Cross-reaction mediated by distinct key amino acid combinations in the complementary-determining region (CDR) of a monoclonal antibody.

In immunology, cross-reaction between antigens and antibodies are commonly observed. Prior research has shown that various monoclonal antibodies (mAbs) can recognize a broad spectrum of epitopes related to influenza viruses. However, existing theories on cross-reactions fall short in explaining the phenomena observed. This study explored the interaction characteristics of H1-74 mAb with three peptides: two natural peptides, LVLWGIHHP and LPFQNI, derived from the hemagglutinin (HA) antigen of the H1N1 influenza virus, and one synthetic peptide, WPFQNY. Our findings indicate that the complementarity-determining region (CDR) of H1-74 mAb comprised five antigen-binding sites, containing eight key amino acid residues from the light chain variable region and 16 from the heavy chain variable region. These critical residues formed distinct hydrophobic or hydrophilic clusters and functional groups within the binding sites, facilitating interaction with antigen epitopes through hydrogen bonding, salt bridge formation, and π-π stacking. The study revealed that the formation of the antibody molecule led to the creation of binding groups and small units in the CDR, allowing the antibody to attach to a variety of antigen epitopes through diverse combinations of these small units and functional groups. This unique ability of the antibody to bind with antigen epitopes provides a new molecular basis for explaining the phenomenon of antibody cross-reaction.

Fibronectin binds integrin α5ß1 to regulate macular neovascularization through the Wnt/ß-catenin signaling pathway.

Age-related macular degeneration (AMD) is a progressive, degenerative disease of the macula. The formation of macular neovascularization (MNV) and subretinal fibrosis of AMD is the most classic cause of the loss of vision in older adults worldwide. While the underlying causes of MNV and subretinal fibrosis remain elusive, the common feature of many common retinal diseases is changes the proportions of protein deposition in extracellular matrix (ECM) when compared to normal tissue. In ECM, fibronectin (FN) is a crucial component and plays a pivotal part not only in fibrotic diseases but also in the process of angiogenesis. The study aims to understand the role of ligand FN and its common integrin receptor α5ß1 on MNV, and to understand the molecular mechanism involved. To study this, the laser-induced MNV mouse model and the rhesus macaque choroid-retinal endothelial cell line (RF/6A) chemical hypoxia mode were established, and the FN-α5ß1 expression levels were detected by immunohistochemistry (IHC) and quantitative real-time PCR analysis (qRT-PCR). Fibronectin expression was silenced using small interfering RNA (siRNA) targeting FN. The tube formation and vitro scratch assays were used to assess the ability to form blood vessels and cell migration. To measure the formation of MNV, immunofluorescence, and Western blot assays were used. These results revealed that the expressions of FN and integrin α5ß1 were distinctly increased in the laser-induced MNV mouse model and in the RF/6A cytochemically induced hypoxia model, and the expression tendency was identical. After the use of FN siRNA, the tube formation and migration abilities of the RF/6A cells were lower, the ability of endothelial cells to proliferate was confined and the scope of damage caused by the laser in animal models was significantly cut down. In addition, FN gene knockdown dramatically inhibited the expression of Wnt/ß-catenin signal. The interaction of FN with the integrin receptor α5ß1 in the constructed model, which may act through the Wnt/ß-catenin signaling pathway, was confirmed in this study. In conclusion, FN may be a potential new molecular target for the prevention and treatment of subretinal fibrosis and MNV.

Effects of congruent emotional contexts during encoding on recognition: An ERPs study.

Past research showed that emotional contexts can impair recognition memory for the target item. Given that item-context congruity may enhance recognition memory, the present study aims to examine the effect of the congruent emotional encoding contexts on recognition memory. Participants studied congruent word-picture pairs (e.g., the word "cow" - a picture describing a cow) and incongruent word-picture pairs (e.g., the word "cow" - a picture describing a goat) and, subsequently, were asked to report the nature of the picture (emotional or neutral). Behavioral results revealed that emotional contexts impaired source but not item recognition, with congruent word-context mitigating this impairment and enhancing item recognition. Neural results from ERPs and theta oscillations found the recollection process, as shown by the LPC old/new effect and theta oscillations, for both item and source recognition across emotional contexts, irrespective of congruity. Meanwhile, the familiarity process as indexed by the FN400 old/new effect was found only for item recognition in congruent emotional contexts. These findings suggest that the congruent relationship of item-context could mitigate the emotion-induced source memory impairment and enhance item memory, with neural results elucidating the memory processes involved in retrieval of emotional information. Specifically, while emotion-related information generally elicits the recollection-based memory process, only congruent emotional information elicits the familiarity-based process.

