Leaching behavior and release mechanism of pollutants from different depths in a phosphogypsum stockpile.

Phosphogypsum (PG), a byproduct during the production of phosphoric acid and phosphate fertilizers, is predominantly stockpiled with a height greater than hundreds of meters. In this study, the leaching behavior of pollutants from PG stored at different depths was systematically investigated through batch tests, column tests, and geochemical modeling. PG samples were collected at different depths within a range of 48 m from a large-scale PG stack in China. The results showed that the pH, electrical conductivity, and elemental concentration of the leachate exhibited spatial variability in terms of the depth distribution, with evident bottom enrichment effects for metals and soluble salts. The pH-dependent leaching tests investigated the impact of pH variations on the solubility of various elements in PG, with a specific focus on elements precipitation occurring within the natural pH range. The geochemical modeling of leaching tests conducted by PHREEQC enabled the identification of the dominant phases controlling the solubilization of the elements, as well as the dynamic process of changes in element forms and concentrations with pH variation. Column leaching tests reveal the differences in pollutant properties between the unsaturated and saturated zones within the PG stack and categorize the leaching mechanisms of elements into three models including dissolution, diffusion, and wash-off. This study aims to reveal the leaching characteristics of PG at different depths, so as to provide a data foundation for the design of liner system, leachate management strategies, and remediation of heavy metal pollution of PG stack sites.

[Corrigendum] miR­146b­5p inhibits tumorigenesis and metastasis of gallbladder cancer by targeting Toll­like receptor 4 via the nuclear factor­κB pathway.

Following the publication of the article, the authors drew to the Editor's attention that, in Fig. 4D on p. 7, the data correctly shown to represent the E­cadherin bands for the "NOZ" experiment had inadvertently been used to show the Vimentin bands. However, the authors retained their original data, and the corrected version of Fig. 4, now showing the correct data for the Vimentin bands in Fig. 4D for the "NOZ" experiment, is shown on the next page. Note that this error did not grossly affect either the results or the conclusions reported in this work. All the authors agree with the publication of this Corrigendum, and are grateful to the Editor of Oncology Reports for granting them the opportunity to correct the error that was made during the assembly of this figure. Lastly, the authors apologize to the readership for any inconvenience this error may have caused. [Oncology Reports  45: 15, 2021; DOI: 10.3892/or.2021.7966].

Targeting the chromatin structural changes of antitumor immunity.

Epigenomic imbalance drives abnormal transcriptional processes, promoting the onset and progression of cancer. Although defective gene regulation generally affects carcinogenesis and tumor suppression networks, tumor immunogenicity and immune cells involved in antitumor responses may also be affected by epigenomic changes, which may have significant implications for the development and application of epigenetic therapy, cancer immunotherapy, and their combinations. Herein, we focus on the impact of epigenetic regulation on tumor immune cell function and the role of key abnormal epigenetic processes, DNA methylation, histone post-translational modification, and chromatin structure in tumor immunogenicity, and introduce these epigenetic research methods. We emphasize the value of small-molecule inhibitors of epigenetic modulators in enhancing antitumor immune responses and discuss the challenges of developing treatment plans that combine epigenetic therapy and immunotherapy through the complex interaction between cancer epigenetics and cancer immunology.

Periodic injection of liquefied kitchen and food waste in municipal solid waste: Effects on leachate and gas generation.

With continuous advancements in the zero-waste strategy in China, transportation of fresh municipal solid waste to landfills has ceased in most first-tier cities. Consequently, the production of landfill gas has sharply declined because the supply of organic matter has decreased, rendering power generation facilities idle. However, by incorporating liquefied kitchen and food waste (LKFW), sustainable methane production can be achieved while consuming organic wastewater. In this study, LKFW and water (as a control group) were periodically injected into high and low organic wastes, respectively. The biochemical characteristics of the resulting gas and leachate were analyzed. LKFW used in this research generated 19.5-37.6 L of methane per liter in the post-methane production phase, highlighting the effectiveness of LKFW injection in enhancing the methane-producing capacity of the system. The release of H2S was prominent during both the rapid and post-methane production phases, whereas that of NH3 was prominent in the post-methane production phase. As injection continued, the concentrations of chemical oxygen demand, 5-d biological oxygen demand, total organic carbon, ammonia nitrogen, total nitrogen, and oil in the output leachate decreased and eventually reached levels comparable to those in the water injection cases. After nine rounds of injections, the biologically degradable matter of the two LKFW-injected wastes decreased by 8.2 % and 15.1 %, respectively. This study sheds light on determining the organic load, controlling odor, and assessing the biochemical characteristics of leachate during LKFW injection.

