The Drug-carrier Interactions, Release Behaviors and Cell Responses of Hydroxyapaptite Containing Several Chinese Medicines

The present work shows drug-carrier interactions, release behaviors and cell responses of hydroxyapatite （HA） containing salvianolic acid B （Sal B）, astragalus polysaecharide （APS）, and naringin. X-ray diffraction （XRD） showed that the crystallinity and crystal size of HA decreased significantly when Sal B was added （p〈0.05）. Transmission electron microscope （TEM） confirmed that the nano-acicular crystals of HA containing Sal B were the most fine among all specimens. It was conjectured that Sal B preferentially adsorbed on the positively charged surface of HA crystals to inhibit their growth. In vitro release of HA containing Chinese medicines followed the first-order equation. The drug-carrier affinity between HA and Sal B might have prolonged the release of Sal B. The proliferation and differentiation of osteoblasts were promoted by Chinese medicines containing HA in the time and dosage dependent manner. The osteoblasts displayed a polygonal morphology with cell-cell junctions in all cases. It is suggested that the contained Chinese medicines would promote the activities of the osteoblasts.

FROM MOLECULAR RESPONSE TO CELL RESPONSE-Approaching the complexity of biological systems

I. THE COMPLEXITY OFBIOLOGICAL RESPONSESFor an organism, to be living or notdepends on its response to foreign matters.Facing the increasing amount and diversi-ty of chemicals, natural and synthetic, tounderstand the principles of the biologicalresponses becomes extremely importantin pursuing the way of rational utiliza-tion and governing the foreign matters.However, most biological responses aretoo complex to explore their nature. Forinstance, the risk to human beings andorganisms related to the application ofrare earths in agriculture, forestation, fish-ery and husbandry has been argued

Durable natural killer cell response after three doses of SARS-CoV-2 inactivated vaccine in HIV-infected individuals

Background:Severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)vaccine can induce a potent cellular and humoral immune response to protect against SARS-CoV-2 infection.However,it was unknown whether SARS-CoV-2 vaccination can induce effective natural killer(NK)cell response in people living with human immunodeficiency virus(PLWH)and healthy individuals.Methods:Forty-seven PLWH and thirty healthy controls(HCs)inoculated with SARS-CoV-2 inactivated vaccine were enrolled from Beijing Youan Hospital in this study.The effect of SARS-CoV-2 vaccine on NK cell frequency,phenotype,and function in PLWH and HCs was evaluated by flow cytometry,and the response of NK cells to SARS-CoV-2 Omicron Spike(SARS-2-OS)protein stimulation was also evaluated.Results:SARS-CoV-2 vaccine inoculation elicited activation and degranulation of NK cells in PLWH,which peaked at 2 weeks and then decreased to a minimum at 12 weeks after the third dose of vaccine.However,in vitro stimulation of the corresponding peripheral blood monocular cells from PLWH with SARS-2-OS protein did not upregulate the expression of the aforementioned markers.Additionally,the frequencies of NK cells expressing the activation markers CD25 and CD69 in PLWH were significantly lower than those in HCs at 0,4 and 12 weeks,but the percentage of CD16^(+)NK cells in PLWH was significantly higher than that in HCs at 2,4 and 12 weeks after the third dose of vaccine.Interestingly,the frequency of CD16^(+)NK cells was significantly negatively correlated with the proportion of CD107a^(+)NK cells in PLWH at each time point after the third dose.Similarly,this phenomenon was also observed in HCs at 0,2,and 4 weeks after the third dose.Finally,regardless of whether NK cells were stimulated with SARS-2-OS or not,we did not observe any differences in the expression of NK cell degranulation markers between PLWH and HCs.Conclusions:SARS-CoV-2 vaccine elicited activation and degranulation of NK cells,indicating that the inoculation of SARS-CoV-2 vaccine enhances NK cell immune response.

