Inhibition of ATM promotes PD-L1 expression by activating JNK/c-Jun/TNF-α signaling axis in triple-negative breast cancer.

Triple-negative breast cancer (TNBC) is a heterogeneous subtype of breast cancer. Anti-PD-1/PD-L1 treatment for advanced TNBC is still limited to PD-L1-positive patients. Ataxia telangiectasia mutated (ATM) is a switch molecule for homologous recombination and repair. In this study, we found a significant negative correlation between ATM and PD-L1 in 4 TNBC clinical specimens by single-cell RNA sequencing (scRNA-seq), which was confirmed by immunochemical staining in 86 TNBC specimens. We then established ATM knockdown TNBC stable cell lines to perform in vitro studies and animal experiments, proving the negative regulation of PD-L1 by ATM via suppression of tumor necrosis factor-alpha (TNF-α), which was confirmed by cytokine array analysis of TNBC cell line and analysis of clinical specimens. We further found that ATM inhibits TNF-α via inactivating JNK/c-Jun by scRNA-seq, Western blot and luciferase reporter assays. Finally, we identified a negative correlation between changes in phospho-ATMS1981 and PD-L1 levels in TNBC post- and pre-neoadjuvant therapy. This study reveals a novel mechanism by which ATM negatively regulates PD-L1 by downregulating JNK/c-Jun/TNF-α in TNBC, shedding light on the wide application of immune checkpoint blockade therapy for treating multi-line-resistant TNBC.

Effects of tumor-infiltrating lymphocytes on nonresponse rate of neoadjuvant chemotherapy in patients with invasive breast cancer.

High level of tumor-infiltrating lymphocytes (TILs) can predict the rate of total pathological complete remission (tpCR) of breast cancer patients who receive neoadjuvant chemotherapy (NACT). This study focused on evaluating the data of patients whose primary tumor and/or lymph node metastasis show nonresponse (NR) to NACT, trying to provide a basis for the clinical decision which patients will develop NACT resistance. The study included breast cancers from 991 patients who received NACT. ROC curve analysis confirmed that TILs showed significant predictive value for NR of hormone receptor (HR)+HER2- and triple-negative breast cancer (TNBC). Among HR+HER2- breast cancer, TILs ≥ 10% was an independent predictor for low NR rate. Furthermore, positive correlation of TILs with Ki67 index and Miller-Payne grade, and negative correlation with ER and PR H-scores were only identified in this subgroup. In TNBC, TILs ≥ 17.5% was an independent predictor for low NR rate. The predictive value of low TILs on NR may facilitate to screen patients with HR+HER2- or TNBC who may not benefit from NACT. HR+HER2- breast cancer with low levels of TILs should be carefully treated with neoadjuvant chemotherapy, and other alternatives such as neoadjuvant endocrine therapy can be considered.

A novel variant of SLC4A1 for hereditary spherocytosis in a Chinese family: a case report and systematic review.

BACKGROUND: The incidence of hereditary spherocytosis (HS) is approximately 1:2000 in the western population, while it is much lower in the Chinese population. It is difficult to make a definite diagnosis due to the variable genotypic features and the lack of well-documented evidence for HS patients. Gene sequence examination is helpful for clear diagnosis. CASE PRESENTATION: We presented the case of a 29-year-old male HS patient with skin yellowness, anorexia, and cholecystolithiasis as the first manifestations. Laboratory examination of the patient and his parents showed a mild reduction in hemoglobin and mean corpuscular hemoglobin concentration, increased reticulocytes, and promotion of indirect bilirubin in the patient and his father. Furthermore, small globular red blood cells with increased osmotic fragility were observed. In particular, the eosin-5'-maleimide binding test provided the strong evidence that band 3 protein was deleted in the erythrocyte membrane. Next-generation sequencing (NGS) and Sanger sequencing further demonstrated a heterozygous nonsense variant (exon16, c.G1985A: p.W662X) in SLC4A1, inherited from his father. Thus, the patient was diagnosed with HS, and then was effectively treated. After splenectomy, the anemia was relieved without any obvious unpleasant side effects. CONCLUSION: We report an extremely rare case of HS in China that presented with hereditary hemolytic anemia with band 3 deletion resulting from a novel variant of SLC4A1, and systematically review a large number of related literatures. This study, therefore, significantly contributes to the literature on HS.

STAT3 and PD-L1 are negatively correlated with ATM and have impact on the prognosis of triple-negative breast cancer patients with low ATM expression.