A swift expanding trend of extracellular vesicles in spinal cord injury research: a bibliometric analysis.

Extracellular vesicles (EVs) in the field of spinal cord injury (SCI) have garnered significant attention for their potential applications in diagnosis and therapy. However, no bibliometric assessment has been conducted to evaluate the scientific progress in this area. A search of articles in Web of Science (WoS) from January 1, 1991, to May 1, 2023, yielded 359 papers that were analyzed using various online analysis tools. These articles have been cited 10,842 times with 30.2 times per paper. The number of publications experienced explosive growth starting in 2015. China and the United States led this research initiative. Keywords were divided into 3 clusters, including "Pathophysiology of SCI", "Bioactive components of EVs", and "Therapeutic effects of EVs in SCI". By integrating the average appearing year (AAY) of keywords in VoSviewer with the time zone map of the Citation Explosion in CiteSpace, the focal point of research has undergone a transformative shift. The emphasis has moved away from pathophysiological factors such as "axon", "vesicle", and "glial cell" to more mechanistic and applied domains such as "activation", "pathways", "hydrogels" and "therapy". In conclusions, institutions are expected to allocate more resources towards EVs-loaded hydrogel therapy and the utilization of innovative materials for injury mitigation.

The complex impact of family caregiver kinship type on the rehabilitation of patients with mental disorders in rural communities of China.

BACKGROUND: In 2009, the Chinese government incorporated severe mental disorders into the central subsidized local health funding project for the effective management, treatment, and reintegration of patients with severe mental disorders from hospitals into the community (Project 686). The conditions that were classified as 'severe' by this project included: schizophrenia, schizoaffective disorders, paranoid disorders, bipolar disorders, mental disorders caused by epilepsy, and mental retardation accompanying mental disorders. Patients in rural communities received better care after project implementation, of which 62.91% were farmers. OBJECTIVE: This paper attempts to investigate the complex impact of Project 686 on the levels of rehabilitation of patients by their families. METHODS: The last follow-up visit of the community psychiatrists in city H in 2020 was used as the time point. Finally, 174 samples were used in the analysis model. The type of kinship between family caregivers and patients with mental disorders was operationalized according to the information provided under the 'primary caregiver' item within the basic information section of the follow-up form. The software Stata15 was used to perform descriptive statistics, baseline regression model analysis, and a robustness test on the types of kinship identified and patients' recovery. RESULTS: The types of kinship, current symptoms, and medication use all affected patients' recovery, were found to have regression coefficients of -0.148, -1.756, and 0.902, respectively. Parents of patients with mental disorders remain in the caregiver category with the largest proportion. Community acceptance of patients is high; current symptoms, medication use, and types of caregiver-patient relationships influence patients' recovery levels. CONCLUSIONS AND DISCUSSION: Project 686 has solved some of the rehabilitation and living difficulties of patients with mental disorders in rural communities. The types of kinship between family caregivers and patients with mental disorders in rural communities affect patients' rehabilitation levels. Patients' current symptoms and medication use can effectively moderate the impact of kinship type on patients' recovery in terms of complete self-knowledge, productive work, and life and good social relationships. It is necessary for mental illness prevention-treatment organizations to establish supplemental, replacement, and substitution mechanisms for the life and rehabilitation of patients with mental disorders in rural communities. Furthermore, the sense of reward and concern for family caregivers should be actively enhanced, and greater scientific use of the rehabilitation function of the 'family care + village doctor management' model should be made.

LncRNA OTUD6B-AS1 overexpression promoted GPX4-mediated ferroptosis to suppress radioresistance in colorectal cancer.

