Altered HLA-A2-restricted TP53 epitope induces specific CTL cytotoxicity against hepatocellular carcinoma.

High-frequency mutation of the TP53 tumor suppressor gene is observed in multiple human cancers, which promotes cancer progression. However, the mutated gene-encoded protein may serve as a tumor antigen to elicit tumor-specific immune responses. In this study, we detected widespread expression of shared TP53-Y220C neoantigen in hepatocellular carcinoma with low affinity and low stability of binding to HLA-A0201 molecules. We substituted the amino acid sequences VVPCEPPEV with VLPCEPPEV in the TP53-Y220C neoantigen to yield a TP53-Y220C (L2) neoantigen. This altered neoantigen was found to increase affinity and stability and induce more cytotoxic T lymphocytes (CTLs), indicating improvements in immunogenicity. In vitro assays showed the cytotoxicity of CTLs stimulated by both TP53-Y220C and TP53-Y220C (L2) neoantigens against multiple HLA-A0201-positive cancer cells expressing TP53-Y220C neoantigens; however, the TP53-Y220C (L2) neoantigen showed higher cytotoxicity than the TP53-Y220C neoantigen against cancer cells. More importantly, in vivo assays demonstrated greater inhibition of hepatocellular carcinoma cell proliferation by TP53-Y220C (L2) neoantigen-specific CTLs relative to TP53-Y220C neoantigen in zebrafish and nonobese diabetic/severe combined immune deficiency mouse models. The results of this study demonstrate enhanced immunogenicity of the shared TP53-Y220C (L2) neoantigen, which has the potential as dendritic cells or peptide vaccines for multiple cancers.

An altered HLA-A0201-restricted MUC1 epitope that could induce more efficient anti-tumor effects against gastric cancer.

MUC1 is a tumor-associated antigen (TAA) overexpressed in many tumor types, which makes it an attractive target for cancer immunotherapy. However, this marker is a non-mutated antigen without high immunogenicity. In this study, we designed several new altered peptides by replacing amino acids in their sequences, which were derived from a low-affinity MUC1 peptide, thus bypassing immune tolerance. Compared to the wild-type (WT) peptide, the altered MUC1 peptides (MUC11081-1089L2, MUC11156-1164L2, MUC11068-1076Y1) showed higher affinity to the HLA-A0201 molecule and stronger immunogenicity. Furthermore, these altered peptides resulted in the generation of more cytotoxic T lymphocytes (CTLs) that could cross-recognize gastric cancer cells expressing WT MUC1 peptides, in an HLA-A0201-restricted manner. In addition, M1.1 (MUC1950-958), a promising antitumor peptide that has been tested in multiple tumors, was not able to induce stronger antitumor responses. Collectively, our results demonstrated that altered peptides from MUC1, as potential HLA-A0201-restricted CTL epitopes, could serve as peptide vaccines or constitute components of peptide-loaded dendritic cell vaccines for gastric cancer treatment.

Secondary Immunodeficiency and Hypogammaglobulinemia with IgG Levels of <5 g/L in Patients with Multiple Myeloma: A Retrospective Study Between 2012 and 2020 at a University Hospital in China.

BACKGROUND Infections are the main cause of mortality and morbidity in multiple myeloma (MM) patients. However, adult immunodeficiency specialists in China are lacking, and the care of secondary immunodeficiency (SID) and the prognostic role of hypogammaglobulinemia in MM is unknown. MATERIAL AND METHODS MM patients (295) were retrospectively analyzed between January 2012 and 2020 in Zhejiang Provincial People's Hospital, Hangzhou Medical College. MM patients with immunoglobulin (Ig) G <5 g/L were defined as SID patients. The care of these patients and the prognostic role of IgG <5 g/L were analyzed RESULTS Forty-five of 295 MM patients with IgG <5 g/L were defined as SID patients. These 45 patients mainly had recurrent infections, especially pulmonary bacterial infections; 2 patients had them 5 times/year. The median survival time was significantly shorter in MM patients with SID (24 vs 66 months). More importantly, the multivariate and univariate analysis revealed that IgG <5 g/L was an independent prognostic factor for MM patients. CONCLUSIONS Ig replacement therapy or prophylactic antibiotics for MM patients with SID were lacking in this single retrospective study. IgG <5 g/L could be a prognostic marker for MM patients.

