Therapeutic Characterization of ^(131)I-Labeled Humanized Anti-B7-H3 Antibodies for Radioimmunotherapy for Glioblastoma

B7 homolog 3(B7-H3)has attracted much attention in glioblastoma(GBM)radioimmunotherapy(RIT)due to its abnormally high expression on tumor cells.In this study,we report that two specific humanized anti-human B7-H3 antibodies(hu4G4 and hu4H12)derived from mouse anti-human B7-H3 antibodies that were generated by computer-aided design and exclusively recognize membrane expression of B7-H3 by human glioma cells,Hu4G4 and hu4H12 were radiolabeled with^(89)Zr for RIT antibody screening.Micro-positron emission tomography(PET)imaging,biodistribution and pharmacokinetic(PK)analyses of^(89)Zr-labeled antibodies were performed in U87-xenografted models.^(125)I labelling of the antibodies for single-photon emission computed tomography(SPECT)imaging was also used to investigate the biological behavior of the antibodies in vivo.Fu rthermore,the pharmacodynamic(PD)of the^(131)Ilabeled antibodies were evaluated in U87-xenografted mice and GL261 Red-FLuc-B7-H3 in situ glioma tumor models.Micro-PET imaging and biodistribution analysis with a gamma counter showed that^(89)Zr-deferoxamine(DFO)-hu4G4 had higher tumor targeting performance with lower liver uptake than^(89)Zr-DFO-(hu4H12,immunoglobulin G(IgG)).The biodistribution results of^(125)I-SPECT imaging were similar to those of^(89)Zr-PET imaging,though the biodistribution in long bone joints and the thyroid varied.The PD analysis results indicated that^(131)I-hu4G4 had an excellent therapeutic effect and high safety with no apparent toxicity.Interestingly,^(131)I-hu4G4 improved the tumor vasculature in tissues with higher expression of collagen typeⅣand platelet-derived growth factor receptorβ(PDGFR-β)compared with control treatment,as determined by immunofluorescence(IF),which contributed to inhibiting tumor growth.Taken together,our data indicate that hu4G4 exhibits good tumor targeting and specificity,achieves low nonspecific concentrations in normal tissues,and has acceptable PK characteristics.^(131)I-hu4G4 also exerts effective antitumor effects with an ideal safety profile.Therefore,we expect hu4G4 to be an excellent antibody for the development of GBM RIT.

LncRNA HIF1A-AS2 accelerates malignant phenotypes of renal carcinoma by modulating miR-30a-5p/SOX4 axis as a ceRNA

Objective:Several reports have proposed that lnc RNAs,as potential biomarkers,participate in the progression and growth of malignant tumors.HIF1 A-AS2 is a novel lnc RNA and potential biomarker,involved in the genesis and development of carcinomas.However,the molecular mechanism of HIF1 A-AS2 in renal carcinoma is unclear.Methods:The relative expression levels of HIF1 A-AS2 and miR-30 a-5 p were detected using RT-qPCR in renal carcinoma tissues and cell lines.Using loss-of-function and overexpression,the biological effects of HIF1 A-AS2 and miR-30 a-5 p in kidney carcinoma progression were characterized.Dual luciferase reporter gene analysis and Western blot were used to detect the potential mechanism of HIF1 A-AS2 in renal carcinomas.Results:HIF1 A-AS2 was upregulated in kidney carcinoma tissues when compared with para-carcinoma tissues(P<0.05).In addition,tumor size,tumor node mestastasis stage and differentiation were identified as being closely associated with HIF1 A-AS2 expression(P<0.05).Knockdown or overexpression of HIF1 A-AS2 either restrained or promoted the malignant phenotype and WNT/β-catenin signaling in renal carcinoma cells(P<0.05).Mi R-30 a-5 p was downregulated in renal cancers and partially reversed HIF1 A-AS2 functions in malignant renal tumor cells.HIF1 A-AS2 acted as a micro RNA sponge that actively regulated the relative expression of SOX4 in sponging miR-30 a-5 p and subsequently increased the malignant phenotypes of renal carcinomas.HIF1 A-AS2 showed a carcinogenic effect and miR-30 a-5 p acted as an antagonist of the anti-oncogene effects in the pathogenesis of renal carcinomas.Conclusions:The HIF1 A-AS2-miR-30 a-5 p-SOX4 axis was associated with the malignant progression and development of renal carcinoma.The relative expression of HIF1 A-AS2 was negatively correlated with the expression of miR-30 a-5 p,and was closely correlated with SOX4 mRNA levels in renal cancers.