BACKGROUND: Radiotherapy is widely employed in colorectal cancer (CRC) treatment but is often compromised by developed radioresistance. This study explored the mechanism of long non-coding RNA ovarian tumor domain containing 6B-antisense RNA1 (lncRNA OTUD6B-AS1) in CRC radioresistance through tripartite motif 16 (TRIM16). METHODS: CRC and non-cancerous tissues were collected and radioresistant CRC cells were established, with real-time quantitative polymerase chain reaction to determine gene expression in tissues and cells. Radioresistance was evaluated by cell counting kit-8 assay and immunofluorescence (γ-H2AX) and ferroptosis was tested by Western blot assay (ACSL4/GPX4) and assay kits (Fe2+/ROS/MDA/GSH). The association between ferroptosis and lncRNA OTUD6B-AS1-inhibited radioresistance was testified using ferroptosis inhibitor. The subcellular localization of lncRNA OTUD6B-AS1 was tested by the nuclear/cytoplasmic fractionation assay, with RNA immunoprecipitation assay to validate gene interactions. Rescue experiments were conducted to analyze the role of TRIM16 in CRC radioresistance. RESULTS: LncRNA OTUD6B-AS1 and TRIM16 were poorly expressed (P < 0.01) in CRC tissues and cells and further decreased (P < 0.01) in radioresistant CRC cells. OTUD6B-AS1 overexpression decreased cell survival (P < 0.01), increased γ-H2AX levels (P < 0.01), and elevated ferroptosis and oxidative stress (P < 0.01) after X-ray radiation. Ferroptosis inhibitor attenuated radioresistance (P < 0.01) caused by lncRNA OTUD6B-AS1 overexpression. LncRNA OTUD6B-AS1 stabilized TRIM16 mRNA via binding to HuR. TRIM16 knockdown reduced ferroptosis and increased radioresistance (P < 0.05). CONCLUSION: OTUD6B-AS1 overexpression stabilized TRIM16 via binding to HuR and increased GPX4-mediated ferroptosis, thus attenuating CRC radioresistance. Our study provided a new rationale for the treatment of CRC.

Hesperetin attenuates sepsis-induced intestinal barrier injury by regulating neutrophil extracellular trap formation via the ROS/autophagy signaling pathway.

Background: Hesperetin (HES), one of the major flavonoids that has various biological activities, such as anti-inflammatory and antioxidant activities, may preserve the intestinal barrier during sepsis. However, the detailed mechanism remains unclear. Our previous studies confirmed that neutrophil extracellular traps (NETs) may jeopardize the intestinal barrier via a reactive oxygen species (ROS)-dependent pathway during sepsis. Therefore, we hypothesized that HES may inhibit NET formation and protect the intestinal barrier function during sepsis. Methods: Mice were pretreated with HES (50 mg kg-1) intraperitoneally for one week, and sepsis models were then induced using lipopolysaccharides (LPS) (10 mg kg-1). The mice were randomly divided into three groups: (1) sham group; (2) LPS group; and (3) HES + LPS group. Twenty-four hours after LPS injection, the serum and terminal ileum specimens were collected for subsequent studies. To detect ROS production and NET formation in vitro, human neutrophils were collected and incubated with phorbol-12-myristate-13-acetate (PMA) and various concentrations of HES. The level of autophagy was measured by an immunofluorescence assay and western blot analysis. TUNEL staining was utilized to analyze cell apoptosis. Results: The outcomes demonstrated that HES decreased inflammatory cytokine and myeloperoxidase (MPO) levels in serum and attenuated distant organ dysfunction in LPS-induced septic mice. Meanwhile, HES treatment reversed intestinal histopathological damage in septic mice, improving intestinal permeability and enhancing tight junction expression. Moreover, we found that neutrophil infiltration and NET formation in the intestine were suppressed during sepsis after HES pretreatment. In vitro, HES treatment reduced PMA-induced ROS production and NET formation, which were reversed by hydrogen peroxide (H2O2) administration. Notably, HES also inhibited NET formation by reducing the microtubule-associated protein light chain 3 (LC3)-II/LC3-I ratio (an indicator of autophagy) in PMA-induced neutrophils, which was reversed by rapamycin. Moreover, when autophagy was suppressed by chloroquine or induced by rapamycin, apoptosis in cells will be switched with autophagy. Conclusion: Taken together, these findings suggest that HES may inhibit NET formation in a ROS/autophagy-dependent manner and switch neutrophil death from NETosis to apoptosis, which reduced NETs-related intestinal barrier damage, providing a novel protective role in intestinal barrier dysfunction during sepsis.

Comparative Study on Growth Index and Nutritional Quality of Female Chinese Mitten Crab Eriocheir sinensis Selected at Different Growth Periods in Rice-Crab Culture Systems.