Mitochondrial ROS Release and Subsequent Akt Activation Potentially Mediated the Anti-Apoptotic Effect of a 50-Hz Magnetic Field on FL Cells.

BACKGROUND/AIMS: Our previous study showed that exposure to a 50-Hz magnetic field (MF) could induce transient mitochondrial permeability transition (MPT) in cells. In the present study, the aim was to explore the possible biological implications of MF-induced transient MPT. MATERIALS AND METHODS: Human amniotic (FL) cells were exposed to MF for different durations or intensities followed by incubation with staurosporine for 4 h. After MF exposure, cell early apoptosis, cell viability mitochondrial ROS and the level of phosphorylated Akt were assessed. After MF exposure followed by incubation with staurosporine, cell early apoptosis was assessed. RESULTS: MF exposure had a protective effect against early apoptosis induced by staurosporine, which could be abolished by MPT inhibitors, although MF exposure alone had no significant effect on early apoptosis or viability of cells. In addition, exposing cells to MF increased the level of mitochondrial ROS which were released into cytoplasm through mitochondrial permeability transition pores (mPTP), and induced ROS-dependent phosphorylation of Akt. Furthermore, the anti-apoptotic effect of MF exposure was completely eliminated when Akt was inhibited. CONCLUSIONS: The present study indicated a possibility that mitochondrial ROS release through mPTP and subsequent Akt activation were necessary for the anti-apoptotic effect of MF.

Specific mortality in patients with diffuse large B-cell lymphoma: a retrospective analysis based on the surveillance, epidemiology, and end results database.

BACKGROUND: The full potential of competing risk modeling approaches in the context of diffuse large B-cell lymphoma (DLBCL) patients has yet to be fully harnessed. This study aims to address this gap by developing a sophisticated competing risk model specifically designed to predict specific mortality in DLBCL patients. METHODS: We extracted DLBCL patients' data from the SEER (Surveillance, Epidemiology, and End Results) database. To identify relevant variables, we conducted a two-step screening process using univariate and multivariate Fine and Gray regression analyses. Subsequently, a nomogram was constructed based on the results. The model's consistency index (C-index) was calculated to assess its performance. Additionally, calibration curves and receiver operator characteristic (ROC) curves were generated to validate the model's effectiveness. RESULTS: This study enrolled a total of 24,402 patients. The feature selection analysis identified 13 variables that were statistically significant and therefore included in the model. The model validation results demonstrated that the area under the receiver operating characteristic (ROC) curve (AUC) for predicting 6-month, 1-year, and 3-year DLBCL-specific mortality was 0.748, 0.718, and 0.698, respectively, in the training cohort. In the validation cohort, the AUC values were 0.747, 0.721, and 0.697. The calibration curves indicated good consistency between the training and validation cohorts. CONCLUSION: The most significant predictor of DLBCL-specific mortality is the age of the patient, followed by the Ann Arbor stage and the administration of chemotherapy. This predictive model has the potential to facilitate the identification of high-risk DLBCL patients by clinicians, ultimately leading to improved prognosis.

Co-exposure of decabromodiphenyl ethane and cadmium increases toxicity to earthworms: Enrichment, oxidative stress, damage and molecular binding mechanisms.

The increase of electronic waste worldwide has resulted in the exacerbation of combined decabromodiphenyl ethane (DBDPE) and cadmium (Cd) pollution in soil, posing a serious threat to the safety of soil organisms. However, whether combined exposure increases toxicity remains unclear. Therefore, this study primarily investigated the toxic effects of DBDPE and Cd on earthworms at the individual, tissue, and cellular levels under single and combined exposure. The results showed that the combined exposure significantly increased the enrichment of Cd in earthworms by 50.32-90.42 %. The toxicity to earthworms increased with co-exposure, primarily resulting in enhanced oxidative stress, inhibition of growth and reproduction, intensified intestinal and epidermal damage, and amplified coelomocyte apoptosis. PLS-PM analysis revealed a significant and direct relationship between the accumulation of target pollutants in earthworms and oxidative stress, damage, as well as growth and reproduction of earthworms. Furthermore, IBR analysis indicated that SOD and POD were sensitive biomarkers in earthworms. Molecular docking elucidated that DBDPE and Cd induced oxidative stress responses in earthworms through the alteration of the conformation of the two enzymes. This study enhances understanding of the mechanisms behind the toxicity of combined pollution and provides important insights for assessing e-waste contaminated soils.