EXPRESSION OF DPC4 PROTEIN IN COLORECTAL CARCINOMA IN DIFFERENT STAGES

Objective.To d etermine whether the non-expression of DPC4protein only occurs late in the dev el-opment of colorectal carcinoma.Methods.In this study,we examined the ex pression of DPC4protein in formalin-fixed archival specimens from102colorec tal neoplasm with immunohistochemical analysis.Those specimens were classi-fie d into5stages:stageⅠ;stageⅡ(intramucosal carcinoma ,8cases);stageⅢ(primary invasive carcinoma without infiltration of the l ymph nodes,11cases);stageⅣ(primary invasive carcinoma with infiltration of the lymph nodes,25cases);and stageⅤ(carcinoma metastasized to dis-ta nt tissue,22cases).Results.The frequency of non-expression of DPC4prote ins were5.5%in stage I,12.5%in stage II;9% in stage III;36%in stage IV;32%in stag e V.The frequency of negative expression of DPC4protein were analyzed by÷ 2 test for stage II and III versus stage IV and V and there was statistically significant difference.At same time ,there was statistically signifi-cant differencefor adenoma versus carcinoma .Conclusions.The frequency of non-expression of DPC4protein increases as the stage of colorectal carcinoma advances and the non-expressio n of DPC4protein is likely to be a late event in the sequential pathogenesis o f colorectal carcinoma.The non-expression DPC4protein in colorectal neoplasm may sug-gest its malignant characteristic,which will help us to increase the insight on colorectal carcinoma.

Functional Conjugation of the Different Regulatory Responses to the Stress Stimuli in Healthy Human Subjects

Present article discusses the physiological mechanisms of the state employees adaptation during active training in temporary groups. It is suggested that adaptive mechanisms to adverse effects may be studied basing on the concept of functional isomorphism of the psychic and immune systems. Adaptive mechanisms were studied through the monitoring of the stress factors’ impact upon the law enforcement officers when training outside the places of permanent deployment. The specific purpose of present study was to evaluate the physiological indicators of the psychic, immune and endocrine systems dynamics at different stages of adaptation of the live organism to a stressful situation, hoping to get better insight into possible relations between psychic and immune domains. Through monitoring of the dynamics of the endocrine and immune responses to the psychic stimuli, it was possible to correlate the stages of the stress onset to the phases of specific immune reactions. Strong correlations between the parameters characterizing activation of the psychic and immune responses support the hypothesis of the presence of “strong cooperation” between psychic and immune domains. It supports earlier hypothesis that we are monitoring different manifestations of the functioning of a complex inseparable psycho-immune meta-system.

Investigating the Mechanism of Qu Du Qiang Fei 1 Hao Fang Formula against Coronavirus Disease 2019 Based on Network Pharmacology Method