Improving memory through choice and deliberation in decision-making: Evidence from ERPs.

The goal of decision-making is to select one option and disregard the others. However, deliberation can also create a memory association between the chosen and unchosen options. This study aims to investigate how choice and deliberation affect the memory of postdecision options and the underlying mechanisms. Using event-related potentials (ERPs), we examined item recognition (Experiment 1) and associative recognition (Experiment 2) following certain and uncertain decisions. In Experiment 1, items that were chosen in certain decisions were remembered better than unchosen items. There was no difference between chosen and unchosen items in uncertain decisions. Moreover, a late recollection-related LPC (a late positive component) old/new effect was larger for chosen items than unchosen items in certain decisions. The early familiarity-related FN400 and the late recollection-related LPC old/new effects were significant for chosen and unchosen items in uncertain decisions. In Experiment 2, there was no difference in performance on associative memory. A FN400 old/new effect (an index of integration) in certain or uncertain decisions was not observed. Although significant LPC old/new effects were found in both certain and uncertain decisions, no difference was found between them. These results propose that decision-making can enhance item memory performance through two distinct processes: value and elaboration. Elaboration involves focusing on the details within items rather than integrating items into a cohesive whole.

Simultaneous determination of 18 steroids in the hypothalamic pituitary gonadal axis based on UPLC-MS/MS with multimode ionization.

To objectively quantify changes in steroid hormones in organisms caused by adverse environmental loads, we developed a simple and sensitive UPLC-MS/MS (ultra-performance liquid chromatography triple quadrupole mass spectrometry) method for the simultaneous determination of 18 steroid hormones on the HPG axis. This analytical method was based on liquid extraction and a multimode electrospray and atmospheric pressure chemical ionization (ESCi) source, which was optimized by mass spectrometry, liquid phase and pretreatment for the quantification of cholesterol (CH), aldosterone (A), cortisone (E), hydrocortisone (F), 21-deoxycortisol (21-DF), corticosterone (B), 11-deoxycortisol (11-DF), androstenedione (A2), estradiol (E2), estrone (E1), 2-methoxyestradiol (2-MeE2), 21-hydroxyprogesterone (21-OHP), 17-α hydroxyprogesterone (17α-OHP), testosterone (T), dehydroepiandrosterone (DHEA), progesterone (P4), dihydrotestosterone (DHT), and pregnenolone (P5). The method exhibits linearity in the analyte-concentration range 0.03-1000 µg mL-1 (r2 > 0.99), the spiked recoveries for the concentration range tested are 76.22-113.66%, and the relevant parameters of precision are 7.52-1.14%. Compared to other methods, this new method not only uses a small amount of serum (only 100 µL), but also permits the analysis of the challenging steroid, cholesterol. Furthermore, the method was successfully applied to the determination of steroids in Mus musculus, Carassius auratus, Rana catesbeiana Shaw, and Rana nigromaculata serum samples from randomly selected individuals. Therefore, this method is efficient and a very useful tool for assessing changes in steroid hormones.

Identification of Heterophilic Epitopes of H1N1 Influenza Virus Hemagglutinin.

Our previous studies found that the H1-50 monoclonal antibody (mAb) of influenza A virus hemagglutinin (HA) cross-reacted with pancreatic tissue and islet ß-cells, and further studies showed that H1-50 mAb binds to prohibitin (PHB) protein of islet ß-cells. These suggest that there are heterophilic epitopes between influenza virus HA and pancreatic tissue, which may be involved in the pathogenesis of type 1 diabetes. To further investigate these heterophilic epitopes, we screened binding epitopes of H1-50 mAb using a phage 12-peptide library. DNA sequencing and comparative analysis were performed on specific positive phage clones, and the sequence of 12-peptide binding to H1-50 mAb was obtained. The binding epitopes of H1-50 mAb in influenza virus HA were determined by sequence analysis and experimental verification, and their distribution within the three-dimensional structure was assessed by PyMOL. The results showed that H1-50 mAb specifically binds to polypeptides (306-SLPFQNIHPITIGK-319) of influenza A virus HA, located in the stem of the HA protein. However, there is no specific binding sequence between H1-50 mAb and the PHB protein of islet ß-cells in the primary structure, and we speculate that the binding of H1-50 mAb to islet ß-cells may depend on the spatial conformation. The identification of the heterophilic epitopes of H1N1 influenza virus hemagglutinin provides a new perspective on type 1 diabetes that may be caused by influenza virus infection, which may contribute to the prevention and control of influenza.