Surveillance for adverse events following immunization with DTaP-containing combination vaccines in Linping, China, 2019-2022.

Background: The DTaP-Hib and DTaP-IPV/Hib combination vaccine can be used as a substitute for the diphtheria, tetanus, and acellular pertussis combined vaccine (DTaP). We aimed to evaluate the safety of multi-component vaccines containing DTaP by analyzing the reporting rates and characteristics of adverse events following immunization (AEFIs) in Linping District during the years 2019 to 2022. Methods: We obtained data of AEFI and vaccination from the National AEFI Surveillance System of China and Zhejiang Municipal Immunization Information Management System, respectively, during 2019-2022 for a descriptive, epidemiological analysis. Results: The total number of AEFI reported following vaccinations with DTaP-containing combination vaccines was 802 in Linping District from 2019 to 2022. The overall reporting rates of AEFIs following DTaP, DTaP-Hib, and DTaP-IPV/Hib vaccinations were 445.72 (537 cases), 536.29 (45 cases), and 306.13 (220 cases) per 100,000 doses in Linping District from 2019 to 2022, respectively. Only one case of a serious AEFI following DTaP vaccination, with a reporting rate of 0.83 per 100,000 doses. The composition ratio of vaccine product-related reactions for DTaP, DTaP-Hib, and DTaP-IPV/Hib were 99.81, 97.78, and 100.00%, respectively. The composition ratio of coincidental events for DTaP and DTaP-Hib were 0.19 and 2.22%, respectively. The reporting rates of total AEFIs for DTaP-IPV/Hib were lower than for DTaP. The reporting rate of local induration for DTaP-Hib was lower than for DTaP, and the reporting rates of local redness & swelling and local induration for DTaP-IPV/Hib were both lower than for DTaP. DTaP-IPV/Hib had a higher proportion of AEFIs in first quarter compared to DTaP. The reporting rate after the second dose of DTaP-Hib was higher than that of DTaP, and the reporting rates of AEFIs after the first dose and third dose of DTaP-IPV/Hib were lower than DTaP. Conclusion: The reported AEFIs to multi-component vaccines containing DTaP components during 2019-2022 in Linping District were mainly mild vaccine reactions. DTaP-containing combination vaccines demonstrated a good safety profile.

Dysregulated Tfr/Tfh2 cells in patients with polycystic ovarian syndrome.

Recent research revealed the pathogenic role of B cells in the pathogenesis of polycystic ovary syndrome (PCOS), while the Tfh cell plays a critical role in the B cell mediated autoantibody production and humoral immunity, but had not been investigated in PCOS patients. The frequency of Tfh and B cell subsets (Tfh1, Tfh2, Tfh17, naïve B, memory B, and plasma cells) in the peripheral blood of 21 PCOS patients and 15 healthy controls were investigated by flow cytometry. And the levels of follicle-stimulating hormone, luteinizing hormone, testosterone, prolactin and estradiol progesterone were measured by using the immunoluminescence method. Also, the associations between these hormone levels and Tfh cell subsets or B cell subsets were analyzed. No significant difference was observed in total Tfh cells between 21 PCOS patients and 15 healthy controls (p > 0.05). But the percentages of Tfh2 and plasma cells were significantly higher in 21 PCOS patients compared to 15 healthy controls (p < 0.05). In contrast, the frequency of Tfr cells and Tfr/Tfh2 ratio were significantly lower than healthy controls (p < 0.01). Importantly, among these cells, only the percentage of Tfh2 cells was positively correlated with the levels of testosterone (r = 0.513, p = 0.018). And the percentage of Tfr cells and Tfr/Tfh2 ratio were also positively correlated with the levels of testosterone (r = 0.567, p = 0.007; r = 0.434, p = 0.05) and prolactin (r = 0.511, p = 0.018; r = 0.490, p = 0.024). These new findings provide unique insights into dysregulated Tfh/Tfr cells in mediating the immunopathogenesis of PCOS patients.

