Distinct Regulation of ASCL1 by the Cell Cycle and Chemotherapy in Small Cell Lung Cancer.

Small cell lung cancer (SCLC) is an aggressive and lethal malignancy. Achaete-scute homolog 1 (ASCL1) is essential for the initiation of SCLC in mice and the development of pulmonary neuroendocrine cells (PNEC), which are the major cells of origin for SCLC. However, the regulatory mechanism of ASCL1 in SCLC remains elusive. Here, we found that ASCL1 expression gradually increases as the tumors grow in a mouse SCLC model, and is regulated by the cell cycle. Mechanistically, CDK2-CyclinA2 complex phosphorylates ASCL1, which results in increased proteasome-mediated ASCL1 protein degradation by E3 ubiquitin ligase HUWE1 during mitosis. TCF3 promotes the multisite phosphorylation of ASCL1 through the CDK2-CyclinA2 complex and the interaction between ASCL1 and TCF3 protects ASCL1 from degradation. The dissociation of TCF3 from ASCL1 during mitosis accelerates the degradation of ASCL1. In addition, chemotherapy drugs greatly reduce the transcription of ASCL1 in SCLC cells. Depletion of ASCL1 sensitizes SCLC cells to chemotherapy drugs. Together, our study demonstrates that ASCL1 is a cell-cycle-regulated protein and provides a theoretical basis for applying cell-cycle-related antitumor drugs in SCLC treatment. Implications:Our study revealed a novel regulatory mechanism of ASCL1 by cell cycle and chemotherapy drugs in SCLC. Treating patients with SCLC with a combination of ASCL1-targeting therapy and chemotherapy drugs could potentially be beneficial.

Associations of Blood Absolute Neutrophil Count and Cytokines With Cognitive Function in Dementia-Free Participants: A Population-Based Cohort Study.

BACKGROUND: The relationships of neutrophils and cytokines with cognitive dysfunction are poorly defined. We aimed to investigate the association of peripheral blood absolute neutrophil count (ANC) with cognitive function in older adults and to further explore the mediating role of serum cytokines in this association. METHODS: This population-based cohort study included 1 666 dementia-free participants (age ≥60 years) derived from baseline examinations (March-September 2018) of the Multimodal Intervention to Delay Dementia and Disability in Rural China (MIND-China); of these, 1 087 participants completed follow-up examinations in October-December 2019. We used a neuropsychological test battery to assess episodic memory, verbal fluency, attention, and executive function at the baseline and follow-up examinations. We used Mindray BC-6800 automated hematology analyzer to measure ANC and Meso Scale Discovery to measure serum interleukin-6 (IL-6) and eotaxin-3. RESULTS: The linear regression analysis of cross-sectional data at baseline (n = 1 666) suggested that increased ANC was significantly associated with a lower episodic memory z score (ß coefficient: -0.149, 95% CI: -0.274 to -0.023) and lower long-delayed free recall z score (-0.216, -0.361 to -0.070). Serum IL-6 and eotaxin-3 could mediate 16.16% to 20.21% and 7.55% to 9.35%, respectively, of these associations. The analysis of longitudinal data (n = 1 087) showed a J-shaped relationship of ANC with decline in episodic memory z score (p for nonlinear = .049), and a U-shaped relationship between ANC and decline in long-delayed free recall z score (p for nonlinear = .043). CONCLUSIONS: Increased neutrophils are associated with poor cognitive performance and accelerated decline in episodic memory, and the cross-sectional association is partly mediated by serum cytokines.

Corrigendum: Case Report: Surgical treatment of a primary giant epithelioid hemangioendothelioma of the spine with total en-bloc spondylectomy.

[This corrects the article DOI: 10.3389/fonc.2023.1109643.].

Resting heart rate, cognitive function, and inflammation in older adults: a population-based study.