Neutralization against SARS-CoV-2 Delta/Omicron variants and B cell response after inactivated vaccination among COVID-19 convalescents

Emerging SARS-CoV-2 variants have made COVID-19 convalescents susceptible to re-infection and have raised concern about the efficacy of inactivated vaccination in neutralization against emerging variants and antigen-specific B cell response.To this end,a study on a long-term cohort of 208 participants who have recovered from COVID-19 was conducted,and the participants were followed up at 3.3(Visit 1),9.2(Visit 2),and 18.5(Visit 3)months after SARS-CoV-2 infection.They were classified into three groups(no-vaccination(n=54),one-dose(n=62),and two-dose(n=92)groups)on the basis of the administration of inactivated vaccination.The neutralizing antibody(NAb)titers against the wild-type virus continued to decrease in the no-vaccination group,but they rose significantly in the one-dose and two-dose groups,with the highest NAb titers being observed in the two-dose group at Visit 3.The NAb titers against the Delta variant for the no-vaccination,one-dose,and two-dose groups decreased by 3.3,1.9,and 2.3 folds relative to the wild-type virus,respectively,and those against the Omicron variant decreased by 7.0,4.0,and 3.8 folds,respectively.Similarly,the responses of SARS-CoV-2 RBD-specific B cells and memory B cells were boosted by the second vaccine dose.Results showed that the convalescents benefited from the administration of the inactivated vaccine(one or two doses),which enhanced neutralization against highly mutated SARS-CoV-2 variants and memory B cell responses.Two doses of inactivated vaccine among COVID-19 convalescents are therefore recommended for the prevention of the COVID-19 pandemic,and vaccination guidelines and policies need to be updated.

SARS-CoV-2-specific T cell responses wane profoundly in convalescent individuals 10 months after primary infection

A key question in the coronavirus disease 2019(COVID-19)pandemic is the duration of specific T cell responses against the severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)post primary infection,which is difficult to address due to the large-scale COVID-19 vaccination and re-exposure to the virus.Here,we conducted an analysis of the long-term SARS-CoV-2-specific T cell responses in a unique cohort of convalescent individuals(CIs)that were among the first to be infected worldwide and without any possible antigen re-exposure since then.The magnitude and breadth of SARS-CoV-2-specific T cell responses correlated inversely with the time that had elapsed from disease onset and the age of those CIs.The mean magnitude of SARS-CoV-2-specific CD4 and CD8 T cell responses decreased about 82%and 76%,respectively,over the time period of ten months after infection.Accordingly,the longitudinal analysis also demonstrated that SARS-CoV-2-specific T cell responses waned significantly in 75%of CIs during the follow-up.Collectively,we provide a comprehensive characterization of the long-term memory T cell response in CIs,suggesting that robust SARS-CoV-2-specific T cell immunity post primary infection may be less durable than previously expected.

Human Leukocyte Antigen-A Allele Distribution in Nasopharyngeal Carcinoma Patients Showing Anti-Melanoma-Associated Antigen A or Synovial Sarcoma X-2 T Cell Response in Blood

Background： Development of innovative immunotherapy is imperative to improve the poor survival of the nasopharyngeal carcinoma （NPC） patients. In this study, we evaluated the T cell response to melanoma-associated antigen （MAGE）-A1, MAGE-A3, or synovial sarcoma X-2 （SSX-2） in the peripheral blood of treatment-naive NPC patients. The relationship of responses among the three proteins and the human leukocyte antigen （HLA）-A types were analyzed to provide evidence of designing novel therapy. Methods： Sixty-one NPC patients admitted into the Tumor Hospital affiliated to the Xinjiang Medical University between March 2015 and July 2016 were enrolled. Mononuclear cells were isolated from the peripheral blood before any treatment. HLA-A alleles were typed with Sanger sequence-based typing technique. The T cell response to the MAGE-A1, MAGE-A3, or SSX-2 was evaluated with the Enzyme-Linked ImmunoSpot assay. Mann-Whitney U-test was used to compare the T cell responses from different groups. Spearman＇s rank correlation was used to analyze the relationship of T cell responses. Results： HLA-A＊02：01, A＊02：07, and A＊24：02 were the three most frequent alleles （18.9%, 12.3%, and 11.5%, respectively） among the 22 detected alleles. 31.1%, 19.7%, and 16.4% of the patients displayed MAGE-A1, MAGE-A3, or SSX-2-specific T cell response, respectively. The magnitudes of response to the three proteins were 32.5, 38.0, and 28.7 SFC/106 peripheral blood mononuclear cells, respectively. The T cell response against the three proteins correlated with each other to different extent. The percentage of A＊02：01 and A＊24：02 carriers were significantly higher in patients responding to any of the three proteins compared to the nonresponders. Conclusion： MAGE-A1, MAGE-A3, or SSX-2-specific T cell responses were detectable in a subgroup of NPC patients, the frequency and magnitude of which were correlated.