Polylactic acid microplastics induce higher biotoxicity of decabromodiphenyl ethane on earthworms (Eisenia fetida) compared to polyethylene and polypropylene microplastics.

Decabromodiphenyl ethane (DBDPE) and microplastics (MPs), such as fossil-based polymers polyethylene (PE), polypropylene (PP), and bio-based plastics polylactic acid (PLA) are abundant in e-waste dismantling areas. However, the information on the effects of DBDPE combined with MPs (DBDPE-MPs) on earthworms is still limited. In this study, we explored the impacts of DBDPE-MPs on neurotoxic biomarkers, tissue damage, and transcriptomics of Eisenia fetida by simulating different exposure patterns of 10 mg kg-1 DBDPE and 10 mg kg-1 DBDPE-MPs (PLA, PP, and PE). Results showed that the activities of acetylcholinesterase, Na+/K+-ATPase, Ca2+/Mg2+-ATPase, carboxylate enzyme, and the contents of calcium and glutamate were significantly stimulated. DBDPE-MP co-exposure caused more severe damage to the epidermis, muscles, and tissues. Transcriptomic analysis revealed that differentially expressed genes (DEGs) of DBDPE-MPs were mainly related to inflammation, the immune system, digestive system, endocrine system, and metabolism. DBDPE and PP-MPs had similar influences on immunity and metabolism. However, DBDPE-PLA and DBDPE-PE further affected the endocrine system and signaling pathways. Specific DEGs showed that detoxification systems in the case of MPs were significantly upregulated. The study indicated that MPs exacerbated DBDPE toxicity in the nervous system, epidermis, and gene regulation of E. fetida, helping to assess the ecological risks of e-wastes and microplastics in soil.

Occurrence, spatial distribution, and risk assessment of brominated flame retardants in farmland soils of typical provinces in China.

In the present study, we investigated the occurrence, distribution, and potential risks of 4 brominated flame retardants in farmland soils across 18 provinces of China. The total mean concentrations of the BFRs were in order as DBDPE > BDE209 > HBB > TBB. DBDPE concentration was highest at 177.208 ng/kg, revealing its long-term use and persistence across the study areas. In parts of China, DBDPE was highest in the south (Sichuan, Shaanxi and Guangdong provinces), BDE209 was highest in the south (Sichuan province) and north (Jilin province), while HBB was highest in the south (Sichuan province) and east (Anhui and Zhejiang provinces) of China. Comparisons of the results in this study to other reported studies in different regions indicated that the studied BFRs concentrations were higher in the studied provinces of China. Pearson correlation between BFRs revealed both positive and negative associations within the BFRs groups and between BFRs and soil properties (SOM, CEC, pH, EC and PS%). The mean hazard quotients (HQ) of ecological risks ranged from 8.76 × 10-6 to 1.16 × 10-2 (HQ < 1) while non-carcinogenic human health risk evaluation for adults ranged from 7.05 × 10-7 - 7.48 × 10-4 (HQ < 1) and for children 2.99 × 10-4 - 4.30 × 10-2 (HQ < 1). Although the risk evaluations of BFRs from farmland soils in this study were low, the results serve as useful indicators of potential cumulative and long-term threats of BFR to rural areas where there is a high conversion of agricultural lands to non-agricultural use.

Advanced hormone receptor-positive/human epidermal growth factor receptor 2-positive invasive ductal carcinoma with cecal metastasis: A case report.

The incidence of gastrointestinal metastases from breast cancer (BC) is low. We report a special case of Luminal B (Hormone Receptor positive [HR+]/Human Epidermal Growth Factor receptor 2-positive [HER-2+]) BC. The patient presented with asymptomatic brain metastases two years after radical surgery for modified breast cancer and developed right lower abdominal pain during relief therapy. Electronic gastroenteroscopy revealed inflammatory changes in the cecal mucosa. These changes were confirmed on pathology to be cecal metastasis from BC. The patient's condition was stabilised after treatment with an antibody-drug conjugate (ADC). For patients with BC who develop appendicitis-like symptoms after treatment for invasive ductal carcinoma of the breast, clinicians should be fully aware that the possibility of cecal metastasis needs to be considered, despite the very low probability of occurrence.