BACKGROUND: Emerging evidence has linked elevated resting heart rate (RHR) with poor cognitive function in older adults, but the mechanisms underlying their association are poorly understood. METHODS: This population-based cross-sectional study included 4510 dementia-free participants (age ≥ 65 years; 56.9% females; 38.3% no formal education) in the baseline examination of the Multidomain Interventions to Delay Dementia and Disability in Rural China study. Of these, 1,386 had data on serum proinflammatory cytokines and adhesion molecules. RHR was measured using 12-lead electrocardiograph. We used the Mini-Mental State Examination (MMSE) and a neuropsychological test battery to assess cognitive function. Data were analyzed using the general linear and restricted cubic splines models. RESULTS: People with high RHR were more likely to have cardiometabolic diseases and worse cognitive function (p < 0.05). There was an inverted J-shaped association of RHR with MMSE and attention scores. Having RHR ≥ 80 bpm (vs. 60-69 bpm) was significantly associated with the multivariable-adjusted ß coefficients of - 0.58 [95% confidence interval (CI), - 1.00, - 0.16] for MMSE score and - 0.08 (- 0.15, - 0.01) for attention score. In the serum biomarker subsample, RHR was linearly associated with serum interleukin-6 (IL-6) (ß coefficient = 0.19; 95%CI 0.14, 0.24), IL-8 (0.08; 0.02, 0.13), IL-10 (0.09; 0.04, 0.15), tumor necrosis factor-α (0.06; 0.01, 0.11), monocyte chemotactic protein-1 (0.09; 0.04, 0.15), intercellular adhesion molecule-1 (0.16; 0.11, 0.22), and vascular cell adhesion molecule-1 (0.11; 0.06, 0.16). CONCLUSIONS: There is an inverted J-shaped association of RHR with attention and global cognition. Poor cognitive function and high RHR may be linked through systemic low-grade inflammation and endothelial injury.

Case report: Surgical treatment of a primary giant epithelioid hemangioendothelioma of the spine with total en-bloc spondylectomy.

Background: Epithelioid hemangioendothelioma (EHE) is an extremely uncommon malignant neoplasm that originates from vascular endothelial or pre-endothelial cells. In this report, we present the case of patient who was diagnosed with a primary giant EHE of the spine and underwent treatment with total en-bloc spondylectomy (TES). Case presentation: A 43-year-old male patient with a history of he presented to our hospital with chronic and progressive back pain. Physical examination revealed weakened sensation of acupuncture and touch on the left costal arch, while relatively normal neurological functions were preserved. Radiological examinations identified a giant destructive soft tissue lesion occupying the T8 vertebral region, with moderate destruction of the pedicle and lamina, as well as the 7th left rib. A preoperative biopsy of the 8th vertebra resulted in a diagnosis of epithelioid hemangioendothelioma(EHE). Postoperative immunohistochemical and pathological reports confirmed the presence of EHE in the left ribs and T8 ribs. The patient underwent resection of the 7th left rib and posterior pedicle screw fixation with 8 pairs of screws and a titanium mesh cage. Subsequently, thoracic en bloc spondylectomy was performed on the T8 vertebra. The patient did not receive radiation or chemotherapy following surgery. Over a period of 3 years, the patient remained free of disease and relapse. Conclusion: The use of transarterial embolization with spherical embolic agents (TES) has been demonstrated to be a safe, effective, and reliable treatment option for hepatic epithelioid hemangioendothelioma (EHE). Nevertheless, it is crucial to conduct long-term follow-up of this patient in order to assess their clinical outcome.

Simultaneous addition of CO2-nanobubble water and iron nanoparticles to enhance methane production from anaerobic digestion of corn straw.

In this research, CO2-nanobubble water (CO2-NBW) and iron nanoparticles (Fe0NPs) were added simultaneously to exploit individual advantages to enhance the methanogenesis process from both the stability of anaerobic digestion (AD) system and the activity of anaerobic microorganism aspects. Results showed that the AD performance was enhanced by supplementing with CO2-NBW or Fe0NPs individually, and could be further improved by simultaneous addition of the two additives. The maximum methane yield was achieved in the CO2-NBW + Fe0NPs reactor (141.99 mL/g-VSadded), which increased by 26.16% compared to the control group. Similarly, the activities of the electron transfer system (ETS) and enzyme were improved. The results of microbial community structure revealed that the addition of CO2-NBW and Fe0NPs could improve the abundance of dominant bacteria (Anaerolineaceae, Bacteroidales, and Prolixibacteraceae) and archaea (Methanotrichaceae and Methanospirillaceae). Additionally, the functional metabolic prediction heatmap indicated that metabolic functional genes favorable for AD of corn straw were enhanced.