Objective:Qu Du Qiang Fei 1 Hao Fang(QDQF1)is a novel Chinese herbal medicine formula used to treat coronavirus disease 2019(COVID-19).However,the pharmacological mechanisms of action of QDQF1 remain unclear.The objective of this study was to identify the effective ingredients and biological targets of QDQF1 for COVID-19 treatment.Materials and Methods:The effective ingredients and mechanisms of action of QDQF1 were analyzed by using network pharmacology methods,which included an analysis of the effective ingredients and corresponding targets,COVID-19-related target acquisition,compound-target network analyses,protein-protein interaction network analysis,Kyoto Encyclopedia of Genes and Genomes(KEGG)and Gene Ontology(GO)enrichment analyses,and molecular docking studies.Results:In total,288 effective QDQF1 ingredients were identified.We identified 51 core targets from the 148 targets through an overlap between putative QDQF1 targets and COVID-19-related targets.Six key components,including formononetin,kaempferol,luteolin,naringenin,quercetin,and wogonin were identified through component-target network analyses.GO functional enrichment analysis of the core targets revealed 1296 items,while KEGG pathway enrichment analysis identified 148 signaling pathways.Nine central targets(CCL2,CXCL8,IL1B,IL6,MAPK1,MAPK3,MAPK8,STAT3,and TNF)related to the COVID-19 pathway were identified in the KEGG pathway enrichment analysis.Furthermore,molecular docking analysis suggested that the docking scores of the six key components to the nine central targets were better than those to remdesivir.Conclusions:QDQF1 may regulate multiple immune-and inflammation-related targets to inhibit the progression of severe acute respiratory syndrome coronavirus 2,and thus,may be suitable for the treatment of COVID-19.

Clinical Study with Randomized Control on the Therapy of Integrated Chinese and Western Medicine in Treating Neurological Autoimmune Diseases:A Meta-Analysis

Objective: The main objective of this study is to evaluate the effects of integrated Chinese and Western medicine in the treatment of neurological autoimmune diseases. Materials and Methods: The literature was comprehensively searched to collect the randomized controlled trials about integrated Chinese and Western medicine in the treatment of neurological autoimmune diseases. Neurological autoimmune diseases mainly occur in the central nervous system(CNS) and peripheral nervous system. Therefore, multiple sclerosis(MS) was chosen as the representative in the CNS, and Guillain–Barre syndrome(GBS) was chosen as the representative in the peripheral nervous system. Extended Disability Status Scale(EDSS) score, effective rate, clinical symptom score, neurological functional sign score, recurrence frequency, and incidence rate of adverse reactions were chosen as the markers of outcome variables of MS, and the effective rate and Hughes score were also chosen as the markers of outcome variables of GBS. Results: For MS, the results showed that there was a significant difference in statistical analysis between the experimental group and the control group in EDSS score, the effective rate, and the recurrence frequency. However, through the comparison of clinical symptom score, neurological functional sign score, and incidence rate of adverse reaction of both two groups, the results showed that there was no significant difference in the statistical analysis. For GBS, through the comparison of effective rate and Hughes score of both two groups, the results showed that there was a significant difference in statistical analysis. Conclusions: The study demonstrated that compared with Western medicine, the therapy of integrated Chinese and Western medicine was more effective in treating neurological autoimmune diseases.

PD-L1 is increased in the spinal cord and infiltrating lymphocytes in experimental allergic encephalomyelitis

Experimental allergic encephalomyelitis is a mouse model of human multiple sclerosis with similar pathology and pathogenesis. Thl cells play an important role in the pathogenesis of experimental allergic encephalomyelitis. This study determined the potential effect of programmed cell death 1 ligand 1 in the pathogenesis of experimental allergic encephalomyelitis induced by injecting myelin oligodendrocyte glycoprotein, complete Freund＇s adjuvant and Bordetella pertussis toxin into C57BL/6J mice. Experimental allergic encephalomyelitis mice developed disease and showed in- flammatory changes in the central nervous system by hematoxylin-eosin staining of spinal cord pathological sections, demyelination by Luxol fast-blue staining and clinical manifestations. The expression of programmed cell death 1 ligand 1 in mice was detected by immunohistochemistry, flow cytometry and western blot anatysis. The expression of programmed cell death 1 ligand 1 in the spinal cord and splenocytes of mice was significantly increased compared with normal mice. Our findings suggest the involvement of programmed cell death 1 ligand 1 in the pathogenesis of ex- perimental allergic encephalomyelitis and suggest this should be studied in multiple sclerosis.