Simulation analysis and physiological and biochemical evaluation of Sophora flavescens aboveground against aphids using network pharmacology.

Pests cause substantial damage to human environments; therefore, studying insecticidal mechanisms is crucial for improving pest control. However, the use of chemical pesticides can cause irreversible secondary damage. In this study, we used network pharmacology to investigate the effect of Sophora flavescens Alt., as a biological pest control agent, on glucose-6-phosphate 1-dehydrogenase, thymidylate synthase, and a translocation protein in aphids. The stability and reliability of target proteins was analyzed using molecular docking and molecular dynamic simulations. Enzyme activity assays validated the feasibility of network pharmacology to obtain actionable targets. We used interdisciplinary integration to study pest control and network pharmacology to identify how Sophora flavescens Alt. resists aphid attacks. The results show that the use of network pharmacology can increase the accuracy and specificity of our predictions for the molecules targeted by insecticides. This approach will facilitate improved, environmentally friendly pest control development in the future.

Identification and analysis of B cell epitopes of hemagglutinin of H1N1 influenza virus.

The frequent variation of influenza virus hemagglutinin (HA) antigen is the main cause of influenza pandemic. Therefore, the study of B cell epitopes of HA is of great significance in the prevention and control of influenza virus. In this study, the split vaccine of 2009 H1N1 influenza virus was used as immunogen, and the monoclonal antibodies (mAbs) were prepared by conventional hybridoma fusion and screening techniques. The characteristics of mAbs were identified by ELISA method, Western-blot test and hemagglutination inhibition test (HI). Using the obtained mAbs as a tool, the B cell epitopes of HA were predicted by ELISA blocking test, sandwich ELISA method and computer simulation method. Finally, four mAbs against HA antigen of H1N1 influenza virus were obtained. The results of ELISA and computer prediction showed that there were at least two types of epitopes on HA of influenza virus. The results of this study complemented the existing methods for predicting HA epitopes, and also provided a new method for predicting other pathogenic microorganisms.

First-Line Immunochemotherapy Versus Palliative Chemotherapy Plus Definitive Radiation Therapy for de novo Metastatic Nasopharyngeal Carcinoma: A Matched Cohort Study.

BACKGROUND: The combined use of immune checkpoint inhibitors (ICIs) with palliative chemotherapy (PCT) is a promising first-line treatment for de novo metastatic nasopharyngeal carcinoma (mNPC). However, the efficacy of ICIs with PCT vs PCT with definitive radiation therapy (DRT) remain unclear. METHODS: Patients with mNPC who received first-line immunochemotherapy (ICI + PCT) or PCT + DRT were included. Propensity score matching (PSM) was applied to balance potential confounders between patients who did and did not undergo DRT (at a ratio of 1:1). Progression free survival (PFS) and overall survival (OS) were compared between the 2 groups using a log-rank test and Cox proportional hazard model. RESULTS: Among all participants, 149 received ICI + PCT. After PSM, 149 patients were included in the PCT + DRT group. First-line immunochemotherapy was associated with significantly improved PFS (median 9.0 months vs 12.0 months, P < .001) and OS (median 12.5 months vs 19.9 months, P < .001). Subgroup analysis revealed that tumor response to immunochemotherapy, metastatic organs, and number of metastatic sites potentially affected the efficacy of DRT after first-line immunochemotherapy. CONCLUSION: Compared with PCT + DRT, first-line immunochemotherapy was associated with improved PFS and OS in patients with mNPC but not in patients with unfavorable tumor response and metastasis involving the liver, distant nodes, or multiple sites.

A natural cuttlefish melanin nanoprobe for preoperative and intraoperative mapping of lymph nodes.