Cardiovascular health profiles, systemic inflammation, and physical function in older adults: A population-based study.

We examined the association of modifiable cardiovascular health (CVH) metrics with physical function among rural older adults in China and the potential role of inflammatory mechanisms in the association. This study included 3733 stroke- and dementia-free participants (age ≥65 years; 56.9% women) in the baseline survey of a multimodal intervention study in rural China. From March-September 2018, data were collected via face-to-face interviews, clinical assessments, and laboratory tests. The Short Performance Physical Battery (SPPB) test was performed to assess physical function. We defined six modifiable CVH metrics according to the modified American Heart Association's recommendations. Serum interleukin (IL)-6 was measured in a subsample (n = 1156). Data were analyzed with multiple general linear and logistic regression models and structural equation modeling. Poor physical function (SPPB score ≤9) was defined in 1443 participants. Ideal CVH (vs. poor CVH) was associated with multivariable-adjusted odds ratio of 0.60 (95%CI 0.48-0.75) for poor physical function. Ideal CVH was significantly associated with higher scores on balance, chair stand, and walking speed tests (all p < 0.05). Moreover, ideal CVH profile was associated with lower serum IL-6 (multivariable-adjusted ß=-0.04; 95% CI -0.06, -0.01). Mediation analysis revealed that serum IL-6 accounted for 14% of the association of CVH with total SPPB score and 10% of the association with walking speed score (p < 0.05). This study suggests that an ideal CVH profile is associated with better physical function among stroke- and dementia-free older adults, partly via inflammatory mechanisms. The preventive implications of these findings warrant further investigation in cohort studies.

Supplementation of CO2-nanobubble water to enhance the methane production from anaerobic digestion of corn straw.

Nanobubble water (NBW) could improve methane production from anaerobic digestion (AD) of corn straw without secondary contamination. In this study, the effect of carbon dioxide nanobubble water (CO2-NBW) volumes (0%, 25%, 50%, 75%, 100%) on methane production from corn straw was investigated. The results showed that addition of CO2-NBW could improve methane production and promote substrate degradation in AD process. The highest cumulative methane production of 132.16 mL g-1VSadded was obtained in the 100% CO2-NBW added reactor, which was 17% higher than that in the control group. Additionally, the addition of CO2-NBW could mitigate the sharp decrease in pH by acting as a buffer. CO2-NBW could also enhance microorganism activity throughout the AD process. The electron transport system (ETS) activity was increased by 23%, while the ß-glucosidase, dehydrogenase (DHA), and coenzyme F420 activities were increased by 15%, 23%, and 11%, respectively, at optimum addition of CO2-NBW. Meanwhile, addition of CO2-NBW accelerated the production and consumption of reducing sugar and volatile fatty acids (VFAs), promoting the reduction rates of TS (Total solid) and VS (Volatile solid).

Multiplex Assessment of Serum Chemokines CCL2, CCL5, CXCL1, CXCL10, and CXCL13 Following Traumatic Brain Injury.

Chemokines may promote neuroinflammation following traumatic brain injury (TBI), thereby exacerbating secondary injury. This study was designed to investigate the contributions of chemokines (CCL2, CCL5, CXCL1, CXCL10, and CXCL13) to TBI severity and clinical outcome. Peripheral blood was drawn from 92 TBI patients on admission, and 40 controls were recruited. Serum concentrations of CCL2, CCL5, CXCL1, CXCL10, and CXCL13 on admission were measured by ELISA. Preoperative clinical severity was evaluated using the Glasgow Coma Scale (GCS), and clinical outcome at 90 days post-TBI was evaluated using the Glasgow Outcome Scale (GOS). The associations were evaluated by calculating Spearman's correlation coefficients. A binary logistic regression model was used to identify clinicodemographic factors influencing outcome, and ROC curves were constructed. Serum concentrations of CCL2, CCL5, CXCL1, CXCL10, and CXCL13 were elevated significantly after TBI and negatively correlated with GCS and GOS scores except CCL5. CCL2 may be considered as an independent predictor to predict severity and outcome. Moreover, combination of GCS score, CCL2, and CXCL10 can be a better assessment prognosis of moderate and severe TBI.