Antibody Therapies Targeting Complex Membrane Proteins

In analyses of protein families that may serve as drug targets,membrane-associated G-protein-coupled receptors(GPCRs)dominate,followed by ion channels,transporters,and—to a lesser extent—membrane-bound enzymes.However,various challenges put such membrane proteins among key groups of underutilized opportunities for the application of therapeutic antibodies.Antibodies hold the promise of exquisite specificity,as they are able to target even specific conformations of a particular membrane protein,as well as adaptability through engineering into various antibody formats.However,the ease of raising and isolating specific,effective antibodies targeting membrane proteins depends on many factors.In particular,the generation of specific antibodies is easier when targeting larger,simpler,extracellular domains with greater uniqueness of amino acid sequence.The rareness of such ideal conditions is illustrated by the limited number of approved biologics for targeting GPCRs and other complex membrane proteins.Challenges in developing antibodies to complex membrane proteins such as GPCRs,ion channels,transporters,and membrane-bound enzymes can be addressed by the design of the antigen,antibody-generation strategies,lead optimization technologies,and antibody modalities.A better understanding of the membrane proteins being targeted would facilitate mechanism-based drug discovery.This review describes the advantages and challenges of targeting complex membrane proteins with antibodies and discusses the preparation of membrane protein antigens and antibody generation,illustrated by select examples of success.

Tau protein,phosphorylated tau protein,and beta-amyloid 42 levels in patients with neurodegenerative diseases complicated by cognitive deficits A non-randomized,concurrent,case-control investigation

BACKGROUND： The differential diagnosis of many neurodegenerative disorders depends primarily on clinical symptoms together with imaging methods. Recently, increased importance has been placed on the use of biomarkers for diagnosing various neurodegenerative disorders. OBJECTIVE： To assess the feasibility of tau-protein, phosphorylated tau-protein, beta-amyloid 42 （Aβ42）, and 14-3-3 protein as biomarkers for diagnosing several neurodegenerative diseases complicated by cognitive deficits. DESIGN, TIME AND SETTING： A non-randomized, concurrent, case-control investigation was performed in three medical centers in the Czech Republic （Department of Neurology at the University Hospital in Hradec Kralove, Department of Neurology at the 2rd Medical Faculty, and the University Hospital Motol） between October 2000 and November 2006. PARTICIPANTS： Eighteen patients with probable AIzheimer＇s disease, 4 patients with Creutzfeldt-Jakob disease, 10 patients with frontotemporal dementia, 9 patients with clinically isolated syndrome suggestive of multiple sclerosis, and 7 patients with multiple sclerosis, as well as 38 race-, nationality-, and age-matched cognitively intact controls, were included in the study. Diagnoses were established based on the following criteria： the criteria for Alzheimer＇s disease proposed by the National Institute of Neurological and Communicative Disorders and Stroke/Alzheimer＇s Disease and Related Disorders Association, WHO criteria for Creutzfeldt-Jakob disease, Neary criteria for frontotemporal dementia, and McDonald＇s criteria for multiple sclerosis. All included patients were confirmed to suffer from various degrees of dementia. METHODS： Enzyme-linked immunosorbent assay was used to measure concentrations of tau-protein, phosphorylated tau-protein, and Aβ42 in cerebrospinal fluid （CSF） samples collected by standard lumbar puncture from each patient. Moreover, 14-3-3 protein was assessed by Western blot in CSF of Creutzfeldt-Jakob disease patients. Cognitive status was assessed using the Mini Mental Scale Examination （MMSE） in all subjects. MAIN OUTCOME MEASURES： Establishment of biomarkers with greatest specificity and sensitivity for the investigated disorders according to Receiver Operating Characteristic curves, which were based on values from patients and controls; correlation between concentrations of given biomarkers and demographic parameters, diagnosis, duration of disease, and level of cognitive deficit. RESULTS： Increased concentrations of total tau protein and phosphorylated tau protein, and decreased levels of Aβ42, in CSF of Alzheimer＇s disease patients reached the required sensitivity/specificity ratio of 80% or greater. A marked elevation in CSF concentrations of total tau protein showed even greater sensitivity than 14-3-3 protein in Creutzfeldt-Jakob disease. There was no association between selected biomarkers and frontotemporal dementia or multiple sclerosis. Phosphorylated tau-protein was the only biomarker that noticeably correlated with MMSE scores for Alzheimer＇s disease.CONCLUSION： Levels of total tau protein, phosphorylated tau protein, and A!342 in the CSF could differentiate patients with Alzheimer＇s disease and Creutzfeldt-Jakob disease from healthy controls and patients with other neurodegenerative disorders. The diversity of absolute values demonstrates the necessity to establish a specific standard for each laboratory.