An adjusted ELISpot-based immunoassay for evaluation of SARS-CoV-2-specific T-cell responses

Like antibody evaluation,using an effective antigen‐specific T‐cell immunity assessment method in coronavirus disease 2019(COVID‐19)patients,survivors and vaccinees is crucial for understanding the immune persistence,prognosis assessment,and vaccine development for COVID‐19.This study evaluated an empirically adjusted enzyme‐linked immunospot assay for detecting severe acute respiratory syndrome coronavirus 2(SARS‐CoV‐2)‐specific T‐cell immunity in 175 peripheral blood samples from COVID‐19 convalescents and healthy individuals.Results of viral nucleic acid were used as the gold standard of infection confirmation.The SARS‐CoV‐2M peptide pool had higher sensitivity of 85%and specificity of 71%for the single peptide pool.For combined peptide pools,the parallel evaluation(at least one of the peptide pools is positive)of total peptide pools(S1&S2&M&N)had higher sensitivity(up to 93%),and the serial evaluation(all peptide pools are positive)of total peptide pools had higher specificity(up to 100%).The result of the serial evaluation was better than that of the parallel evaluation as a whole.The detection efficiency of M and N peptide pool serial evaluation appeared the highest,with a sensitivity of 80%and specificity of 93%.This T‐cell immunity detection assay introduced in this report can achieve high operability and applicability.Therefore,it can be an effective SARS‐CoV‐2‐specific cellular immune function evaluation method.

Enhance Stability and in vitro Cell Response to a Bioinspired Coating on Zr Alloy with Increasing Chitosan Content

The aim of the present paper is to characterize bioinspired chitosan （CS） ＋ hydroxyapatite （HA） coatings with various components ratio on a zirconium alloy with titanium. The coatings were characterized by FT-IR, SEM, hydrophilic/hydrophobic balance, adherence, roughness, electrochemical stability and in vitro cell response. Electrochemical tests, including potentio- dynamic polarization curves and electrochemical impedance spectroscopy, were performed in normal saline physiological solution. Cell viability of MC3T3-E1 osteoblasts, lactate dehydrogenase, nitric oxide, and Reactive Oxygen Species （ROS） levels, as well as actin cytoskeleton morphology, were evaluated as biological in vitro tests. The results on in vitro cell response indicated good cell membrane integrity and viability for all samples, but an increased cell number, a decreased ROS level and a better cytoskeleton organization were noticed for the sample with a higher CS content. The coating with highest CS concen- tration indicated the best performance based on the experimental data. The highest hydrophilic character, highest resistance to corrosion and best biocompatibility as well recommend this coating for bioapplications in tissue engineering.

Mild Cytokine Elevation, Moderate CD4^+ T Cell Response and Abundant Antibody Production in Children with COVID-19

Children with Coronavirus Disease 2019(COVID-19) were reported to show milder symptoms and better prognosis than their adult counterparts, but the difference of immune response against SARS-CoV-2 between children and adults hasn’t been reported. Therefore we initiated this study to figure out the features of immune response in children with COVID-19.Sera and whole blood cells from 19 children with COVID-19 during different phases after disease onset were collected.The cytokine concentrations, SARS-CoV-2 S-RBD or N-specific antibodies and T cell immune responses were detected respectively. In children with COVID-19, only 3 of 12 cytokines were increased in acute sera, including interferon(IFN)-cinduced protein 10(IP10), interleukin(IL)-10 and IL-16. We observed an increase in T helper(Th)-2 cells and a suppression in regulatory T cells(Treg) in patients during acute phase, but no significant response was found in the IFN-cproducing or tumor necrosis factor(TNF)-a-producing CD8?T cells in patients. S-RBD and N IgM showed an early induction, while S-RBD and N IgG were prominently induced later in convalescent phase. Potent S-RBD IgA response was observed but N IgA seemed to be inconspicuous. Children with COVID-19 displayed an immunophenotype that is less inflammatory than adults, including unremarkable cytokine elevation, moderate CD4?T cell response and inactive CD8?T cell response, but their humoral immunity against SARS-CoV-2 were as strong as adults. Our finding presented immunological characteristics of children with COVID-19 and might give some clues as to why children develop less severe disease than adults.