KLF9 and EPYC acting as feature genes for osteoarthritis and their association with immune infiltration.

BACKGROUND: Osteoarthritis, a common degenerative disease of articular cartilage, is characterized by degeneration of articular cartilage, changes in subchondral bone structure, and formation of osteophytes, with main clinical manifestations including increasingly serious swelling, pain, stiffness, deformity, and mobility deficits of the knee joints. With the advent of the big data era, the processing of mass data has evolved into a hot topic and gained a solid foundation from the steadily developed and improved machine learning algorithms. Aiming to provide a reference for the diagnosis and treatment of osteoarthritis, this paper using machine learning identifies the key feature genes of osteoarthritis and explores its relationship with immune infiltration, thereby revealing its pathogenesis at the molecular level. METHODS: From the GEO database, GSE55235 and GSE55457 data were derived as training sets and GSE98918 data as a validation set. Differential gene expressions of the training sets were analyzed, and the LASSO regression model and support vector machine model were established by applying machine learning algorithms. Moreover, their intersection genes were regarded as feature genes, the receiver operator characteristic (ROC) curve was drawn, and the results were verified using the validation set. In addition, the expression spectrum of osteoarthritis was analyzed by immunocyte infiltration and the co-expression correlation between feature genes and immunocytes was construed. CONCLUSION: EPYC and KLF9 can be viewed as feature genes for osteoarthritis. The silencing of EPYC and the overexpression of KLF9 are associated with the occurrence of osteoarthritis and immunocyte infiltration.

Dendrobium huoshanense C.Z.Tang et S.J.Cheng: A Review of Its Traditional Uses, Phytochemistry, and Pharmacology.

Dendrobium huoshanense, a traditional medicinal and food homologous plant, belongs to the family Orchidaceae and has a long history of medicinal use. It is reported that the stem of D. huoshanense has a variety of bioactive ingredients such as polysaccharides, flavonoids, sesquiterpenes, phenols, etc. These bioactive ingredients make D. huoshanense remarkable for its pharmacological effects on anti-tumor, immunomodulation, hepatoprotective, antioxidant, and anticataract activities. In recent years, its rich pharmacological activities have attracted extensive attention. However, there is no systematic review focusing on the chemical compositions and pharmacological effects of D. huoshanense. Therefore, the present review aims to summarize current research on the chemical compositions and pharmacological activities of D. huoshanense. This study provides valuable references and promising ideas for further investigations of D. huoshanense.

Nonalcoholic fatty liver disease, serum cytokines, and dementia among rural-dwelling older adults in China: A population-based study.

BACKGROUND AND PURPOSE: Little is known about whether nonalcoholic fatty liver disease (NAFLD) is associated with dementia or the role of serum proinflammatory cytokines in the association. We aimed to investigate the interrelationships of NAFLD, serum cytokines, and dementia among rural-dwelling older adults. METHODS: This population-based cross-sectional study included 5129 participants (aged ≥60 years; 61.79% women) who were living in rural communities and examined in March 2018-September 2018. NAFLD was defined through transabdominal ultrasound examination in the absence of hepatitis B or excessive alcohol consumption. Serum cytokines were measured in a subsample (n = 1686). Dementia, Alzheimer disease (AD), and vascular dementia (VaD) were diagnosed following international criteria. Data were analyzed with logistic regression and mediation models. RESULTS: Of the 5129 participants, 455 (8.87%) were detected with moderate-to-severe NAFLD, and 292 (5.69%) were diagnosed with dementia (188 with AD and 96 with VaD). The multivariable adjusted odds ratios associated with moderate-to-severe (vs. no-to-mild) NAFLD were 2.22 (95% confidence interval [CI] = 1.41-3.49) for all-cause dementia, 1.88 (95% CI = 1.01-3.50) for AD, and 2.62 (95% CI = 1.33-5.17) for VaD. In the cytokine subsample, controlling for multiple potential confounders, moderate-to-severe NAFLD was significantly associated with higher levels of serum monocyte chemotactic protein-1, interleukin-17A, interleukin-6 (IL-6), interleukin-8, and tumor necrosis factor-α (P < 0.05). The mediation analysis showed that IL-6 mediated 12.56% of the association between NAFLD and VaD. CONCLUSIONS: Moderate-to-severe nonalcoholic fatty liver disease is associated with dementia and AD, especially with VaD, among rural-dwelling Chinese older adults, in which the association with VaD is partly mediated by serum inflammatory cytokines.