Research on the policy synergy of supply-side and demand-side environmental policies: an analysis based on green value chain of manufacturing enterprises.

An enterprise's green value chain comprises a multitude of factors and links for a successful green transformation, and relevant environmental policies are multi-dimensional and multi-sectional. Environmental policies can affect the enterprise's green development through stimulating the supply of green services or by increasing the demand for green products. Based on 346 policy statements issued by various government departments from 2000 to 2020, this paper matches the "type" of environmental policy with the "dimension" of value chain of manufacturing companies, establishing a coordination model and a co-existing network to measure how both elements interact. The results indicate a notable phenomenon with the Chinese government's enterprise environmental policies, which can be described as "heavy supply-driven, light demand-pull." A strong emphasis is put on the supply side of policies, in particular for the technology innovation sector, but the demand end of policies is comparatively minimal. The synergy between the policies and the links in the green value chain is divergent, which replicates the "heavy end-treatment, light front-prevention" seen with existing policies. Policy synergy at the end of the value chain such as the green production process is more mature and well-integrated, while the synergy for green product development lacks policies on both ends. The mismatch and imbalance of policies limits the overall level of green transformation for manufacturing enterprises.

Role of Tfh17 cells in patients with multiple myeloma.

Objective Follicular helper T cells (Tfh) drive proliferation and differentiation of B cells into plasma cells, leading to antibody production; however, their role in multiple myeloma (MM) is unknown. We aimed to determine the alteration of Tfh subsets and their clinical significance in patients with MM.Method Forty-nine patients with MM were recruited in this study, including 12 newly diagnosed patients, 10 relapsed patients, and 8 patients who received autologous hematopoietic stem cell transplantation (ASCT) from Zhejiang Provincial People's Hospital. Total CD4 + CXCR5 + CD25lowCD127intermediate-high Tfh cells, CXCR3 + CCR6-Tfh1 cells, CXCR3-CCR6-Tfh2 cells, and CXCR3-CCR6 + Tfh17 cells from the peripheral blood of these patients were analyzed by flow cytometry.Result Although total Tfh cells were not significantly changed in patients with MM compared to that in healthy controls (HCs), the Tfh17/Tfh ratio was significantly elevated in patients with MM compared to that in HCs (P = 0.0001). Importantly, relapsed patients had higher Tfh17/Tfh ratio than the newly diagnosed patients (P = 0.0077). Moreover, the Tfh17/Tfh ratio was significantly decreased in patients with MM who received ASCT (post-ASCT) when compared to that in HCs and non-ASCT patients (P < 0.0001), but no change was observed between post-ASCT patients and HCs (P = 0.7498).Conclusion The Tfh17/Tfh ratio was significantly elevated in patients with MM, especially in relapsed patients, indicating that Tfh17 cells may play a critical role in the clinical progression of MM.

Altered MUC1 epitope-specific CTLs: A potential target for immunotherapy of pancreatic cancer.

The efficacy of conventional treatments for pancreatic cancer remains unsatisfactory, and immunotherapy is an emerging option for adjuvant treatment of this highly deadly disorder. The tumor-associated antigen (TAA) MUC1 is expressed in a variety of human cancers and is overexpressed in more than 90% of pancreatic cancer, which makes it an attractive target for cancer immunotherapy. As a self-protein, MUC1 shows a low immunogenicity because of immune tolerance, and the most effective approach to breaking immune tolerance is alteration of the antigen structure. In this study, the altered MUC11068-1076Y1 epitope (YLQRDISEM) by modification of amino acid residues in sequences presented a higher immunogenicity and elicited more CTLs relative to the wild-type (WT) MUC11068-1076 epitope (ELQRDISEM). In addition, the altered MUC11068-1076Y1 epitope was found to cross-recognize pancreatic cancer cells expressing WT MUC1 peptides in an HLA-A0201-restricted manner and trigger stronger immune responses against pancreatic cancer via the perforin/granzyme apoptosis pathway. As a potential HLA-A0201-restricted CTL epitope, the altered MUC11068-1076Y1 epitope is considered as a promising target for immunotherapy of pancreatic cancer. Alteration of epitope residues may be feasible to solve the problem of the low immunogenicity of TAA and break immune tolerance to induce immune responses against human cancers.

