpreting traditional Chinese medicine compatibility.

Irritable bowel syndrome (IBS) is a functional gastrointestinal disorder that is characterized by abdominal pain and disordered bowel habits. The etiology of IBS is multifactorial, including abnormal gut-brain interactions, visceral hypersensitivity, altered colon motility, and psychological factors. Recent studies have shown that the intestinal microbiota and its metabolites short chain fatty acids (SCFAs) may be involved in the pathogenesis of IBS. SCFAs play an important role in the pathophysiology of IBS. We discuss the underlying mechanisms of action of SCFAs in intestinal inflammation and immunity, intestinal barrier integrity, motility, and the microbiota-gutbrain axis. Limited to previous studies, further studies are required to investigate the mechanisms of action of SCFAs in IBS and provide more precise therapeutic strategies for IBS.

Objective:To evaluate the characteristics of carotid plaque and the immediate outcomes after carotid artery stenting (CAS) in diabetic and non-diabetic patients by optical coherence tomography (OCT).Methods:Patients underwent CAS and OCT before and after operation in the Department of Neurology, Jinling Hospital from January 2014 to March 2019 were enrolled retrospectively. The clinical features, the characteristics of carotid plaque on OCT and the immediate outcomes after CAS were compared between diabetic group and non-diabetic group. The risk factors of stent malapposition were analyzed.Results:A total of 46 patients were enrolled. Their age was 64.02±8.32 years and 41 were males (89.1%). There were 20 patients (43.5%) in the diabetes group and 26 (56.5%) in the non-diabetes group. The proportions of atherosclerotic plaque with thin fibrous cap (40.0% vs. 7.7%; χ2=5.166, P=0.023), plaque rupture (55.0% vs. 23.1%; χ2=4.945, P=0.026) and macrophage infiltration (60.0% vs. 30.8%; χ2=3.930, P=0.047) in the diabetic group were significantly higher than those in the non-diabetic group. Multivariate logistic regression analysis showed that older age (odds ratio [ OR] 1.208, 95% confidence interval [ CI] 1.033-1.413; P=0.018), coronary heart disease ( OR 15.953, 95% CI 1.142-222.952; P=0.040), alcohol consumption ( OR 6.192, 95% CI 1.098-34.923; P=0.039) and lower systolic blood pressure ( OR 0.944, 95% CI 0.894-0.997; P=0.037) were independently associated with stent malaposition. Conclusion:Compared with the non-diabetic patients, carotid plaque in diabetic patients may be more unstable. Older age, coronary heart disease, alcohol consumption and lower systolic blood pressure were associated with stent malaposition after carotid stenting. OCT can reveal the characteristics of carotid plaque and the immediate outcomes after CAS, which can provide strong evidence for treatment decision.

Objective:To understand the epidemiological characteristics of infectious diarrhea pathogens in Pudong New Areas of Shanghai from 2013 to 2017 to provide evidence for control and prevention of the disease.Methods:From Jan 2013 to Dec 2017, active surveillance program on diarrhea was conducted in 14 sentinel hospitals (three tertiary-level and nine secondary-level, and two primary-level hospitals) in Pudong New Areas of Shanghai, based on location, catchment areas and number of patients. All recruited outpatients were interviewed in hospitals, using a standard questionnaire. Stool specimens were collected and tested for five viral and eight bacterial pathogens.Results:A total of 9 301 cases with infectious diarrhea were included, and the overall positive rate was 55.7 % (5 179). Positive rates of single virus, single bacteria and mixed infections were 26.7 % (2 481), 17.0 % (1 579) and 12.0 % (1 119), respectively. For single infection, the most commonly detected viruses appeared as norovirus (15.4 %, 1 428/9 301) and rotavirus (7.2 %, 667/9 301). The most commonly detected bacteria were diarrheagenic Escherichia coli (6.7 %, 619/9 301) and non-typhoid Salmonella (3.3 %, 305/9 301). The most common mixed infections were caused by virus-bacteria (4.9 %, 459/9 301). Norovirus (17.0 %, 838/4 938) showed the highest positive rates, followed by Escherichia coli (7.2 %, 354/4 938), both seen in the age group of 20-59 years old group. Rotavirus (9.4 %, 178/1 896) and non-typhoid Salmonella (4.9 %, 93/1 896) were the most common pathogens found in the age group of 0-4 years old. The prevalence of norovirus peaked both in spring and autumn. The other peaks were seen as: Rotavirus in winter, diarrheagenic Escherichia coli in summer and non-typhoid Salmonella in summer. Conclusions:Our data showed that the positive rates of infectious diarrhea pathogens were high in Pudong New Areas of Shanghai from 2013 to 2017. The dominant pathogens would include norovirus, rotavirus and diarrheagenic Escherichia coli but with differenct distributions in age groups. Obvious seasonal patterns were also observed.