Lymphatic metastasis plays an important role in malignant tumor invasion. Efficient identification of sentinel lymph node (SLN) is extremely significant for designing therapeutic strategies and assessing prognosis. In this work, we developed a natural cuttlefish melanin nanoprobe for the preoperative and intraoperative evaluation of lymphatic metastasis. The cuttlefish melanin nanoparticle could improve the water-solubility and biocompatibility of the near-infrared-II (NIR-II) dye, and extend the retention time of small molecule dye. The NIR-II imaging results verified that the nanoparticles have a high accumulation, high sensitivity, and high signal-to-noise ratio in the lymphatic system. Moreover, the nanoparticles have obvious naked-eye identification potential due to their natural brownish-black color. Additionally, the nanoparticles can combine with Gd ions to achieve preoperative lymphatic magnetic resonance imaging (MRI). The results of this study provide a unique approach to effectively identify and accurately remove lymph nodes before operation and during surgery, exhibiting tremendous potential in clinical translation.

First Report of Alternaria alternata Inciting Leaf Spot of Dangshen (Codonopsis pilosula) in China.

As a commonly traditional Chinese medicine, the perennial herb Dangshen (Codonopsis pilosula) has superior curative effects including regulating immunity, strengthening the spleen, and tonifying lungs (Bai et al. 2020). To imitate natural ecological conditions, plants were grown on hillside fields with stems prostrate on the ground, tangle-up with each other. In August 2020, leaf spots were observed on C. pilosula in Wutai county, Shanxi province, China, and indicated a high disease incidence (70%-80%) in investigated fields (6.67 ha). Small brown necrotic spots, occasionally enclosed by chlorotic halos, were observed on leaves, stems, and sepals. For identification of the pathogen, 15 small pieces (5×5 mm) of symptomatic tissues from 5 randomly-collected diseased plants were surface sterilized, placed on potato dextrose agar plates, and incubated for 4 d in darkness at 25 °C to obtain the colonies. Cultures were purified by single spore isolation from these colonies. A total of 15 isolates named as Dcp-3, and Dcp-5~Dcp-18 were recovered. They produced ovoid or obclavate spores with 15.9-57.5×9.1-20.1 µm in size, 1-6 transverse septa, and 0-4 longitudinal septa. The conidial chains with 4 to 6 spores had numerous secondary and occasionally tertiary chains on potato carrot agar plates. Because all isolates had identical morphological traits, five genes from the representative isolate Dcp-3, actin (ACT), Alternaria major allergen (Alt a1), plasma membrane ATPase (ATP), histone 3 (H3), and rDNA ITS, were amplified with primer pairs ACTDF1/ACTDR1, Alt-for/Alt-rev, ATPDF1/ATPDR1, H3-1a/H3-1b, and ITS1/ITS4, respectively (Hong et al. 2005; Lawrence et al. 2013; Ma et al. 2020). BLASTn searches indicated species of Dcp-3 could not be accurately confirmed by rDNA ITS, ATP, ACT, and Alt a1 (GenBank accession nos. OM334894, OM362504, OM326344, OM362500). Phylogenetic analysis showed it was most closely related to Alternaria alternata, A. arborescens, and A. tenuissima based on concatenated sequences of above four genes. The H3 sequence (OM362508) shared 100% homology with that of A. alternata (MN481948). The phylogenetic tree using H3 also confirmed Dcp-3 as A. alternata. Heathly, two-year-old C. pilosula were transplanted to a greenhouse. A surface-sterilized leaf was sprayed with 50 µL spore suspension (106 conidia/mL) of Dcp-3. A leaf sprayed with isometric sterile water was used as controls. Each treatment used six plants (five leaves per plant). Plants were covered with sterilized plastic bags and incubated at 22 ℃. The test was repeated twice. A week later, control leaves were healthy, but brown necrotic spots similar to field symptoms emerged on treated leaves. The A. alternata isolates were re-isolated from the border of lesions, and confirmed by morphological and molecular characteristics mentioned above, fulfilling Koch's postulates. Leaf spot of C. pilosula caused by Septoria codonopsidis has been reported in China (Wang et al. 2011). However, to our knowledge, this is the first report of A. alternata inciting leaf spot of C. pilosula in China. Our report would promote growers to enhance the field management and consider associated strategies on controlling Alternaria leaf spot of C. pilosula.

Preparation, Characterization, and Evaluation of Liposomes Containing Oridonin from Rabdosia rubescens.

