Regulation of Hepatocytes in G0 and G1 Phases by NOTCH3 mRNA, miR-369-3p, and rno-Rmdn2_0006 during the Initial Stage of Rat Liver Regeneration.

The key event of liver regeneration initiation (LRI) is the switch of hepatocytes from the G0 phase to the G1 phase. This study aimed to use the data from large-scale quantitatively detecting and analyzing (LQDA) to reveal the regulation of hepatocytes in the G0 or G1 phase by competing endogenous RNAs (ceRNAs) during LRI. The hepatocytes of the rat liver right lobe were isolated 0, 6, and 24 h after partial hepatectomy. Their ceRNA expression level was measured using LQDA, and the correlation among their expression, interaction, and role was revealed by ceRNA comprehensive analysis. The expression of neurogenic loci notch homologous protein 3 (NOTCH3) mRNA was upregulated in 0 h, but the expression of miR-369-3p and rno-Rmdn2_0006 of hepatocytes did not change significantly. Meanwhile, the expression of the G0 phase-related gene CDKN1c was promoted by NOTCH3 upregulation, and the expression of the G1 phase-related gene PSEN2 was inhibited by NOTCH3 downregulation. On the contrary, the expression of NOTCH3 mRNA and rno-Rmdn2_0006 was upregulated at 6 h, but the expression of miR-136-3p was downregulated. The expression of the G1 phase-related genes CHUK, DDX24, HES1, NET1, and STAT3 was promoted by NOTCH3 upregulation, and the expression of the G0 phase-related gene CDKN1a was inhibited by NOTCH3 downregulation. These results suggested that the ceRNAs and the NOTCH3-regulated G0 phase- and G1 phase-related genes showed a correlation in expression, interaction, and role. They together regulated the hepatocytes in the G0 phase at 0 h and in the G1 phase at 6 h. These findings might help understand the mechanism by which ceRNA together regulated the hepatocytes in the G0 or G1 phase.

3-Hydroxybutyrate helps crayfish resistant to Vibrio parahaemolyticus infection in versatile ways.

The bacterial storage compound poly-ß-hydroxybutyrate (PHB) is a potential bio-control agent in aquaculture. It has been reported that PHB benefit to the survival and growth, and improve their immunity of aquatic animals. However, the cellular and molecular regulation mechanisms of PHB in immunity process remain unclear. This study investigated the immune mechanism of hemocytes regulated by Halomonas-PHB (PHB-HM) and PHB monomer 3-HB. Red claw crayfish Cherax quadricarinatus was used as the experimental animal in cytological study. Fluorescence microscopy and flow cytometry (FCM) analysis indicated that PHB-HM labeled with fluorescein isothiocyanate (FITC) could be engulfed by granulocytes (Gs) and semi-granulocytes (SGs) upon in vitro incubation. Transmission electron microscopy (TEM) further showed the ongoing degradation of PHB granules inside Gs and SGs after the injection of PHB-HM into crayfish sinus, but phagocytosis of PHB-HM by hyalinocyte (H) was not observed. Therefore, Gs and SGs are considered the main effector cells of cellular immunity induced by PHB-HM, and SGs likely played a particular important role in this process. To study the biosafety and molecular mechanism of PHB monomer 3-HB, hemocyte viability and expression of the related genes were determined after being exposed to 0-1 mg/mL of 3-HB, and Vibrio parahaemolyticus (VP) was used as the pathogenic bacterium. The results confirmed that 3-HB had no toxic effect on hemocytes by means of cell viability assay, and supplementation with 1 mg/mL of 3-HB suppressed the growth rate of VP in TSB medium. Moreover, injection of 3-HB into the blood sinus of crayfish remarkably improved the phagocytic rate of Gs and SGs on VP. Furthermore, transcriptome assay was designed to illuminate the molecular mechanism of 3-HB regulation using red swamp crayfish Procambarus clarkii as experimental animals. RNA-seq analysis and qRT-PCR verification revealed that the microtubule and cytoskeleton-related genes were high expressed 3 h after 3-HB injection, indicating both genes might involve in building up the innate immunity. In summary, bacterial storage PHB could be phagocytosed by main effector blood cells and likely to be degraded within the cells. 3-HB helped the crayfish resistant to pathogens through improving phagocytosis, suppressing the growth of pathogenic bacteria, and increasing the expression of microtubule-related genes. The findings in this work provide cytological and molecular evidence which will facilitate the application of PHB and 3-HB as immune-control agents in farming of aquatic animals.

Clinical characteristics and prognosis of primary hepatolithiasis in hospitalized children.