Research was conducted on the growth performance and nutritional quality of Chinese mitten crabs (Eriocheir sinensis) during a 62-day growing period in a symbiotic coculture comprising rice and crab. Culture experiments were conducted in three rice fields of equal size (996 m2). On days 0 (July 15, D0), 15 (July 30, D15), 31 (August 15, D31), 46 (August 30, D46), and 62 (September 2, D62), tissue samples of 50 female E. sinensis were collected randomly from each rice field. The results showed that the serum growth hormone (GH) content and muscle ecdysone receptor (EcR) mRNA expression levels were higher in the D31 and D46 groups; the content of serum 20-hydroxyecdysone (20-HE) and the mRNA expression levels of retinoid X receptor (RXR), insulin-like growth factor 2 (IGF2), and chitinase (CHI) reached the maximum in the D31 group. Muscle crude protein content gradually increased; hepatopancreas crude protein and crude lipid content began to decrease after reaching the maximum value in the D0 and D15 groups, respectively; the contents of crude protein and crude lipid in the ovary significantly increased in the D46 and D62 groups (P < 0.05). The content of muscle essential amino acids (EAA) reached the maximum in the D46 group; the hepatopancreas EAA content began to decrease significantly in the D31 group (P < 0.05); and the EAA content of the ovary decreased significantly after reaching the maximum value in the D46 group (P < 0.05). The muscle contents of eicosapentaenoic acid (EPA), docosapentaenoic acid (DPA), docosahexaenoic acid (DHA), polyunsaturated fatty acids (PUFA), and n-3 polyunsaturated fatty acids (n-3PUFA) and the ratio of n-3 polyunsaturated fatty acids/n-6 polyunsaturated fatty acids (n3/n6) decreased significantly in the D31 group (P < 0.05); the hepatopancreas contents of EPA, PUFA, n-3PUFA, and n-6 polyunsaturated fatty acids (n-6PUFA) and the ratio of n3/n6 began to decrease after reaching the maximum value in the D31 group, ethyl behenate (21:0), tetracosanoic acid (24:0), DPA, and DHA contents were detected for the first time in the D31 group; the ovary PUFA, n-3PUFA contents, and n3/n6 ratio of the D46 and D62 groups were significantly lower than those of the D31 group (P < 0.05). During the experimental conditions described here, female E. sinensis raised in rice fields reached rapid growth from August 15 to August 30. Additionally, the nutritional quality of the female E. sinensis edible tissues (muscle, hepatopancreas, and ovary) began to decline after August 15, when sufficient nutrients such as protein, lipid, EAA, and PUFA should be provided to the female E. sinensis.

Comparison of the clinical efficacy of laparoscopic extralevator abdominoperineal excision (ELAPE) and non-ELAPE for low rectal cancer.

This study was performed to retrospectively analyze and compare the related clinical indicators between extralevator abdominoperineal excision (ELAPE) and non-ELAPE under laparoscopic for low rectal cancer. From June 2018 to September 2021, a total of 80 patients with low rectal cancer who underwent either of the above two types of surgeries at our Hospital were enrolled. Patients were divided into the ELAPE group and non-ELAPE group based on the different surgical methods. Preoperative general indicators, intraoperative indicators, postoperative complications, positive circumferential resection margin rate, local recurrence rate, hospital stay length, hospital expenses, and other related indicators were compared between the two groups. There were no significant differences in the comparison of preoperative indexes between the ELAPE group and non-ELAPE group, including age, preoperative BMI, and gender. Similarly, there were no significant differences in abdominal operation time, total operation time, and the number of intraoperative lymph nodes dissected between the two groups. However, the perineal operation time, intraoperative blood loss, intraoperative perforation rate, and positive circumferential resection margin rate were significantly different between the two groups. In the comparison of postoperative indexes, perineal complications, postoperative hospital stay length, and IPSS score were significantly different between the two groups. The use of ELAPE in treating T3-4NxM0 phase low rectal cancer was superior to non-ELAPE in reducing intraoperative perforation rate, positive circumferential resection margin rate, local recurrence rate, etc.

The value of a risk model combining specific risk factors for predicting postoperative severe morbidity in biliary tract cancer.