The HMGB1 signaling pathway activates the inflammatory response in Schwann cells

Schwann cells are not only myelinating cells, but also function as immune cells and express numerous innate pattern recognition receptors, including the Toll-like receptors. Injury to peripheral nerves activates an inflammatory response in Schwann cells. However, it is unclear whether specific endogenous damage-associated molecular pattern molecules are involved in the inflammatory response following nerve injury. In the present study, we demonstrate that a key damage-associated molecular pattern molecule, high mobility group box 1（HMGB1）, is upregulated following rat sciatic nerve axotomy, and we show colocalization of the protein with Schwann cells. HMGB1 alone could not enhance expression of Toll-like receptors or the receptor for advanced glycation end products（RAGE）, but was able to facilitate migration of Schwann cells. When Schwann cells were treated with HMGB1 together with lipopolysaccharide, the expression levels of Toll-like receptors and RAGE, as well as inflammatory cytokines were upregulated. Our novel findings demonstrate that the HMGB1 pathway activates the inflammatory response in Schwann cells following peripheral nerve injury.

Subversion of cellular stress responses by poxviruses

Cellular stress responses are powerful mechanisms that prevent and cope with the accumulation of macromolecular damage in the cells and also boost host defenses against pathogens. Cells can initiate either protective or destructive stress responses depending, to a large extent, on the nature and duration of the stressing stimulus as well as the cell type. The productive replication of a virus within a given cell places inordinate stress on the metabolism machinery of the host and, to assure the continuity of its replication, many viruses have developed ways to modulate the cell stress responses. Poxviruses are among the viruses that have evolved a large number of strategies to manipulate host stress responses in order to control cell fate and enhance their replicative success. Remarkably, nearly every step of the stress responses that is mounted during infection can be targeted by virally encoded functions. The fine-tuned interactions between poxviruses and the host stress responses has aided virologists to understand specific aspects of viral replication; has helped cell biologists to evaluate the role of stress signaling in the uninfected cell; and has tipped immunologists on how these signals contribute to alert the cells against pathogen invasionand boost subsequent immune responses. This review discusses the diverse strategies that poxviruses use to subvert host cell stress responses.

Measurement for Spectral Response of Solar Cells

General Research Institute for Non-ferrous Metals in cooperation with Changchun Instituteof Optical and Fine Mechanics prepared an automatic measurement instrument for spectral re-sponse test of solar cells. This instrument having enough accuracy can measure at AM1.5 condi-tion. It meets the needs for measurement of spectral response for silicon, gallium arsenide solarcells.

Cytotoxic Responses and Apoptosis in Rat Kidney Epithelial Cells Exposed to Lead

The toxic effects of lead on normal rat kidney epithelial cells（NRK cells）may occur via various pathways.However,the role of intrinsic mitochondrial pathway in Lead-induced apoptosis in NRK cells has not been investigated.The purpose of our study was to investigate cytotoxic responses and cell apoptosis mediated by lead in NRK cells.