Objective: To investigate the antimicrobial resistance and molecular epidemiology of foodborne Yersinia (Y.) enterocolitica in Pudong New District of Shanghai. Methods: Four kinds of raw food samples were collected in retail circulation sites in Pudong from 2012 to 2016. Cold enrichment method was used to isolate Y. enterocolitica and further detection of biotype, serotype, virulent genes, antimicrobial susceptibility of the isolates and pulsed field gel electrophoresis (PFGE) were conducted. Results: A total of 3 900 raw food samples were collected during this period, including poultry product (n=590), livestock product (n=1 074), aquatic product (n=1 488), vegetable (n=748), in which 111 (2.8%) were contaminated by Y. enterocolitica. The detection rates of Y. enterocolitica in poultry product samples (5.3%, 31/590) and livestock product samples (4.5%, 48/1 074) were higher than those in aquatic product samples (1.6%, 24/1 488) and vegetable samples (1.1%, 8/748). The predominant biotype was 1A (95.5%) and predominant serotype was O∶8 (42.3%). All the strains lacked ail, ystA, yadA and virF genes, which encoded pathogenic Y. enterocolitica. Seventy six (68.5%) strains harbored ystB gene, in which 35 (31.5%) belonged to 1A/O∶8/ystB pattern. Most strains were resistant to ampicillin (74.8%) and amoxicillin/clavulanic acid (70.3%), and non-sensitive rate to Cefoxitin was over 50.0%. No third generation cephalosporin or fluoroquinolone resistant strains were detected, but 38.7% (43/111) strains were multidrug resistant (MDR). Serotype O∶8 and O∶5 strains had 44 and 18 PFGE patterns, respectively. Conclusions The main foodborne exposure sources of Y. enterocolitica in raw food were poultry and livestock products in Pudong New District. 1A/O∶8/ystB was the predominant pattern with potential pathogenicity despite lacks of typical pathogenic virulent genes. The antimicrobial resistant rates of Y. enterocolitica were at a low level, but MDR strains still existed. Molecular types of the isolates showed highly genetic diversity.

Stroke is the first cause of death in China. Intravenous thrombolysis and early endovascular treatment can significantly improve the outcome of patients with ischemic stroke. With the continuous deepening of research, the time window of endovascular interventional therapy has been extended from 6 h to 24 h. Tissue stratification plays a vital role in the expansion of time window. This article reviews the implications of tissue stratification, imaging assessment methods, and their significance in endovascular treatment.

Objective: To study the clinical characteristics of children with adenovirus pneumonia and provide evidence for clinical diagnosis and treatment timely. Method: This retrospective study included 89 children who were confirmed to have adenovirus pneumonia in hospital from January 2015 to December 2016. All the immunofluorescence test result of the 89 children showed that the exfoliated nasopharyngeal cells from the 89 children were all adenovirus antigen positive. All the severe type children reached the diagnostic criteria of severe pneumonia by the respiratory group in the society of pediatrics, Chinese Medical Association. The children were divided into 2 groups (severe type group and common type group). Different factors such as epidemiologic feature, clinical manifestation, laboratory examination and imaging data were analyzed. Results: Among the 89 pediatric patients, the male to female ratio was 1.5∶1. The ages ranged from 1 month to 14 years. Children under 5 years of age accounted for 96.6%(86/89). The incidence was 37.1%(33/89)in winter and 30.3%(27/89)in spring. The lengths of hospital stay were 3-48 days and the median length of stay was 8.25±4.75 days. All of these 89 cases had fever and cough. The proportion of severe adenovirus pneumonia was high among male, under 2 years of age, those with dyspnea, hepatosplenomegaly, tachycardia, leukocytosis, elevated C-reactive protein (CRP), PCT, myocardial enzymes, electrocardiogram abnormality and cluster shadow in chest CT. Differences were statistically significant (P<0.05). Conclusions Special attention should be paid to the male children under the 2 years of age with abnormal related indicators to consider severe adenovirus pneumonia. Early detection and treatment are the key to improve treatment and reduce death.