Tuberculosis Lymphadenitis in Northwest Ethiopia: Implications for Public Health

Objective: In Ethiopia where there are limited diagnostic facilities, the actual burden of tuberculosis (TB) lymphadenitis is not well known. Therefore, we conducted this study to determine the proportion of TB lymphadenitis (TBL) in childhood and adults in Northwest Ethiopia. Materials and Methods: A prospective cross sectional study was conducted from April to May 2012. Fine needle aspiration cytology (FNAC) for cytological diagnosis of TBL was used. The diagnosis of TBL was established when cytological features from lymph node aspirates are strongly suggestive of TB. Descriptive and multivariate analysis was done using SPSS version 16. Results: Out of 1070 patients attending the cytological diagnosis in the study sites 437 (41%) were positive for TBL. Of the 437 registered TBL, 59 (13.5%) were pediatric patients and 378 (86.5%) were adults. There were more females than males with a male to female ratio of 0.8:1. The cervical region had the most common group of TBL with 321 (73.2%) patients. Most of all these patients 314 (75.3%), were matted with a majority of (250/314) being in the cervical region. Individuals who had contact history with TB patients (P = 0.046) were more likely to have TBL. Conclusions: In the studied region high prevalence of TBL was documented. Screening of TBL particularly for those who had contact with TB patients is recommended. Most patients in our study presented with matted lymph nodes, indicating that late arrival of patients to health institutions. Thus continuous and intensified public health strategies on health education and early referral system have to be done in order to link them to health institutions earlier than at present.

Development of PREDAC-H1pdm to model the antigenic evolution of influenza A/(H1N1)pdm09 viruses

The Influenza A(H1N1)pdm09 virus caused a global pandemic in 2009 and has circulated seasonally ever since.As the continual genetic evolution of hemagglutinin in this virus leads to antigenic drift,rapid identification of antigenic variants and characterization of the antigenic evolution are needed.In this study,we developed PREDAC-H1pdm,a model to predict antigenic relationships between H1N1pdm viruses and identify antigenic clusters for post-2009 pandemic H1N1 strains.Our model performed well in predicting antigenic variants,which was helpful in influenza surveillance.By mapping the antigenic clusters for H1N1pdm,we found that substitutions on the Sa epitope were common for H1N1pdm,whereas for the former seasonal H1N1,substitutions on the Sb epitope were more common in antigenic evolution.Additionally,the localized epidemic pattern of H1N1pdm was more obvious than that of the former seasonal H1N1,which could make vaccine recommendation more sophisticated.Overall,the antigenic relationship prediction model we developed provides a rapid determination method for identifying antigenic variants,and the further analysis of evolutionary and epidemic characteristics can facilitate vaccine recommendations and influenza surveillance for H1N1pdm.

Therapeutic potential of traditional Chinese medicine for the treatment of NAFLD:A promising drug Potentilla discolor Bunge

Nonalcoholic fatty liver disease(NAFLD)is characterized by excessive accumulation of hepatic lipids and metabolic stress-induced liver injury.There are currently no approved effective pharmacological treatments for NAFLD.Traditional Chinese medicine(TCM)has been used for centuries to treat patients with chronic liver diseases without clear disease types and mechanisms.More recently,TCM has been shown to have unique advantages in the treatment of NAFLD.We performed a systematic review of the medical literature published over the last two decades and found that many TCM formulas have been reported to be beneficial for the treatment of metabolic dysfunctions,including Potentilla discolor Bunge(PDB).PDB has a variety of active compounds,including flavonoids,terpenoids,organic acids,steroids and tannins.Many compounds have been shown to exhibit a series of beneficial effects for the treatment of NAFLD,including anti-oxidative and anti-inflammatory functions,improvement of lipid metabolism and reversal of insulin resistance.In this review,we summarize potential therapeutic effects of TCM formulas for the treatment of NAFLD,focusing on the medicinal properties of natural active compounds from PDB and their underlying mechanisms.We point out that PDB can be classified as a novel candidate for the treatment and prevention of NAFLD.