Expression and significance of toll-like receptor 2,4 in peripheral blood mononuclear cells in patients with systemic inflammatory response syndrome

To explore changes of toll-like receptor (TLR) 2,4 in peripheral blood mononuclear cells (PBMC) in acute abdomen patients with systemic inflammatory response syndrome (SIRS) and their significance.Methods A clinical study was done on 103 patients of which 65 were with SIRS.The mRNA expression of TLR2,4 were detected by RT-PCR;the expression of TNF-α and IL-6 were observed by ELISA;the correlation between TLR2,4 mRNA,the level of TNF-α and IL-6,and the clinical course was evaluated.Results TLR2 mRNA ,TNF-α and IL-6 were upregulated markedly on the first day of hospitalization,then decreased gradually;TLR2 mRNA maintained on high level till the 5th day.The expression of TLR2,4 mRNA was positive correlated with the level of TNF-α and IL-6,and the length of stay.TLR2,4 mRNA expression increased in patients with multiple organ failure.Conclusion In actue abdomen patients with SIRS,the expression of TLR2,4 of PBMC increased markedly,indicating its improtant role in the pathogenesis of SIRS.4 refs,2 figs,2 tabs.

Natural course of chronic hepatitis B is characterized by changing patterns of programmed death type-1 of CD8-positive T cells

AIM:To investigate if and how programmed death type-1(PD-1)expression affects the natural course of hepatitis B virus(HBV)infection. METHODS:Sixty-four patients in different natural stages of chronic HBV infection were enrolled in this study.PD-1 expression in total T cells was detected by flow cytometry.Levels of total CD8+T cell responses and proliferation in relation to PD-1 expression levels were analyzed with intracellular staining and PD-1/ PD-L1 blockage. RESULTS:The PD-1 expression in T cells was dynamically changed during the natural course of chronic HBV infection,did not significantly increase in the immune tolerance phase,and returned to normal in the inactive virus carrier stage.Blockage of the PD-1/PD-L1 pathway could not affect the T-cell response in the immune tolerance and inactive virus carrier stages of chronic HBV infection.However,it could significantly restore the T-cell response in the immune clearance stage of chronic HBV infection.Furthermore,the PD-1 expression level in T cells was associated with the alanine aminotransferase level during the immune clearance stage of chronic HBV infection. CONCLUSION:The PD-l/PD-L1 pathway plays a different role in T-cell response during the natural course of chronic HBV infection.

Immune Responses in Wild-type Mice Against Prion Proteins Induced Using a DNA Prime-Protein Boost Strategy

Objective To break immune tolerance to prion （PrP） proteins using DNA vaccines.Methods Four different human prion DNA vaccine candidates were constructed based on the pcDNA3.1 vector：PrP‐WT expressing wild‐type PrP,Ubiq‐PrP expressing PrP fused to ubiquitin,PrP‐LII expressing PrP fused to the lysosomal integral membrane protein type II lysosome‐targeting signal,and PrP‐ER expressing PrP locating the ER.Using a prime‐boost strategy,three‐doses of DNA vaccine were injected intramuscularly into Balb/c mice,followed by two doses of PrP protein.Two weeks after the last immunization,sera and spleens were collected and PrP‐specific humoral and cellular immune responses evaluated by ELISA and ELISPOT tests.Results Higher levels of serum PrP antibodies were detected in mice vaccinated using the strategy of DNA priming followed by protein boosting.Of these,WT‐PrP,Ubiq‐PrP,and PrP‐LII induced significantly higher humoral responses.ELISPOT tests showed markedly increased numbers of IFN‐γ‐secreting T cells in mice vaccinated using the strategy of DNA priming followed by protein boosting after stimulation with recombinant PrP23‐90 and PrP23‐231.PrP‐ER inducedthe strongest T‐cell response.Conclusion Prion vaccines can break tolerance to PrP proteins and induce PrP‐specific humoral and cellular immune responses.

Preventive effects of Bifidobacterium lactis Probio-M8 on ovalbumin-induced food allergy in mice