Objective To discuss the alterations of functional brain properties in patients with idiopathic trigeminal neuralgia (ITN) based on graph theoretical approaches.Methods A total of 20 patients with pain in the distribution of left trigeminal nerve and 22 matched healthy controls were recruited in our hospital from May 2013 to January 2015.Resting-state functional MRI data in both groups were collected and analyzed with GRETNA software to obtain the matrices of brain network.From these connectivity matrices,a number of key characteristics were computed,including network efficiency,weighted shortest path length,weighted clustering coefficient and small-wordless.A two-tailed t test was used to appraise the significance of any inter-group differences in complex network measures investigated.Network-based statistic (NBS) approach was used to localize specific pairs of brain regions in which functional connectivity strength were altered in patients.Pearson's correlation analyses were performed to identify the correlations between clinical data and altered network properties in patients.Results ITN patients had significantly increased weighted clustering coefficient in the right calcarine (CAL) and bilateral thalamus,increased total efficiency in the left superior frontal gyrus (SFG),and increased regional efficiency in the right SFG,right CAL,bilateral cuneus and left pallidum,and significantly reduced weighted shortest path length for regional nodal characteristics in the right superior parietalis lobules (P＜0.05).The total efficiency in the left SFG and regional efficiency in the right SFG in ITN patients were negatively correlated with course of disease (r=-0.500,P=0.035;r=-0.501,P=0.034).As compared with the healthy controls,the ITN patients had enhanced functional loop connection between left SFG and right supplementary motor area,between left SFG and right SFG,between left SFG and right middle frontal gyrus,between left SFG and right orbital middle frontal gyrus,between left SFG and partes opercularis of right gyrus frontalis inferior (IFGoperc),between left SFG and bilateral putamen (PUT),between left SFG and right PAL,between right PUT and left interior superior frontal gyrus,and between left PUT and right caudate nucleus.The connectivity strength between left SFG and right IFGoperc was negatively correlated with disease duration (r=-0.487,P=-0.041).Conclusion Alterations in regional nodal characteristics may suggest central sensitization and modulation in ITN patients.

Objective To identify the diffusion alterations of deep gray matter(GM) and white matter (WM) among Alzheimer's disease (AD), mild cognitive impairment (MCI) and healthy people by atlas?based analysis (ABA), and to investigate the respective relationship with cognitive function. Methods Twenty?one AD patients (AD group), 8 MCI patients (MCI group) and 15 normal controls (control group) were performed by conventional MRI and diffusion tensor imaging (DTI). The raw data of DTI was processed by using DTI studio software to generate the fractional anisotropy (FA) images. Then ABA was used to quantify the FA value in 58 deep GM and WM structures. The differences of FA value among three groups were compared by using one way ANOVA, with a post?hoc analysis. In AD and MCI groups, the partial correlation was further investigated between mini?mental state examination (MMSE) score and FA value in the brain regions that have significant differences between AD and MCI group or between MCI and control group. Results Compared with control group, AD patients showed wide?spread FA decrease in most deep GM and WM regions (corrected P<0.05). The FA values of the hypothalamus, the fornix, the superior longitudinal fasciculus (SLF) and the cingulum in AD group were significantly lower than those in MCI group (corrected P<0.05). The FA value of the right splenium of corpus callosum (SCC) in MCI group was significantly lower than that in control group (MCI:0.550±0.018 vs. Control:0.585±0.026, P<0.05). In AD and MCI group, the FA values of the left hypothalamus, the right hypothalamus, the left cingulum, the right cingulum, and the left SLF were positively correlated with MMSE scores(r=0.502, 0.515, 0.535, 0.527, 0.512; P<0.05). No significant correlation was found between the FA value of the right SCC, the right SLF, the right fornix/stria terminalis, the right fornix and MMSE scores(P>0.05). Conclusion Based on ABA, this study found the diffusion changes not only in the WM but also the deep GM in AD patients, but only WM diffusion disruptions in MCI group. The decreased FA value in the right SCC appeared early, but had no correlation with the cognitive impairment. The FA value in the hypothalamus, the fornix, the SLF and the cingulum decreased with the disease progression, and correlated positively with the cognition decline.