INTRODUCTION: Triple-negative breast cancer (TNBC) is known for its aggressive behaviors and lacking of effective treatment. Programmed cell death ligand-1 (PD-L1) inhibitor has just been approved for using in the management of advanced TNBC. To accurately screen TNBC sensitive to anti-PD-L1 treatment and to explore the feasibility of the ataxia-telangiectasia mutation protein (ATM) inhibitor combined with PD-L1 inhibitor, radiotherapy and chemotherapy, we focus on whether ATM participates in the regulation of PD-L1 and affects the prognosis of patients through c-Src, signal transducer and activator of transcription 1&3 (STAT1 and STAT3). MATERIALS AND METHODS: We used immunohistochemical staining to explore the relationship of ATM with c-Src, STAT1, STAT3, PD-1/PD-L1, Tumor-infiltrating lymphocytes (TILs), as well as other clinicopathologic features in 86 pathological stage III TNBCs. Their impact on prognosis was also explored. RESULTS: We found ATM expression was negatively correlated with STAT1, STAT3, PD-L1, TILs and CD8 + cells in TNBC. STAT1 positively correlated the expression of PD-L1. In TNBC with ATM low expression, STAT3 was an independent factor for improved prognosis, while PD-L1 was an independent negative prognostic factor. Furthermore, in low ATM group, the phosphorylation of tyrosine at position 419 of c-Src (p-c-src Y419) was correlated with the overexpression of STAT3. CONCLUSION: Locally advanced TNBC with low ATM expression may be more likely to benefit from anti-PD-L1 inhibitors. The feasibility of ATM functional inhibitor combined with immune checkpoint blockade therapies in the treatment of TNBC is also worthy of further exploration. Our study suggests that STAT3 has different impacts on tumor progression in different tumors.

CMTM Family and Gastrointestinal Tract Cancers: A Comprehensive Review.

Gastrointestinal tract cancers are a highly heterogeneous group of malignant diseases, contributing significantly to the burden of death worldwide. Chemokine-like factor (CKLF)-like MARVEL transmembrane domain-containing family (CMTMs) plays important roles in cancer development and progression. Since the first member was cloned, there have been abundant studies on the relationships between the CMTM family and human cancers. It has been reported that the CMTM family has a large potential prognostic value for multiple cancers. Meanwhile, upregulated or downregulated expression of the family members was related to advanced tumor stage, metastasis, and overall survival. Studies have also reported that these proteins play critical roles in antitumor immunity. We performed a systematic review to sum up the latest advances of CMTM family' roles in gastrointestinal tract cancers, with a primary focus on hepatocellular carcinoma and gastric carcinoma.

The diagnostic value of interleukin 6 as a biomarker for gastric cancer: A meta-analysis and systematic review.

BACKGROUND: Gastric cancer is one of the most common cancers and a main cause of global cancer death. The expression of interleukin 6 is associated with the risk of gastric cancer. But the diagnostic accuracy of interleukin 6 remains unclear. This study was designed to assess the diagnostic performance of interleukin 6 in gastric cancer diagnosis. METHODS: The related data was obtained from Oncomine and studied using bioinformatics analysis. The PubMed, Embase, Cochrane Library, Web of science databases were searched for related studies published from inception to July 14, 2020. Measuring tools of diagnostic performance including sensitivity, specificity, and diagnostic odds ratio were pooled using bivariate mixed-effects meta-analysis model. The summery receiver operator characteristic curves were plotted. RESULTS: The result from Oncomine showed that the expression of interleukin 6 in gastric cancer (GC) patients was higher than the normal groups (P < .05). Furthermore, a total of 4 eligible articles were enrolled, containing 390 cases and 404 controls. The diagnostic results were as follows: a sensitivity of 0.80 (95% confidence interval [CI] 0.57-0.92), a specificity of 0.86 (95% CI 0.74-0.93), a positive likelihood ratio of 5.76 (95% CI 3.49-9.49), a negative likelihood ratio of 0.23 (95% CI 0.11-0.51) and a diagnostic odds ratio of 24.58 (95% CI 14.14-42.73). The summary area under the receiver operating characteristic curves was 0.90 (95% CI 0.87-0.93). CONCLUSION: Higher interleukin 6 expression was detected in GC patients, and interleukin 6 could be a helpful indicator of diagnosis of gastric cancer. Further large-scale prospective studies are required for identifying the diagnostic value of interleukin 6 in gastric cancer.

The Cd phytoextraction potential of hyperaccumulator Sedum alfredii-oilseed rape intercropping system under different soil types and comprehensive benefits evaluation under field conditions.