Screening of genes related to breast cancer prognosis based on the DO-UniBIC method.

BACKGROUND: Early screening is the most effective way to control breast cancer. Due to the lack of accurate biomarkers, early diagnosis of breast cancer is still very difficult. Therefore, it is necessary to discover new candidate genes of breast cancer and improve the early diagnosis and prognosis. METHODS: A DO-UniBIC gene screening method was proposed. First, Disease Ontology (DO) analysis was used to screen out breast cancer related genes from differentially expressed genes, and then the UniBIC algorithm was used to find all gene clusters with the same changing trend based on the longest common subsequence. In addition, an eight-gene prognostic model was constructed to assess the prognostic risk of breast cancer patients. RESULTS: The prognostic analysis of the candidate genomes based on multivariate Cox proportional regression model revealed eight genes that were significantly related to prognosis. The eight genes were ACSL1, CD24, EMP1, JPH3, CAMK4, JUN, S100B and TP53AIP1. Among them, ACSL1 was a new potential breast cancer related gene screened by the DO-UniBIC method. CONCLUSIONS: More comprehensive cancer-related genes can be screened based on the DO-UniBIC method, which can be used as the candidate gene set for prognostic analysis.

Homing peptide combined with DNAzyme-based ELISA-like assay for highly specific and sensitive detection of fibrin.

A highly sensitive and specific ELISA-like chemiluminescence method for detection of fibrin has been developed. In the sensing platform, the homing peptide (CREKA), as recognition molecule, which can specially recognize the fibrin on microtiter plate, combined with G-quadruplex-based DNAzyme to form the probe of G-quadruplex-hemin DNAzyme-CREKA. After the sample solution was coated on the plates, the probe was crosslinked with fibrin through the interaction of CREKA and fibrin. Finally, luminol-H2O2 chemiluminesecence (CL) reaction was exploited for quantitative analysis of fibrin. The liner range for fibrin detection was from 0.112 pmol L-1 to 5.6 pmol L-1 with the detection limit of fibrin as low as 0.04 pmol L-1, based on a signal-to-noise ratio (S/N) of 3. Furthermore, on the basis of the high amplification efficiency of the rolling circle amplification (RCA) reaction, the method enabled to analyze fibrin with a detection limit corresponding to 0.06 fmol L-1, whose sensitivity increased 3 orders of magnitude than that of above method in the absence of RCA reaction. In particular, combined with the separation and washing steps of ELISA, the proposed method possessed higher selectivity, high-throughput and low cost, which shows promise for applications in clinical diagnosis.

Evaluation of immature granulocyte parameters in myeloid neoplasms assayed by Sysmex XN hematology analyzer.