Electroacupuncture Attenuates Surgical Stress-Induced Reduction of T Lymphocytes through Modulation of Peripheral Opioid System

Objective:To investigate the action mechanisms of electroacupuncture(EA)on postoperative immunosuppression.Methods:Male C57 BL/6 mice(5–7 weeks old)were randomly divided into:the sham injury group,the surgical trauma stressed group,the EA group[surgery+2/100 Hz EA at Neiguan(PC 6)],and the EA+Nal(surgery+EA+intraperitoneal injection of naloxone).Abdominal surgical trauma stress mice model was established.EA was performed on bilateral PC 6 acupoints by an EA apparatus(2/100 Hz)for 20 min once a day for 3 days.The m RNA expressions of MOR,DOR,and KOR in thymus and L3-5 dorsal root ganglions(DRG)were determined by quantitative real-time polymerase chain reaction(q RT-PCR)and the protein expressions of MOR,DOR,and KOR in thymus were measured by Western blot.Flow cytometry assay was used to detect the levels of T lymphocyte subtypes in the peripheral blood.Results:Surgical trauma induced decreased the m RNA expression level of MOR in both thymus(P<0.01)and L3-L5 DRGs(P<0.05).Moreover,EA treatment not only significantly attenuated the MOR protein and m RNA expression in the thymus(both P<0.05),but also markedly increased expression of DOR and KOR opioid receptor in thymus(P<0.01).However,the m RNA expressions of opioid receptors were not regulated by EA in the DRG(all P>0.05).Furthermore,T lymphocyte population of CD3^+ and CD4^+ was decreased in the peripheral blood after surgical trauma(both P<0.01).EA treatment can significantly elevate the population of CD3^+(P<0.01),CD4^+(P<0.05)and CD8^+T cells(P<0.01).Intraperitoneal injection of the non-selective opioid receptor antagonist naloxone blocked the up-regulation of T lymphocytes by EA.Conclusion:EA may improve postoperative immunosuppression through the peripheral opioid system.

Mycophenolic Acid Synergizing with Lipopolysaccharide to Induce Interleukin-1β Release via Activation of Caspase-1

Background： The previous study showed that mycophenolic acid （MPA） synergizing with lipopolysaccharide （LPS） promotcd interleukin （IL）-1β release, but the mechanism is unclear. This study aimed to investigate the nlechanism ofMPA synergizing with LPS to induce IL-1β release. Methods： Undiluted human blood cells, THP- I human rnyeloid leukemia mononuclear cells （THP- 1 ） cells, or monocytes were stimulated with L PS and treated with or without M PA, and the supernatant IL-1β was detected by enzyme-linked immunosorbent assay. The m RN A levels of IL- 1β were detected by real-time quantitative polymerase chain reaction. The intracellular protein levels of nuclear factor kappa B （NF-κ） phospho-p65 （p-p65）, precursor interleukin-1β （pro-IL-1β）, NOD-like receptor pyrin domain containing-3 （NLRP3）, and cysteine aspartic acid-specific protease-1 （caspase-1 ） p20 in THP-1 cell were measured by Western blot. Results： The MPA alone failed to induce IL-1β, whereas MPA synergized with LPS to increase IL-1β in a dose-dependent manner （685.00 ± 20.00 pg/ml in LPS ＋ 5 μmol/L MPA group, P =0.035; 742.00 ± 31.58 pg/ml in LPS ＋ 25 μmol/L MPA group, P = 0.017：1000.00 ± 65.59 pg/ml in LPS ＋ 75 μmol/L MPA group, P = 0.024： versus 408.00 ± 35.50 pg/ml in LPS group）. MPA alone has no effect on the IL-1β mRNA expression, LPS induced the expression of IL-1β mRNA 2761 fold, and LPS ＋ MPA increased the IL-1β expression 3018 fold, which had the same effect with LPS group （P = 0.834）. MPA did not affect the intracellular NF-κB p-p65 and pro-IL-1β protein levels but activated N LRP3 inflammasonae. Ac-YVAD-cmk blocked the activation ofcaspase-1 and subsequently attenuated IL- 1β secretion （ 181.00 ± 45.24 pg/ml in LPS ＋ M PA ＋ YVAD group vs. 588.00 ± 41.99 pg/ml in LPS ＋ M PA group, P= 0.014）. Conclusions： Taken together, MPA synergized with LPS to induce IL- 1β release via the activation of caspase- 1, rather than the enhanced production ofpro-IL-1β. These findings suggested that patients immunosuppressed with mycophenolate mofetil may have overly activated caspase- 1 during infection, which might contribute to a more sensitive host defense response to invading germs.