Due to the remarkable anti-tumor activities of oridonin (Ori), research on Rabdosia rubescens has attracted more and more attention in the pharmaceutical field. The purpose of this study was to extract Ori from R. rubescens by ultrasound-assisted extraction (UAE) and prepare Ori liposomes as a novel delivery system to improve the bioavailability and biocompatibility. Response surface methodology (RSM), namely Box-Behnken design (BBD), was applied to optimize extraction conditions, formulation, and preparation process. The results demonstrated that the optimal extraction conditions were an ethanol concentration of 75.9%, an extraction time of 35.7 min, and a solid/liquid ratio of 1:32.6. Under these optimal conditions, the extraction yield of Ori was 4.23 mg/g, which was well matched with the predicted value (4.28 mg/g). The optimal preparation conditions of Ori liposomes by RSM, with an ultrasonic time of 41.1 min, a soybean phospholipids/drug ratio of 9.6 g/g, and a water bath temperature of 53.4 °C, had higher encapsulation efficiency (84.1%). The characterization studies indicated that Ori liposomes had well-dispersible spherical shapes and uniform sizes with a particle size of 137.7 nm, a polydispersity index (PDI) of 0.216, and zeta potential of -24.0 mV. In addition, Ori liposomes presented better activity than free Ori. Therefore, the results indicated that Ori liposomes could enhance the bioactivity of Ori, being proposed as a promising vehicle for drug delivery.

Neutralizing antibody PR8-23 targets the footprint of the sialoglycan receptor binding site of H1N1 hemagglutinin.

Influenza virus cause seasonal influenza epidemic and seriously sporadic influenza pandemic outbreaks. Hemagglutinin (HA) is an important target in the therapeutic treatment and diagnostic detection of the influenza virus. Variation in the sialic acid receptor binding site leads to strain-specific binding and results in different binding modes to the host receptors. Here, we evaluated the neutralizing activity and hemagglutination inhibition activity of a prepared murine anti-H1N1 monoclonal antibody PR8-23. Then we identified the epitope peptide of antibody PR8-23 by phage display technique from phage display peptide libraries. The identified epitope, 63-IAPLQLGKCNIA-74, containing two α-helix and two ß-fold located at the footprint of the sialoglycan receptor on the RBS in the globular head domain of HA. It broads the growing arsenal of motifs for the amino acids on the globular head domain of HA in sialic acid receptor binding site and neutralizing antibody production.

Improvement of long-term clinical outcomes by successful PCI in the very elderly women with ACS.

BACKGROUND: Whether very elderly women with acute coronary syndromes (ACS) should receive aggressive percutaneous coronary intervention (PCI) is still controversial. We assessed the effectiveness and long-term clinical outcomes of successful PCI in this population and identified prognostic factors which might contribute to the incidence of major adverse cardiovascular and cerebrovascular events (MACCE) in the very elderly female PCI cohort. METHODS: Female ACS patients aged ≥ 80 years were consecutively enrolled (n = 729) into the study. All the patients were divided into female PCI group (n = 232) and medical group (n = 497). MACCE was followed up, including non-fatal myocardial infarction (MI), stroke, heart failure requiring hospitalization (HFRH), cardiovascular (CV) death, and the composite of them. After propensity score matching (1:1), the incidences of MACCE were compared between the two groups. Clinical and coronary artery lesion characteristics were compared between the female PCI patients with (n = 56) and without MACCE (n = 176). Multivariate Cox regression analysis was performed to identify risk factors which independently associated with MACCE in the female PCI patients. MACCE of male PCI patients, who aged ≥ 80 years and hospitalized in the same period (n = 264), was also compared with that of the female PCI patients. RESULTS: A total of 32% very elderly female ACS patients received PCI in the present study. (1) Compared to female medical group, PCI procedure significantly alleviated the risks of MACCE: non-fatal MI (6.2% vs. 20.2%, P < 0.001), HFRH (10.9% vs. 22.5%, P = 0.012), CV death (12.4% vs. 28.7%, P < 0.001) and the composite MACCE (24.0% vs. 44.2%, P < 0.001) during the median follow-up period of 36 months. (2) Between very elderly female and male PCI patients, there were no significant differences in occurrence of MACCE (P = 0.232) and CV death (P = 0.951). (3) Multivariate Cox analysis revealed that ST-segment elevation myocardial infarction (STEMI) (HR 1.944, 95% CI 1.11-3.403, P = 0.02) and elevated log- N-Terminal pro-brain natriuretic peptide (NT-proBNP) (HR 1.689, 95% CI 1.029-2.773, P = 0.038) were independently associated with the incidence of MACCE in the female PCI patients. CONCLUSIONS: PCI procedure significantly attenuated the risk of MACCE and improved the long-term clinical outcomes in very elderly female ACS patients. Aggressive PCI strategy may be reasonable in this population.