Soil trace elements (TEs) contamination has become a worldwide problem in arable lands and poses great risk to human health via food chain. Intercropping of hyperaccumulator and cash crops is now proposed as a promising alternative phytoremediation technique to address the issue. However, the effect of intercropping in different soil types and field-scale benefits evaluation are rarely reported. A greenhouse pot experiment and a field trial were therefore designed to explore the effects of intercropping Sedum alfredii (hyperaccumulative population) and oilseed rape on Cd phytoextraction potential, Cd transport and crop production, as well as establishing a feasible assessment framework on the basis of benefits evaluation in contaminated soils. Compared with oilseed rape monoculture, intercropping with S. alfredii significantly and consistently increased biomass, seed yield and Cd accumulation in oilseed rape in five typical soil types. Accumulations of Cd varied with soil types, ranging from 22.8 to 4000 µg pot-1. Stepwise multiple linear regression analysis (SMLRA) showed Cd concentrations in plants were related to available phosphorus (AP), pH, soil organic matter (OM), available potassium (AK), silt and sand; R2 values varied from 0.834 to 0.994 (P < 0.05). A field trial also verified that intercropping could significantly enhance Cd phytoextraction. The highest index for comprehensive benefits evaluation was 0.61 observed in the S. alfredii and oilseed rape intercropping system. This system presented higher Cd phytoextraction potential and comprehensive benefits index whilst allowing ongoing agricultural activities in slightly and moderately Cd-contaminated soils. These results provide a possible technical approach for phytoremediation practice and give new insights into theoretical reference for development of Cd phytoextraction and benefits evaluation.

Dual PI3K/mTOR inhibitor NVP­BEZ235 decreases the proliferation of doxorubicin­resistant K562 cells.

Acute myelogenous leukemia (AML) is frequently accompanied by a poor prognosis. The majority of patients with AML will experience recurrence due to multiple drug resistance. Our previous study reported that targeting the mTOR pathway may increase cell sensitivity to doxorubicin (Doxo) and provide an improved therapeutic approach to leukemia. However, the effect and mechanism of action of NVP­BEZ235 (BEZ235), a dual inhibitor of PI3K/mTOR, on Doxo­resistant K562 cells (K562/A) is yet to be elucidated. Therefore, the aim of the present study was to investigate the effects of BEZ235 on K562/A cell proliferation. K562/A cells was investigated using CCK­8, flow cytometry and western blotting, following BEZ235 treatment. It was observed that BEZ235 significantly decreased the viability of K562/A cells. In addition, BEZ235 arrested K562/A cells at the G0/G1 phase, and reduced the protein expression levels of CDK4, CDK6 and cyclin D1. Apoptotic cells were more frequently detected in K562/A cells treated with BEZ235 compared with the control group (12.97±0.91% vs. 7.37±0.42%, respectively; P<0.05). Cells treated with BEZ235 exhibited downregulation of Bcl­2 and upregulation of Bax. Furthermore, BEZ235 treatment markedly decreased the activation of the PI3K/AKT/mTOR pathway and its downstream effectors. Thus, these results demonstrated that BEZ235 inhibited cell viability, induced G0/G1 arrest and increased apoptosis in K562/A cells, suggesting that BEZ235 may reverse Doxo resistance in leukemia cells. Therefore, targeting the PI3K/mTOR pathway may be of value as a novel therapeutic approach to leukemia.

Trefoil factor 1 and gastrokine 2 inhibit Helicobacter pylori-induced proliferation and inflammation in gastric cardia and distal carcinogenesis.

Helicobacter pylori (H. pylori) infection has been associated with non-cardia adenocarcinoma in the stomach, while its role in gastric cardia adenocarcinoma (GCA) remains controversial. In addition, the association between H. pylori and the protective factors trefoil factor 1 (TFF1) and gastrokine 2 (GKN2) in gastroesophageal adenocarcinomas has not been fully investigated. Therefore, the mRNA and protein expression levels of TFF1 and GKN2 in GCA and distal gastric adenocarcinoma (DGA) were analyzed using quantitative PCR (qPCR) and immunohistochemistry, and the association with H. pylori infection was investigated. In addition, the effects of TFF1 and GKN2 overexpression on H. pylori-induced cells were investigated using western blot and reverse transcription-qPCR analysis. The comparative analysis of 16S rRNA-positive mRNA expression between GCA and DGA showed no statistically significant difference. However, the rate of the H. pylori vacuolating toxin A (VacA) genotype was significantly higher in GCA (49.2%) compared with that in DGA (26.9%; P<0.05). H. pylori infection downregulated the mRNA and protein expression levels of TFF1 and GKN2 in gastric tumor tissues, and the mRNA expression level of TFF1 and GKN2 was also markedly decreased in vitro. Furthermore, the cell proliferation varied in H. pylori total protein treatment group with the different doses. Notably, treatment with 20 µg/ml H. pylori total protein for 24 h resulted in the highest cellular proliferation rate. In addition, TFF1 and GKN2 overexpression inversely inhibited H. pylori-induced cell proliferation and upregulated NF-κB, tumor necrosis factor-α, IL-1ß, IL-2, IL-4 and IL-6. The results of the present study indicate that H. pylori, particularly the VacA+ strain, plays an important role in GCA pathogenesis in high-risk areas of China, while TFF1/GKN2 inhibits H. pylori-induced cell proliferation and inflammation in GCA and DGA.