Purpose: Several surgical risk models are widely utilized in general surgery to predict postoperative morbidity. However, no studies have been undertaken to examine the predictive efficacy of these models in biliary tract cancer patients, and other perioperative variables can also influence morbidity. As a result, the study's goal was to examine these models alone, as well as risk models combined with disease-specific factors, in predicting severe complications. Methods: A retrospective study of 129 patients was carried out. Data on demographics, surgery, and outcomes were gathered. These model equations were used to determine the morbidity risks. Severe morbidity was defined as the complication comprehensive index ≥ 40. Results: Severe morbidity was observed in 25% (32/129) patients. Multivariate analysis demonstrated that four parameters [comprehensive risk score ≥1, T stage, albumin decrease value, and international normalized ratio (INR)] had a significant influence on the probability of major complications. The area under the curve (AUC) of combining the four parameters was assessed as having strong predictive value and was superior to the Estimation of Physiologic Ability and Surgical Stress System (E-PASS) alone (the AUC value was 0.858 vs. 0.724, p = 0.0375). The AUC for the modified E-PASS (mE-PASS) and Physiological and Operative Severity Score for the Enumeration of Mortality and Morbidity (POSSUM) in patients over the age of 70 was classified as no predictive value (p = 0.217 and p = 0.063, respectively). Conclusion: The mE-PASS and POSSUM models are ineffective in predicting postoperative morbidity in patients above the age of 70. In biliary tract cancer (BTC) patients undergoing radical operation, a combination of E-PASS and perioperative parameters generates a reasonable prediction value for severe complications.

A broad cuproptosis landscape in inflammatory bowel disease.

Background: Cuproptosis, a genetic process of copper-dependent cell death linked to mitochondria respiration, demonstrates its correlation with inhibiting tumoral angiogenesis and motility. Recent studies have developed systematic bioinformatics frameworks to identify the association of cuproptosis with tumors but any non-neoplastic diseases. Therefore, against the background of an increased incidence of inflammatory bowel disease (IBD), the landscape of cuproptosis regulation in IBD is a critical need to be investigated. Methods: The differentially expressed cuproptosis-related genes (DECRGs) were identified with human sequencing profiles for four inflammatory digestive disorders. Another four independent IBD datasets from GEO were used as a validation cohort. And experimental mice model provides another validation method. Using single sample gene set enrichment analysis (ssGSEA), receiver operating characteristic (ROC) curve, CIBERSORT, and consensus clustering algorithms, we explored the association between immune score and cuproptosis-related genes, as well as the diagnostic value of these genes. Molecular docking screened potential interaction of IBD drugs with the structural regulator by Autodock Vina. Results: Cuproptosis-related regulators exhibited extensive differential expression in Crohn's Disease (CD), Ulcerative Colitis (UC), Celiac Disease (CEL), and IBD-induced cancer (IBD-CA) that share common differential genes (PDHA1, DBT, DLAT, LIAS). The differential expression of DECRGs was reverified in the validated cohort and immunohistochemistry assay. Moreover, the cell signaling pathways and ontology mainly focused on the mitochondrial respiratory function, which was highly enriched in Gene set enrichment analysis (GSEA). According to ssGSEA and ROC, when considering the four regulators, which showed robust association with immune infiltration in IBD, the area under the ROC (AUC) was 0.743. In addition, two clusters of consensus clustering based on the four regulators exhibit different immune phenotypes. According to molecular docking results, methotrexate gained the highest binding affinity to the main chain of key cuproptosis-related regulators compared with the remaining ten drugs. Conclusion: Cuproptosis-related regulators were widely linked to risk variants, immune cells, immune function, and drug efficacy in IBD. Regulation of cuproptosis may deeply influence the occurrence and development of patients with IBD.

Effects of Plant Growth-Promoting Rhizobacteria on the Growth and Soil Microbial Community of Carya illinoinensis.