PREDAV-H1: a user-friendly web server for predicting antigenic variants of influenza H1N1 viruses

Dear Editor,The influenza H1N1 virus has caused three global pandemics since the beginning of the 20th century(Liu et al.,2015).The first is the notorious Spanish flu in 191&which killed 20-100 million people in the world.It circulated for nearly 40 years and was replaced by influenza H2N2 virus.In 1977,the virus reappeared in Russia and caused global pandemics.It continued to circulate until 2009 when it was replaced by the pandemic H1N1 virus.Influenza H1N1 viruses cause large morbidity and mortality to human society,and will continue to threaten humans.Vaccination is the most effective way to fight against the virus.However,due to rapid mutation of the virus,antigenic drift happens frequently,which leads to inefficiency of influenza vaccines.How to timely identify antigenic variants is an important question in influenza surveillance.

Therapeutics of Integrative Medicine Ameliorate Immunological Disorders of the Nervous System: A Meta-Analysis

Objective: The objective of the study is to evaluate the therapeutic efficacy of integrative medicine in the treatment of immunological disorders of the nervous system. Methods: The Preferred Reporting Items for Systematic Reviews and Meta-Analysis guidelines were adopted to conduct this study, which included randomized controlled trials with a confirmed diagnosis of multiple sclerosis(MS), myasthenia gravis(MG), and Guillain–Barre syndrome(GBS), all of which were treated with integrative medicine. The effective rate, recurrent frequency, and disease score were used as the markers of outcome variables for the meta-analysis. Results: A total of 48 randomized control trials were included. The effective rates of treatment with integrative medicine were noticeably higher than those with Western medicine alone for the three diseases. The recurrence frequency for MS and the recurrence rate for MG treated with integrative medicine were reduced more than those with Western medicine alone. The Extended Disability Status Scale, acetylcholine receptor antibody, and Hughes scores for MS, MG, and GBS, respectively, treated with integrative medicine were significantly lower than those with Western medicine alone. The risks of bias in the literature evaluation showed that the quality of the included studies was not high. Conclusions: Compared to treatment with Western medicine alone, integrative medicine might ameliorate immunological disorders of the nervous system.

B7-H3 confers stemness characteristics to gastric cancer cells by promoting glutathione metabolism through AKT/pAKT/Nrf2 pathway