Changes of bacterial community in arable soil after short-term application of fresh manures and organic fertilizer.

The application of animal manure is highly recommended in agricultural production. However, the effect of different kinds of manures on bacterial community in farmland still remains unclear. In this study, a short-term field experiment was conducted to investigate the rapid effects of pig manure (PM), chicken manure (CM) and organic fertilizer (OF, composted by pig manure) application on soil physicochemical properties and soil bacterial community. The results showed that the application of CM and OF significantly increased soil bacterial richness (p < 0.05), mainly correlated with the increase of soil total nitrogen. Compared with CM and PM, OF had the greatest disturbance to soil bacterial structure. And total phosphorus showed the highest correlation with bacterial community. Meanwhile, the application of OF reduced the relative abundance of Actinobacteria, the organic matter synthetic bacteria, and Nitrospirae, the nitrifying bacteria, by 17.18% and 40.00%, respectively. 16S functional prediction analysis results shows that the application of OF increased the relative abundance of genes encoding Ribulose-1,5-bisphosphate carboxylase/oxyg (RuBsiCO), the genes involved in soil Calvin cycling, by 20.51%, and increased the relative abundance of genes encoding nitrous-oxide reductase by 44.86%. In conclusion, Short-term application of OF had greater disturbance to soil bacteria than CM and PM, and it had a significant influence on soil functional bacteria and genes involved in soil carbon and nitrogen cycling.

Association of exposure to PM2.5-bound metals with maternal thyroid function in early pregnancy.

Epidemiological evidence linking metals bound to ambient particulate matters with aerodynamic diameter less than 2.5 µm (PM2.5) and maternal thyroid function is limited. In this study, we investigated the association of PM2.5-bound metals with maternal thyroid hormones (TH) during the first trimester. We retrospectively reviewed data for 2528 pregnant women attending prenatal care in Jinhua Maternal and Child Health Care Hospital, Jinhua, China, from January to December 2018. Information including thyroid hormone levels and demographics was retrieved from existing medical records. We analyzed the concentration of 10 metals for collected particulate samples, and estimated their exposure levels during the first trimester for each woman. We employed multivariate linear regression models to estimate the association of exposure to individual PM2.5-bound metals with serum levels of maternal TH, and weighted quantile sum (WQS) to estimate the overall association of exposure to PM2.5-bound metals within a mixture. Higher exposures to most of the PM2.5-bound metals were associated with lower levels of maternal free thyroxine (FT4) and free triiodothyronine (FT3). The thyroid peroxidase antibody (TPOAb) or thyroglobulin antibody (TgAb) status had no effect modification on the observed associations. WQS analyses further suggested that Be, Ni, Tl and Ba contributed the most to the associations. These findings highlight the associations of exposure to PM2.5-bound metals with maternal thyroid function, and emphasize the public health significance of commitments to improve air quality.

A clinician survey for management of the secondary immunodeficiency caused by hematological malignancies in China.

ABSTRACT: Unlike Western countries, there are still few clinical immunology specialists in China, and the optimal care for secondary immunodeficiency caused by hematological malignancies is unknown. Therefore, we initiated this clinician survey study to describe the current situation of the care for malignancy patients with hypogammaglobulinemia in China.We adapted a previously published online questionnaire of current clinical practices regarding the management of secondary immunodeficiency caused by hematological malignancies and then distributed the questionnaire to 52 hematologists in China via WeChat mobile software; the survey collected demographic details, starting dosage, target immunoglobulin (Ig) level, monitoring, criteria for stopping Ig replacement, vaccination use, and oral antibiotic prophylaxis for hypogammaglobulinemia patients.Forty-eight hematologists responded. 28(58.33%) respondents had more than 10 years of experience. Nevertheless, 40(83.33%) respondents reported that they did not use any specific criteria for prophylactic Ig replacement in hypogammaglobulinemia patients. However, 27(56.25%) respondents reported that they had used intravenous immunoglobulin (IVIG); however, the starting dose, frequency, and target Ig level were significantly varied. Additionally, the criteria for stopping Ig replacement were significantly varied. Only one respondent (2.08%) used subcutaneous immunoglobulin (SCIG). Moreover, 35(72.92%) respondents reported no vaccination prior to Ig replacement, and 47(97.92%) respondents reported that they had not used antibiotic prophylaxis in secondary hypogammaglobulinemia patients.Official guideline for the care for secondary immunodeficiency (SID) of the hematological malignancies patients should be issued in China, and significant attention of the hematologists should be paid to the use of prophylactic antibiotics and Ig replacement for the care of patients with hypogammaglobulinemia caused by hematological malignancies, as these agents could significantly reduce the infection rate in China.