Plant growth-promoting rhizobacteria (PGPR) are important members of soil microbial communities. In this study, the effects of several PGPR on the growth of Carya illinoinensis plants, the microbial community composition and soil nutrients were investigated by inoculation tests to identify excellent PGPR strains. The experiment showed that after PGPR application, the plant height, ground diameter, and dry weight of C. illinoinensis were significantly increased compared with those of the control group, and Bacillus velezensis YH20 had the most significant effect in promoting growth (p < 0.05). In addition, all the PGPRs used for inoculation promoted plant root growth, and the Brevibacillus reuszeri MPT17 strain had the most significant promoting effect on plant root growth (p < 0.05). The application of PGPRs also affected the nutrient levels in plants and plant rhizosphere soil. For example, compared with the control, the levels of available phosphorus and potassium in rhizosphere soil and the total potassium content in plant roots were significantly increased under Br. reuszeri MPT17 treatment (p < 0.05). The experiment showed that the relative abundance of Mortierella, Dictyophora, and Bacillus in the rhizosphere soil increased significantly after the application of PGPR (p < 0.05). These genera could effectively improve the rate of soil nutrient use, antagonize plant pathogenic bacteria, and promote plant growth. This study provides basic reference data regarding the use of PGPR to improve the microecological environment and promote the growth and development of C. illinoinensis plants.

The immuneoreaction and antioxidant status of Chinese mitten crab (Eriocheir sinensis) involve protein metabolism and the response of mTOR signaling pathway to dietary methionine levels.

To study the effects of dietary methionine on growth performance, immunity, antioxidant capacity, protein metabolism, inflammatory response and apoptosis factors in Chinese mitten crabs (Eriocheir sinensis). Five diets with different methionine levels (0.63%, 0.85%, 1.06%, 1.25% and 1.47%) were fed to E. sinensis for 8 weeks. Results showed that in the 1.25% Met group, both growth performance and feed utilization were significantly increased. The crude protein content of crab muscle in the 1.06% and 1.25% Met groups was significantly higher than that in the control group. The immune and antioxidant enzyme activities, as well as gene expression levels of anti-lipopolysaccharide factor 1 (ALF1), Crustin-1, prophenoloxidase (proPO), cap 'n' collar isoform C (CncC) in 1.25% Met group were significantly higher than other groups. The activities of adenosine deaminase (ADA) and glutamate transaminase (GPT) in serum decreased first and then increased with the increase of methionine content, while the changes of ADA and GPT in hepatopancreas increased first and then decreased. 1.25% Met group exhibited significantly increased levels of GOT, GPT, and ADA compared to the control group. 1.25% Met diet group significantly up-regulated protein synthesis and anti-apoptotic factors, and significantly down-regulated inflammatory and pro-apoptotic factors in hepatopancreas. At 1.25% in the diet, methionine was found to boost E. sinensis growth, muscle protein deposition and immunity, as well as its antioxidant capacity. Combined with the above results, based on the expression of factors involved in the mammalian target of rapamycin (mTOR) signaling pathway and the mitogen-activated protein kinase (MAPK) signaling pathway, it is proved that methionine can not only promote protein metabolism, improve feed utilization, but also alleviate the inflammatory response and apoptosis caused by oxidative stress in the body.

Evaluation of leachate production and level in municipal solid waste landfills considering secondary compression.

Landfilled municipal solid waste (MSW) in developing countries generally produces a large amount of leachate due to high moisture content. The estimation of leachate production and level is of great importance to the capacity design of leachate treatment plants and the stability analysis of landfills. In this study, the leachate production ratios (the ratio of leachate mass to waste mass) in different countries and cities were first summarized to reveal the basic status of leachate generation. Then, a model was established to calculate the leachate production and level that considers the rainfall infiltration and the water released from MSW due to both primary and secondary compression (ignored in previous models). Finally, the proposed model was used in a case study of Laogang Landfill in Shanghai, China. It was found that the leachate proportion produced by compression was much higher compared with that produced by rainfall infiltration, ranging from 49 (rainy season) to 93% (dry season). The leachate released from waste due to secondary compression accounted for a high proportion (up to 25%) of the total leachate production, especially for aged MSW. The calculated leachate discharge amount and leachate level were close to the measured values because the possible low permeability layer at the bottom of the landfill was considered in this model.

Exosomes derived from inflammatory myoblasts promote M1 polarization and break the balance of myoblast proliferation/differentiation.