Monoclonal Antibody Targeting the HA191/199 Region of H1N1 Influenza Virus Mediates the Damage of Neural Cells.

Vaccination is the most effective mean of preventing influenza virus infections. However, vaccination-induced adverse reactions of the nervous system, the causes of which are unknown, lead to concerns on the safety of influenza A vaccine. In this study, we used flow cytometry, cell ELISA, and immunofluorescence to find that H1-84 monoclonal antibody (mAb) against the191/199 region of the H1N1 influenza virus hemagglutinin (HA) protein binds to neural cells and mediates cell damage. Using molecular simulation software, such as PyMOL and PDB viewer, we demonstrated that the HA191/199 region maintains the overall structure of the HA head. Since the HA191/199 region cannot be removed from the HA structure, it has to be altered via introducing point mutations by site-directed mutagenesis. This will provide an innovative theoretical support for the subsequent modification the influenza A vaccine for increasing its safety.

Unitization could facilitate item recognition but inhibit verbatim recognition for picture stimuli: behavioral and event-related potential study.

It is widely accepted that unitization can promote familiarity-based associative recognition, but the effect of unitization on item recognition remains unclear. The goals of this study were to elaborate on how unitization affected item recognition and the neural correlates of familiarity and recollection for item recognition. In study, the participants were asked to learn related and unrelated picture pairs, and in test, they were required to distinguish single old pictures form new pictures. In experiment 1, we used R/K/N paradigm to estimate the contribution of familiarity and recollection to item recognition, the results showed that unitization could improve item recognition through increasing recollection selectively. In experiment 2, we used ERP old/new effects to estimate the neural correlates of familiarity and recollection, the results showed that unitization could improve item recognition through a selective reduction in LPC effect. Inspired by DRM paradigm, in experiment 3, we divided the new pictures into semantically related lure pictures and semantically unrelated new pictures to explore the effects of unitization on item recognition and verbatim recognition (the ability to distinguish old pictures from lure pictures). The behavioral results showed that unitization could improve item recognition, but it damaged verbatim recognition. The ERP results revealed that there were larger LPC effects in the unrelated condition than in the related condition, regardless of item or verbatim recognition. In summary, we believed that unitization could indeed improve item recognition, but it damaged verbatim recognition.

The effect of unitization on associative recognition was not moderated by the unitization-congruence between original and rearranged picture pairs (UC) for picture stimuli.

It is widely accepted that associative recognition can be supported by familiarity through integrating more than two stimuli into a unit, but the role of unitization in recollection-based associative recognition remains unclear. In this study, we aimed to illustrate how the level of unitization (LOU) affected recollection-based associative recognition and to examine whether the unitization-congruence between original and rearranged picture pairs (UC) could have effect on the relationship between LOU and associative recognition. In encoding, participants were asked to learn related and unrelated picture pairs, and in retrieval, they needed to distinguish intact pairs from rearranged pairs. We also distinguished the LOU of the pairs based on its status at encoding or retrieval separately. The results showed that: (1) LOU-at-encoding could improve associative recognition through increasing recollection-based associative recognition selectively; (2) LOU-at-retrieval could improve associative recognition through increasing familiarity-based and recollection-based associative recognitions; (3) UC did not moderate the relationship between LOU and associative recognition. Hence, in future studies, researchers do not need to pay much attention to the construction of rearranged pairs to ensure that the LOU between original and rearranged picture pairs is matched. It greatly reduces the difficulty of materials selection.

The treatment of macrolide-resistant Mycoplasma pneumoniae pneumonia in children.