Food allergy is a significant public health concern globally.Certain probiotics have been found to enhance food allergy by regulating immune-microbe interactions in animal models and patients.However,the effects of Bifidobacterium lactis Probio-M8 on food allergy have not been thoroughly investigated.The present study examined the anti-allergic properties of Probio-M8,particularly in relation to immune response and gut microbiota composition.Results demonstrate that oral administration of Probio-M8 effectively mitigated the allergy symptoms triggered by ovalbumin(OVA)by ameliorating the morphological damage in the jejunum,reducing OVA-specific IgE and histamine levels in the serum,and suppressing Th2 cytokines(interleukin(IL)4 and IL-13)while increasing Th1 cytokines(interferon(IFN)γ)and regulatory T(Treg)cytokines(IL-10 and transforming growth factor(TGF)β1)in the culture supernatants of splenic cells.Furthermore,Probio-M8 effectively altered the diversity and composition of gut microbiota,particularly the relative abundances of Akkermansia_muciniphila in OVA-induced mice.Compared to the OVA group,the Probio-M8 group showed a decrease in the relative abundance of Akkermansia_muciniphila.In conclusion,Probio-M8 demonstrates the potential to alleviate food allergy by regulating the Th1/Th2 response and modulating gut microbiota,thereby offering a novel therapeutic strategy for patients with food allergy.

HIV-Specific IL-2^+ and/or IFN-γ^+ CD8^+ T Cell Reponses during Chronic HIV-1 Infection in Former Blood Donors

Objective Conflicting data have been generated from previous studies to determine which kind of relationship exists between HIV-1 specific CD8 Tcell responses and HIV-1 viral load or CD4 count over the course of infection.In this study,153 HIV-1 infected LTNPs were enrolled to investigate the role of HIV-1 specific CD8 T-cell responses in chronic HIV-1 infection among HIV-1 infected former blood donors.Methods The patients were stratified into three groups according to CD4 count：CD4≥500 cells/μL;350 cells/μL≤CD4〈500 cells/μL;CD4〈350 cells/μL.PBMCs were isolated from the patients＇ anticoagulated blood samples.IL-2 and IFN-γ secretions of CD 8 T cells against 17 HIV-1 consensus B full peptide pools were analyzed by using ICS assay.Results An overall inverse correlation were observed between CD4 count and plasma viral load.Although no significant difference was observed during the comparisons of frequency/breadth of HIV-1 specific CD8 T cell responses,CD4 count stratification analysis showed that different correlation pattern existed in three strata：as for patients whose CD4 counts were less than 350 cells/μL,no significant correlations were identified between frequency/breadth of HIV-1 specific CD8 T cell responses and CD4 count/viral load;as for patients whose CD4 counts ranged from 350 cells /μL to 500 cells/μL,significant correlation was only observed between the response breadth of IL-2＋IFN-γ＋ CD8 T cells and CD4 count;however,as for patients whose CD4 counts were more than 500 cells/μL,direct correlations were identified between IL-2＋IFN-γ＋/IL-2＋/IFN-γ＋ CD8 T cells and viral load or CD4 count.Conclusions Universal consistent inverse correlation was only indentified between CD4 count and viral load.The relationship between HIV-1 specific CD8 T cell responses and CD4 count/viral load varied in different CD4 strata,which showed that better preserved CD4 T cells were correlated with better CD8 T cell functions.

Induced Th2 dominant immune response in APPswe, PSEN1dE9 transgenic mice after nasal immunization with an adenoviral vector encoding 10 tandem repeats of beta-amyloid 3-10

Immunotherapy for Alzheimer＇s disease （AD） is effective in improving cognitive function in transgenic mouse models of AD. Because the AN1792 [beta-amyloid （Aβ） 1-42] vaccine was halted because of T cell mediated meningoencephalitis, many scientists are searching for a nove） vaccine to avoid the T cell mediated immune response caused by the Aβ1-42. Importantly, the time when the immunization is begun can influence the immune effect. In this study, an adenovirus vaccine was constructed containing 10 x Aβ3-10 repeats and gene adjuvant CpG DNA. Transgenic AD mice were immunized intranasally for 3 months. After 10 × Aβ3-10 vaccine immunization, high titers of anti-Aβ42 IgG1 predominant antibodies were induced. In spatial learning ability and probe tests, the 10 × Aβ3-10 immunized mice showed significantly improved memories compared to control mice. The 10 × Aβ3-10 vaccine resulted in a robust Th2 dominant humoral immune response and reduced learning deficits in AD mice. In addition, the 10 × Aβ3-10 vaccine might be more efficient if administered before Aβ aggregation at an early stage in the AD mouse brain. Thus, the adenovirus vector encoding 10 × Aβ-10 is a promising vaccine for AD.