Immature granulocytes (IGs) have significance for the diagnosis of myeloid neoplasms (MNs). The current study aims to use a hematology analyzer to evaluate the accuracy of IG parameters in MNs. Blood specimens from 388 patients with MN, 524 with non-hematological neoplasms (non-HNs), including 109 patients with inflammation and 68 undergoing G-CSF administration, and 500 healthy control subjects were analyzed. IG parameters was assayed by Sysmex XN-9000 (XN) and compared with manual assessments. A high level of agreement between IG% derived from XN and manual measurements for MN patients (r = 0.828, p < 0.0001) was revealed but only a moderate correlation for acute myeloid leukemia patients (AML; r = 0.597; p < 0.0001). Bland-Altman bias analysis was conducted, and the results showed that differences in IG% from XN and manual analysis for MN patients were considered clinically insignificant. ROC analysis demonstrated a good performance of IG# (AUC = 0.842) and IG% (AUC = 0.885) assessed by XN for MN patients with cut-off values of 0.200 × 109/L and 1.95%, respectively. IG parameters from Sysmex XN analyzer are helpful for screening of MNs even though granulocyte morphological abnormalities may interfere with IG parameter accuracy.

Therapeutic effect and immune mechanism of chitosan-gentamicin conjugate on Pacific white shrimp (Litopenaeus vannamei) infected with Vibrio parahaemolyticus.

To explore the disease resistance mechanism of chitosan conjugates, chitosan-gentamicin conjugate (CS-GT) was synthesized and systematically characterized, the immune mechanism of CS-GT on Litopenaeus vannamei infected with Vibrio parahaemolyticus was further explored. The results showed that imine groups in CS-GT were effectively reduced. Dietary supplementation of CS-GT can significantly increase the survival rate, total hemocyte counts, the antioxidant and immune related enzyme activity levels of shrimps (P < 0.05), which are all dose-dependent under the experimental conditions. In addition, CS-GT can protect the hepatopancreas from invading bacteria and alleviate inflammation. Particularly, CS-GT promotes the expressions of legumain (LGMN), lysosomal acid lipase (LIPA) and Niemann-Pick type C2 (NPC2) up-regulated. It is speculated that CS-GT may stimulate the lysosome to phagocytose pathogens more effectively. In conclusions, shrimps fed with CS-GT can produce immune response via lysosome and greatly improve the disease resistance to Vibrio parahaemolyticus.

Shikonin ameliorates lipoteichoic acid­induced acute lung injury via promotion of neutrophil apoptosis.

Shikonin is the major active component in Lithospermum erythrorhizon and has pharmacological effects including reducing inflammation, aiding resistance to bacteria and promoting wound healing. However, the effect of shikonin on lipoteichoic acid (LTA)­induced acute lung injury (ALI) remains to be elucidated. ALI is a serious illness resulting from significant pulmonary inflammation caused by various diseases, such as sepsis, acid aspiration and trauma. The present study found that shikonin significantly attenuated LTA­induced ALI. Following shikonin treatment, the accumulation of pulmonary neutrophils and expression of TNFα, IL­1ß and IL­6 were decreased in mice with LTA­induced ALI. Furthermore, Shikonin promoted neutrophil apoptosis by increasing the activation of caspase­3 and reducing the expression of the antiapoptotic myeloid cell leukemia­1 (Mcl­1) protein. However, shikonin treatment did not influence the expression of B­cell lymphoma­2. The findings of the present study demonstrated that shikonin protected against LTA­induced ALI by promoting caspase-3 and Mcl­1­related neutrophil apoptosis, suggesting that shikonin is a potential agent that can be used in the treatment of sepsis­mediated lung injury.

Giant schwannoma of thoracic vertebra: A case report.