Background:Cancer stem-like cells(CSCs)are a small subset of cells in tumors that exhibit self-renewal and differentiation properties.CSCs play a vital role in tumor formation,progression,relapse,and therapeutic resistance.B7-H3,an immunoregulatory protein,has many protumor functions.However,little is known about the mechanism underlying the role of B7-H3 in regulating gastric cancer(GC)stemness.Our study aimed to explore the impacts of B7-H3 on GC stemness and its underlying mechanism.Methods:GC stemness influenced by B7-H3 was detected both in vitro and in vivo.The expression of stemness-related markers was examined by reverse transcription quantitative polymerase chain reaction,Western blotting,and flow cytometry.Sphere formation assay was used to detect the sphere-forming ability.The underlying regulatory mechanism of B7-H3 on the stemness of GC was investigated by mass spectrometry and subsequent validation experiments.The signaling pathway(Protein kinase B[Akt]/Nuclear factor erythroid 2-related factor 2[Nrf2]pathway)of B7-H3 on the regulation of glutathione(GSH)metabolism was examined by Western blotting assay.Multi-color immunohistochemistry(mIHC)was used to detect the expression of B7-H3,cluster of differentiation 44(CD44),and Nrf2 on human GC tissues.Student’s t-test was used to compare the difference between two groups.Pearson correlation analysis was used to analyze the relationship between two molecules.The Kaplan-Meier method was used for survival analysis.Results:B7-H3 knockdown suppressed the stemness of GC cells both in vitro and in vivo.Mass spectrometric analysis showed the downregulation of GSH metabolism in short hairpin B7-H3 GC cells,which was further confirmed by the experimental results.Meanwhile,stemness characteristics in B7-H3 overexpressing cells were suppressed after the inhibition of GSH metabolism.Furthermore,Western blotting suggested that B7-H3-induced activation of GSH metabolism occurred through the AKT/Nrf2 pathway,and inhibition of AKT signaling pathway could suppress not only GSH metabolism but also GC stemness.mIHC showed that B7-H3 was highly expressed in GC tissues and was positively correlated with the expression of CD44 and Nrf2.Importantly,GC patients with high expression of B7-H3,CD44,and Nrf2 had worse prognosis(P=0.02).Conclusions:B7-H3 has a regulatory effect on GC stemness and the regulatory effect is achieved through the AKT/Nrf2/GSH pathway.Inhibiting B7-H3 expression may be a new therapeutic strategy against GC.

CCL5 deficiency promotes liver repair by improving inflammation resolution and liver regeneration through M2 macrophage polarization

Despite the diverse etiologies of drug-induced liver injury(DILI),innate immunity activation is a common feature involved in DILI progression.However,the involvement of innate immunity regulation in inflammation resolution and liver regeneration in DILI remains obscure.Herein,we identified the chemokine CCL5 as a central mediator of innate immunity regulation in the pathogenesis of DILI.First,we showed that serum and hepatic CCL5 levels are elevated in both DILI patients and an APAP-induced liver injury(AILI)mouse model.Interestingly,both nonparenchymal cells and stressed hepatocytes are cell sources of CCL5 induction in response to liver injury.Functional experiments showed that CCL5 deficiency has no effect on the early phase of AILI but promotes liver repair in the late phase mainly by promoting inflammation resolution and liver regeneration,which are associated with an increased number of hepatic M2 macrophages.Mechanistically,CCL5 can directly activate M1 polarization and impede M2 polarization through the CCR1-and CCR5-mediated activation of the MAPK and NF-κB pathways.We then showed that CCL5 inhibition mediated by either a CCL5-neutralizing antibody or the antagonist Met-CCL5 can greatly alleviate liver injury and improve survival in an AILI mouse model.Our data demonstrate CCL5 induction during DILI,identify CCL5 as a novel innate immunity regulator in macrophage polarization,and suggest that CCL5 blockage is a promising therapeutic strategy for the treatment of DILI.

Neutrophil Extracellular Traps in Autoimmune Diseases

INTRODUCTIONIn 2004, NETosis was first reported as an important step to kill bacteria by neutrophils. During the process ofN ETosis, neutrophil extracellular traps （NETs） that contain large web-like structures of decondensed chromatin decorated with histones and intracellular components, including neutrophil elastase （NE）, myeloperoxidase （MPO）, high mobility group protein B I （HMGBI）, and proteinase 3 （PR3）, are extruded into the extracellular space, The structures of NETs enable the neutrophil to potently catch and kill pathogens at the site of inflammation. Furthermore, increasing studies have identified the presence of NETs in autoimmune diseases. NETs deliver multiple autoantigens to host immtme system that induce autoimmune responses and directly release damage-associated molecular patterns to amplify inflammatory responses. Therefore, NETs are commonly described to play a crucial role in the pathogenesis and development of autoimmune diseases in recent years.