BACKGROUND: Acute muscle injuries are one of the most common injuries in sports. Severely injured muscles are prone to re-injury due to fibrotic scar formation caused by prolonged inflammation. How to regulate inflammation and suppress fibrosis is the focus of promoting muscle healing. Recent studies have found that myoblasts and macrophages play important roles in the inflammatory phase following muscle injury; however, the crosstalk between these two types of cells in the inflammatory environment, particularly the exosome-related mechanisms, had not been well studied. AIM: To evaluate the effects of exosomes from inflammatory C2C12 myoblasts (IF-C2C12-Exos) on macrophage polarization and myoblast proliferation/differentiation. METHODS: A model of inflammation was established in vitro by lipopolysaccharide stimulation of myoblasts. C2C12-Exos were isolated and purified from the supernatant of myoblasts by gradient centrifugation. Multiple methods were used to identify the exosomes. Gradient concentrations of IF-C2C12-Exos were added to normal macrophages and myoblasts. PKH67 fluorescence tracing was used to identify the interaction between exosomes and cells. Microscopic morphology, Giemsa stain, and immunofluorescence were carried out for histological analysis. Additionally, ELISA assays, flow cytometry, and western blot were conducted to analyze molecular changes. Moreover, myogenic proliferation was assessed by the BrdU test, scratch assay, and CCK-8 assay. RESULTS: We found that the PKH-67-marked C2C12-Exos can be endocytosed by both macrophages and myoblasts. IF-C2C12-Exos induced M1 macrophage polarization and suppressed the M2 phenotype in vitro. In addition, these exosomes also stimulated the inflammatory reactions of macrophages. Furthermore, we demonstrated that IF-C2C12-Exos disrupted the balance of myoblast proliferation/differentiation, leading to enhanced proliferation and suppressed fibrogenic/myogenic differentiation. CONCLUSION: IF-C2C12-Exos can induce M1 polarization, resulting in a sustained and aggravated inflammatory environment that impairs myoblast differentiation, and leads to enhanced myogenic proliferation. These results demonstrate why prolonged inflammation occurs after acute muscle injury and provide a new target for the regulation of muscle regeneration.

Exome-based preconception carrier testing for consanguineous couples in China.

OBJECTIVE: To evaluate the utility of clinical exome sequencing (ES)-based carrier screening in Chinese consanguineous couples. METHODS: Consanguineous couples were screened for autosomal recessive (AR) disorders using the clinical ES of 5000 genes associated with human diseases. RESULTS: We recruited 14 couples who elected to have sequencing. One couple was related as first cousins and 13 as second cousins. Both partners carrying the same pathogenic variant were detected in four couples. One couple was found in which one partner carried a splice variant, and the other had a missence variant of the same gene. These five couples were identified as being at risk of having a child affected by an AR disorder. CONCLUSION: Our study demonstrates that ES-based preconception screening yields a clinical value for Chinese consanguineous couples. It enables to detect at-risk couples for rare AR diseases.

Axillary Paget disease with a visible satellite: a case report and literature review.

BACKGROUND: Extramammary Paget disease (EMPD) is an uncommon malignancy affecting apocrine gland-bearing skin, such as vulvar, perianal, axillary and penoscrotal areas. Paget cells are sometimes detected outside clinical border in a phenomenon called subclinical extension. Satellite is one of the patterns of subclinical extension which is likely to be invisible. The standard management strategy for EMPD without distant metastasis is a complete surgical removal, sometimes called wide local excision. However, there is no consensus regarding surgical margin width to decrease the high recurrence rate. Here we describe the first macroscopically visible satellite of axillary EMPD and wide local excision of both main lesion and the satellite lesion with only 0.5 cm margin, succeeded by a short review of the literature. CASE PRESENTATION: A 48-year-old female with a red macule in the right axilla was presented to our clinic. A well-demarcated 4 cm × 3 cm erythematous plaque was observed in the right axilla, and a similar lesion measuring 0.5 cm × 0.3 cm was found 3.5 cm away from the primary site. Breast and axillary node examination was unremarkable. Biopsy of the large plaque revealed Paget disease, then we performed a local extended excision of both lesions with a 0.5 cm margin, all margins negative indicated, by frozen pathology. Pathology revealed the nature of the satellite beside the main lesion also as Paget disease. The patient is currently followed-up for 20 months and has shown no signs of recurrence, with normal shoulder motor function. CONCLUSION: We have report the first visible satellite of extramammary Paget disease, indicating the necessity of an extended local resection of both the main leision and the satellite lesion. Considering the anatomical structure of axillary Paget disease, a 0.5 cm negative surgical margin indicated by frozen pathology might be sufficient to sustain the shoulder motor function.