Pulmonary tuberculosis presenting as henoch-schönlein purpura: Case report and literature review.

INTRODUCTION: Henoch-Schönlein purpura (HSP) is an extremely rare condition in patients with pulmonary tuberculosis, with only a few reported cases. Compared to patients with typical clinical symptoms, it is difficult to make a definitive diagnosis when HSP presents as an initial manifestation in pulmonary tuberculosis patients. Herein, a case of pulmonary tuberculosis that showed HSP at first was reported, and the related literatures were reviewed. PATIENT CONCERNS: A 24-year-old man presented with palpable purpura on the extremities, accompanied by abdominal pain, bloody stools, and knee pain. DIAGNOSES: The patient was diagnosed with pulmonary tuberculosis based on the results of interferon gamma release assays, purified protein derivative test, and computed tomography. INTERVENTIONS: The patient was treated with vitamin C and chlorpheniramine for 2 weeks, and the above-mentioned symptoms were relieved. However, 3 weeks later, the purpura recurred with high-grade fever and chest pain during the inspiratory phase. The patient was then treated with anti-tuberculosis drugs, and the purpura as well as the high fever disappeared. OUTCOMES: The patient recovered well and remained free of symptoms during the follow-up examination. CONCLUSION: Pulmonary tuberculosis presenting with HSP as an initial manifestation is not common. Therefore, it is difficult to clinically diagnose and treat this disease. When an adult patient shows HSP, it is important to consider the possibility of tuberculosis to avoid misdiagnosis and delayed treatment.

Responses of soil bacterial community and Cd phytoextraction to a Sedum alfredii-oilseed rape (Brassica napus L. and Brassica juncea L.) intercropping system.

Soil pollution with heavy metals has become a common problem in agricultural ecosystems and poses a threat to food safety and human health. Intercropping is now considered a promising alternative to address this issue. However, our understandings about the influences of intercropping systems on rhizosphere microbiota composition and their association with plant performance are still limited. In this study, rhizobox microcosm experiments were conducted to investigate the influence of cropping regimes (i.e. monoculture and intercropping) on the rhizosphere bacterial microbiota and their linkages with the phytoextraction of cadmium (Cd) by Zhongyouza 19 (Brassica napus L.), Xikou Huazi (Brassica juncea L.) and Sedum alfredii using 16S rRNA gene sequencing. Cadmium accumulation in shoots of B. napus and B. juncea grown under intercropping were enhanced by 370% and 27.8% respectively, as compared to monoculture. Soil compartmentation as a major determinant explained 57.6% of the rhizosphere bacterial microbiota variation, whereas plant species and cropping regime accounted for 26.4% of the variation. The overall abundance of the taxa was Proteobacteria, Acidobacteria, Bacteroidetes, Chloroflexi, Verrucomicrobia, and Actinobacteria. Intercropping significantly enriched amplicon sequence variants (ASVs) abundance belonging to Actinobacteria, Bacilli, Deltaproteobacteria while depleting that of Acidobacteria in rhizosphere. Intercropping with S. alfredii influenced more on microbial composition of B. napus rhizosphere. The change in rhizosphere bacterial communities was related to metal availability, soil properties, and plant parameters. The enriched families of Pedosphaeraceae, Ruminococcaceae, Chitinophagaceae, Gemmatimonadaceae, Nitrosomonadaceae, and Parachlamydiaceae were positively correlated with metal concentration in plants. These results indicate that S. alfredii and oilseed rape intercropping could be a promising approach for enhancing the remediation of Cd contaminated soil. Understanding the complex plant-microbe-metal interactions of intercropping system could facilitate the development of remediation strategy for phytoremediation of contaminated soils and sustainable agricultural production.

Accumulation and distribution of cadmium and lead in 28 oilseed rape cultivars grown in a contaminated field.