BACKGROUND: It is relatively rare for schwannomas to invade bone, but it is very rare for a large mass to form concurrently in the paravertebral region. Surgical resection is the only effective treatment. Because of the extensive tumor involvement and the many important surrounding structures, the tumor needs to be fully exposed. Most of the tumors are completely removed by posterior combined open-heart surgery to relieve spinal cord compression, restore the stability of the spine and maximize the recovery of nerve and spinal cord function. The main objective of this article is to present a schwannoma that had invaded the T5 and T6 vertebral bodies and formed a large paravertebral mass with simultaneous invasion of the spinal canal and compression of the spinal cord. CASE SUMMARY: A 40-year-old female suffered from intermittent chest and back pain for 8 years. Computed tomography and magnetic resonance imaging scans showed a paravertebral tumor of approximately 86 mm × 109 mm × 116 mm, where the adjacent T5 and T6 vertebral bodies were invaded by the tumor, the right intervertebral foramen was enlarged, and the tumor had invaded the spinal canal to compress the thoracic medulla. The preoperative puncture biopsy diagnosed a benign schwannoma. Complete resection of the tumor was achieved by a two-step operation. In the first step, the thoracic surgeon adopted a lateral approach to separate the thoracic tumor from the lung. In the second step, a spine surgeon performed a posterior midline approach to dissect the tumor from the vertebral junction through removal of the tumor from the posterior side and further resection of the entire T5 and T6 vertebral bodies. The large bone defect was reconstructed with titanium mesh, and the posterior root arch was nail-fixed. Due to the large amount of intraoperative bleeding, we performed tumor angioembolization before surgery to reduce and avoid large intraoperative bleeding. The postoperative diagnosis of benign schwannoma was confirmed by histochemical examination. There was no sign of tumor recurrence or spinal instability during the 2-year follow-up. CONCLUSION: Giant schwannoma is uncommon. In this case, a complete surgical resection of a giant thoracic nerve sheath tumor that invaded part of the vertebral body and compressed the spinal cord was safe and effective.

Enhanced energy recovery via separate hydrogen and methane production from two-stage anaerobic digestion of food waste with nanobubble water supplementation.

This study investigated the enhancement effect of N2- and Air-nanobubble water (NBW) supplementation on two-stage anaerobic digestion (AD) of food waste (FW) for separate production of hydrogen and methane. In the first stage for hydrogen production, the highest cumulative H2 yield (27.31 ± 1.21 mL/g-VSadded) was obtained from FW + Air-NBW, increasing by 38% compared to the control (FW + deionized water (DW)). In the second stage for methane production, the cumulative CH4 yield followed a descending order of FW + Air-NBW (373.63 ± 3.58 mL/g-VSadded) > FW + N2-NBW (347.63 ± 7.05 mL/g-VSadded) > FW + DW (300.93 ± 3.24 mL/g-VSadded, control), increasing by 24% in FW + Air-NBW and 16% in FW + N2-NBW compared to the control, respectively. Further investigations indicate that different gas-NBW may positively impact the different stages of AD process. Addition of N2-NBW only enhanced the hydrolysis/acidification of FW with no significant effect on methanogenesis. By comparison, addition of Air-NBW promoted both hydrolysis/acidification stage and methanogenesis stage, reflecting by the enhanced activities of four extracellular hydrolases at the end of hydrolysis/acidification and coenzyme F420 at the end of methanogenesis, respectively. Results from this work suggest the potential application of Air-NBW in the two-stage AD for efficient renewable energy recovery from FW.

Stress responses of testicular development, inflammatory and apoptotic activities in male zebrafish (Danio rerio) under starvation.

Food deprivation is a severe stress across multiple fields and challenged to organismal development and immune system. Here, adult male zebrafish were used to investigate the starvation stress on organismal development, spermatogenesis, testicular inflammation and apoptosis. Results showed that the biological indexes, blood parameters, and RNA/DNA ratio in testis dramatically decreased after 1-3 weeks of starvation. The testicular architecture was impaired and the spermatogenesis was retarded with increased proportions of spermatogonia and spermatocytes, and decreased proportion of spermatozoa in the starved fish. The mRNA expressions of amh and sycp3 were downregulated, the retinoic acid content increased at later stage of starvation through the transcriptional regulation of aldh1a2 and cyp26a1. Besides, the immune response was elevated with upregulated mRNA and protein expressions of TNF-α, IL-6, and IL-1ß, which indicated the inflammation of opportunistic risk in testis. The apoptotic activity was stimulated, accompanied by differentially upregulated expressions of baxa, casp9, casp3, casp2, and decreased ratio of Bcl-2/Bax in the attenuate testis. Taken together, our findings revealed that the stress responses of testicular development, inflammatory and apoptotic activities in male zebrafish under starvation and pointed out the susceptibility of fish gonad to food fluctuation.