Residue substitution enhances the immunogenicity of neoepitopes from gastric cancers.

OBJECTIVE: Neoantigens arising from gene mutations in tumors can induce specific immune responses, and neoantigen-based immunotherapies have been tested in clinical trials. Here, we characterized the efficacy of altered neoepitopes in improving immunogenicity against gastric cancer. METHODS: Raw data of whole-exome sequencing derived from a patient with gastric cancer were analyzed using bioinformatics methods to identify neoepitopes. Neoepitopes were modified by P1Y (the first amino acid was replaced by tyrosine) and P2L (the second amino acid was replaced by leucine). T2 binding and stability assays were used to detect the affinities between the neoepitopes and the HLA molecules, as well as the stabilities of complexes. Dendritic cells (DCs) presented with neoepitopes stimulated naïve CD8+ T cells to induce specific cytotoxic T lymphocytes. ELISA and carboxyfluorescein succinimidyl ester were used to detect IFN-γ and TNF-α levels, and T cell proliferation. Perforin was detected by flow cytometry. The cytotoxicity of T cells was determined using the lactate dehydrogenase assay. RESULTS: Bioinformatics analysis, T2 binding, and stability assays indicated that residue substitution increased the affinity between neoepitopes and HLA molecules, as well as the stabilities of complexes. DCs presented with altered neoepitopes stimulated CD8+T cells to release more IFN-γ and had a greater effect on promoting proliferation than wild-type neoepitopes. CD8+T cells stimulated with altered neoepitopes killed more wild-type neoepitope-pulsed T2 cells than those stimulated with wild-type neoepitopes, by secreting more IFN-γ, TNF-α, and perforin. CONCLUSIONS: Altered neoepitopes exhibited greater immunogenicity than wild-type neoepitopes. Residue substitution could be used as a new strategy for immunotherapy to target neoantigens.

[Effects of Manure and Organic Fertilizer Application on Soil Microbial Community Diversity in Paddy Fields].

To investigate the effect of typical manure application on soil microbes in paddy fields, a field experiment on manure application in Chongming Island was carried out. The composition and variety of soil microorganisms in rice paddy fields were analyzed using high-throughput theory (CK), chicken manure (CM), pig manure (PM), and organic fertilizer (OF). The results showed that compared with CK, the application of organic manure increased soil organic matter (SOM), and the application of chicken manure significantly increased the soil ammonia nitrogen (NH4+-N) and total nitrogen (TN) contents (P<0.05). Soil microbial diversity in the PM group was significantly higher than that in the CK group (P<0.05), and the soil microbial community richness in the OF group was significantly higher than that in the CM group (P<0.05). pH, total phosphorus (TP), total nitrogen, and Pb were the important factors affecting the composition of soil microbial communities in paddy fields. The microbial community structure in the CM group was significantly different from those in the other three groups. Compared with CK, the OF group increased the relative abundance of Nitrospira, and the CM group significantly reduced the relative abundance of the denitrifying bacteria Ignavibacteriae (P<0.01) to 40.56%, but significantly increased the relative abundance of nitrifying bacteria Thauera(P<0.05) to 203.00%. The PM group significantly increased the relative abundance of ammoniated bacteria Armatimonadetes (P<0.05) to 57.51% and the anaerobic strain Anaerolinea to 102.00%. The application of chicken manure and pig manure significantly increased the relative abundance of pathogens Pseudomonas and Flavisolibacter (P<0.05), respectively, while the application of organic manure reduced the relative abundance of Flavisolibacter. Overall, the application of manure increased the abundance of bacteria involved in the nitrogen cycle of paddy soils and played a positive role in regulating the nitrogen balance in paddy soils. However, direct application of chicken manure and fresh pig manure increased the abundance of pathogenic bacteria, which had a certain degree of stress on the soil health in the paddy field.