Heavy metal pollution in soils has become an important concern for human health. Therefore, it is vital to develop suitable remediation strategies for contaminated soils. Oilseed rape tolerates high concentrations of heavy metals and is a promising candidate for the phytoextraction of cadmium (Cd) and lead (Pb) from metal-contaminated soils. A field experiment was conducted to evaluate 28 oilseed rape cultivars including Brassica napus L. and Brassica juncea L. for their ability to accumulate Cd and Pb. These cultivars were grown in a field co-contaminated with Cd (0.78 mg kg-1) and Pb (330 mg kg-1). The results showed that concentrations in shoots ranged from 1.22 to 3.01 mg kg-1 for Cd and from 10.8 to 29.5 mg kg-1 for Pb. Cadmium and Pb accumulations in shoots could reach 83.4 and 799 µg plant-1, respectively. The majority of translocation factors (TFs) for Cd (> 1.0) were higher than for Pb (≤ 1.0). However, concentrations of Cd and Pb in seeds were much lower, in the range of 0.04 to 0.21 mg kg-1 and 0.04 to 0.51 mg kg-1, respectively. The seed yields of oilseed rape varied from 1238 to 2904 kg ha-1, with a mean value of 2289 kg ha-1. Among the cultivars, three (OS-9, OS-12, and OS-15) were selected as Cd and Pb potential accumulators, with Cd accumulation in shoots being 2.74-3.70 times higher and Pb accumulation in shoots being 3.37-5.23 times higher as compared with the lowest accumulating cultivar. These selected cultivars (B. napus) have application potential for phytoextraction of Cd and Pb from polluted soils without stopping agricultural activities and accompanying food safety issues.

Distribution, availability and translocation of heavy metals in soil-oilseed rape (Brassica napus L.) system related to soil properties.

Heavy metals contamination in agricultural soil has become a worldwide problem, and soil characteristics modulate metal availability in soils. Four field experiments were conducted simultaneously to evaluate concentration and distribution of cadmium (Cd) and lead (Pb) in 39 oilseed rape cultivars at four agricultural locations with different contamination levels of Cd and Pb, as well as the influence of soil characteristics together with soil total and bioavailable Cd and Pb concentration on metal transfer from soil to oilseed rape. Shoot concentrations of Cd and Pb in oilseed rape cultivars ranged from 0.09 to 3.18 and from 0.01 to 10.5 mg kg-1 across four sites. For most cultivars, Cd concentration in root or shoot were higher than pod and lowest in seed, while the highest Pb concentration was observed in root followed by shoot and seed. Stepwise multiple linear regression analysis allows for a better estimation of Cd and Pb concentration in oilseed rape while taking soil properties into consideration. The results demonstrated that Cd and Pb concentration in oilseed rape were correlated with soil organic matter (OM), cation exchange capacity (CEC), available phosphorus (AP), available potassium (AK), sand, soil total and available Cd and Pb concentration, and R2 varied from 0.993 to 0.999 (P < 0.05). The Cd and Pb levels found in oilseed rape indicated its phytoextraction potential for Cd and Pb co-contaminated agricultural soils in winter without stopping agricultural activities.

Immobilization of cadmium and lead in contaminated paddy field using inorganic and organic additives.

Heavy metal contamination of agricultural soils has posed a risk to environment and human health. The present study was conducted to assess the effectiveness of soil amendments for reducing cadmium (Cd) and lead (Pb) uptake by rice (Oryza sativa L) in a contaminated field. The soil amendments used include lime, DaSan Yuan (DASY), DiKang No.1 (DEK1), biochar, Fe-biochar, Yirang, phosphorus fertilizer, (Green Stabilizing Agent) GSA-1, GSA-2, GSA-3, and GSA-4, applied at 1% rate in a field experiment. The results exposed that GSA-4 treatment showed best effects on reducing Cd and Pb phytoavailability in soil and uptake by early rice. Linear increase in pH (i.e. 5.69 to 6.75) was recorded in GSA-4 amended soil from sowing to the 3rd month of growth season. GSA-4 decreased DTPA extractable contents of cadmium (Cd) from 0.324 to 0.136 mg kg-1 soil and lead (Pb) from 53.21 to 24.68 mg kg-1 soil at 90 days of amendment. Treatment with GSA-4 improved rice growth (56%) and grains yield (42%). The enhancement effects on grain yield may be result from the positive effects of GSA-4 application on increasing photosynthesis (116%) and transpiration rate (152%) as compared to the control. Significant reduction in Cd and Pb uptake in shoot (42% and 44%) and in grains (77 and 88%), was observed, respectively in GSA-4 treatment as compared with the control. Moreover, negative correlation was recorded between DTPA extractable Cd/Pb and soil pH that directly depended on applied amendments. In short, use of combined amendment (GSA-4) was more effective for immobilizing heavy metals in contaminated paddy field, and secures rice safe production, as compared other tested amendment products.