WHAT IS KNOWN AND OBJECTIVE: In recent years, the resistance of Mycoplasma pneumoniae to macrolide antibiotics has increased significantly. The health systems are facing significant challenges in carrying out the diagnosis and treatment of refractory Mycoplasma pneumoniae pneumonia in children. Levofloxacin is suitable for treating infectious diseases in various systems but limited in children due to arthropathy issues in weight-bearing joints. This study aimed to evaluate the efficacy and safety of levofloxacin in children with macrolide-resistant Mycoplasma pneumoniae pneumonia. METHODS: We retrospectively enrolled six confirmed cases of refractory Mycoplasma pneumoniae pneumonia who were admitted in the paediatric respiratory ward of Shandong provincial hospital Affiliated to Shandong first Medical University between 1st January 2020 and 29th February 2020. Levofloxacin was given to the patients through the intravenous or oral route as per the following dosages :<5 years, 8-10 mg/kg q12 h; >5 years, 8-10 mg/kg, qd for 10 days. The clinical data were collected and analysed. RESULTS AND DISCUSSION: The average age of the enrolled cases was six years and nine months (range, four years, and seven months to eleven years and seven months). All cases were found to be drug-resistant and were treated with azithromycin combined with antibacterial drugs. Levofloxacin was used in the patient's refractory to macrolide antibiotics. The temperature of five cases returned to normal 1-2 days after treatment with levofloxacin, and the imaging of the four cases showed expected improvements. The gastrointestinal symptoms, neurological manifestations, joint symptoms, blood parameters, liver and kidney functions, and exercise conditions of the children were closely monitored. The follow-up time of the patients ranged from one week to five months. No drug-related adverse reactions were observed in patients during treatment or during follow-up. WHAT IS NEW AND CONCLUSION: The clinical symptoms and imaging significantly improved after treatment with levofloxacin, and no drug-related adverse reactions were observed. Levofloxacin proved to be an effective and safe drug in the treatment of children with macrolide-resistant mycoplasma pneumonia. This study will provide a reference for evaluating the efficacy and safety of levofloxacin in the paediatric population.

Under the condition of unitization at encoding rather than unitization at retrieval, familiarity could support associative recognition and the relationship between unitization and recollection was moderated by unitization-congruence.

It is widely accepted that associative recognition can be supported by familiarity through integrating more than two stimuli into a unit, but there are still three unsolved questions: (1) how unitization affects recollection-based associative recognition; (2) whether it is necessary to match the level of unitization (LOU) between original and rearranged pairs, which was term as unitization-congruence (UC); (3) whether unitization can occur at encoding or at retrieval. The purposes of this study are to try to answer these questions. During the encoding phase, the participants were asked to learn compound words and unrelated word pairs, and during the retrieval phase, they needed to distinguish intact pairs from rearranged consistent and rearranged inconsistent pairs with "remember/know" paradigm. The results showed that (1) the role of unitization in recollection was moderated by UC; (2) Under the consistent UC condition, unitization could improve familiarity-based associative recognition without affecting recollection-based associative recognition, while under the inconsistent UC condition, unitization could improve familiarity-based and recollection-based associative recognition simultaneously, these results indicated that it was necessary to match the LOU between original and rearranged pairs; (3) unitization at encoding could support familiarity-based associative recognition, while unitization at retrieval did not. In briefly, unitization at encoding could improve associative recognition and this effect was moderated by UC, while unitization at retrieval did not affect associative recognition.

[Evidence of clinical randomized controlled trial study in treatment of acute cerebral infarction with traditional Chinese medicine in recent five years].

To systematically search and sort out the clinical randomized controlled trial(RCT) on the prevention and treatment of acute cerebral infarction with traditional Chinese medicine(TCM) by using the method of evidence map, and to understand the evidence distribution of related studies. CNKI, Wanfang, VIP, CBM, PubMed, EMbase, Cochrane Library and Web of Science were retrieved from January 2016 to September 2020, and literatures related to the prevention and treatment of acute cerebral infarction with traditional Chinese medicine were included. Text description combined with table and bubble chart were used to analyze the distribution characteristics of evidence. A total of 1 102 clinical articles in recent five years were retrieved. The annual trend of clinical study publication, study size, TCM therapy category and main scheme, and study literature quality were analyzed. We find that TCM treatment of acute cerebral infarction has become a hot topic of clinical research, the number of literature showed a trend of increased year by year, various means of intervention of TCM in the treatment of the advantages of increasingly highlight. Follow-up clinical research should highlight the characteristics of TCM: in the analysis of outcome indicators; increase the neuropsychological patients after stroke and cognitive ability, and the theory of combined treatment of TCM disease when thoughts; At the same time, the quality of clinical research needs to be improved. At present, there is still a lack of unified standards for the production of evidence map. This study is the first to explore the application of evidence map to summarize and display the clinical research status of TCM treatment of acute cerebral infarction, and combine it with the setting of priority areas of TCM clinical research, so as to provide a reference basis for determining the priority topic selection of TCM treatment optimization research.