The purpose of this study is to evaluate the clinical characteristics and prognosis of primary hepatolithiasis in hospitalized children. This retrospective cohort study included 106 hospitalized patients with primary hepatolithiasis at Beijing Children's Hospital. Clinical data were derived from electronic inpatient (2010-2021) and outpatient (2016-2021) medical records. The prognosis was evaluated by outpatient ultrasounds after discharge and telephone interviews performed in December 2022. Intrahepatic bile duct stones in patients enrolled in the study were all found incidentally by abdominal ultrasound during hospitalization, with an incidence of 1.7 per 10,000 hospitalized children. The mean age at diagnosis was 9.3 ± 3.6 years, with male predominance (69/106, 65.1%). The right lobe of the liver (80/106, 75.5%) was preferentially affected. All patients underwent conservative observation. Seventy-eight patients (78/106, 73.6%) were followed up with a mean follow-up age of 17.1 ± 5.0 years, and 4 (4/78, 5.1%) had intermittent abdominal pain. From 2016 to 2021, 32 patients were diagnosed with primary hepatolithiasis, and follow-up abdominal ultrasounds were performed in 20 of them (20/32, 60.0%) with a median time of 2.1 (0.1, 3.5) years. The stones were present in 17 patients (17/20, 85.0%). CONCLUSION: Primary hepatolithiasis in hospitalized children is rare, almost found accidentally; mostly affects the right lobe of the liver; and can be conservatively observed without surgical treatment in childhood. WHAT IS KNOWN: • Primary hepatolithiasis in adults often presents with severe clinical symptoms and requires hepatectomy.. • There are few studies on primary hepatolithiasis in children. WHAT IS NEW: • Primary hepatolithiasis in children is mostly found accidentally by abdominal ultrasound without associated symptoms during hospitalization. • Children with accidental primary hepatolithiasis can be conservatively observed without surgical treatment in childhood.

Analysis of Factors Influencing the Prognosis of Periodontal Disease Treated with Host Modulation and the Development of a Predictive Model.

Objective: To explore the effect of host regulation (HMT) therapy on periodontal disease and construct prognosis prediction model. Methods: The clinical data of 106 patients who received HMT treatment in our hospital from January 2020 to January 2023 were retrospectively analyzed. The changes of plaque index (PLI), periodontal pocket depth (PD), tooth looseness (TM), attachment loss (AL) and gingival bleeding index (BI) were observed before and after treatment. According to the clinical efficacy of patients after treatment, they were divided into effective group and ineffective group. Multivariate Logistic regression model was used to analyze the relevant factors affecting the prognosis of patients., and a predictive model of the factors affecting the prognosis of periodontal disease was constructed to predict the diagnostic value of the prognostic outcome of periodontal disease using the relative operating characteristic (ROC) curve. Results: After HMT treatment, PLI, PD, AL and BI were significantly reduced in all patients, while TM(grade II and III) patients were decreased (P < .05). After 4 weeks of treatment, a total of 74 patients were included in the effective group. Univariate and multivariate analysis showed that oral hygiene, early use of broad-spectrum antibiotics, periodontal bone loss shape, gingival flap thickness and alveolar bone resorption type were independent factors influencing the prognosis of HMT (P < .001). ROC results showed that the area under the curve (AUC), sensitivity and specificity of the above independent influencing factors combined to predict the clinical efficacy of HMT in patients with periodontal disease were higher. Conclusion: HMT has a good clinical effect in the treatment of periodontal disease, but there are still cases of poor prognostic effect. The combination of independent factors affecting the prognosis of HMT has a good predictive value for the clinical efficacy of patients with periodontal disease after HMT treatment.

Source-based health risk assessment of heavy metal contamination in soil: a case study from a polymetallic mining region in Southeastern Hubei, Central China.

To conduct a precise health risk assessment of heavy metals (HMs) in soil, it is imperative to ascertain the primary sources of potential health risks. In this study, we conducted comprehensive measurements of HMs, specifically focusing on the accumulation of Cu, Cd, Sb, Zn, and Pb in local soil, which may pose threats to environmental quality. To achieve our objective, we employed a method that combines positive matrix factorization with a health risk assessment model to quantify the health risks associated with specific sources. The results obtained from the geo-accumulation index indicate that the majority of HMs found in the local soil are influenced by anthropogenic activities. Among these sources, local industrial-related activities contributed the largest proportion of HMs to the soil at 34.7%, followed by natural sources at 28.7%, mining and metallurgy-related activities at 28.2%, and traffic-related activities at 8.40%. Although the non-carcinogenic and carcinogenic risks associated with individual HMs were found to be below safety thresholds, the cumulative health risks stemming from total HMs exceeded safety limits for children. Moreover, the unacceptable health risks for children originating from industrial-related activities, natural sources, and mining and metallurgy-related activities were primarily concentrated in proximity to mining sites and industrial areas within the local region. This investigation furnishes valuable insights that can aid governmental authorities in formulating precise control policies to mitigate health threats posed by soils in polymetallic mining areas.

Microbial community structure analyses and cultivable denitrifier isolation of Myriophyllum aquaticum constructed wetland under low C/N ratio.

With the rapid expansion of livestock production, the amount of livestock wastewater accumulated rapidly. Lack of biodegradable organic matter makes denitrification of livestock wastewater after anaerobic digestion more difficult. In this study, Myriophyllum aquaticum constructed wetlands (CWs) with efficient nitrogen removal performance were established under different carbon/nitrogen (C/N) ratios. Analysis of community composition reveals the change of M. aquaticum CWs in microbial community structure with C/N ratios. The proportion of Proteobacteria which is one of the dominant phyla among denitrifier communities increased significantly under low C/N ratio conditions. Besides, to obtain cultivable denitrifier that could be added into CWs in situ, 33 strains belonging to phylum Proteobacteria were isolated from efficient M. aquaticum CWs, while the best-performing denitrification strain M3-1 was identified as Bacillus velezensis JT3-1 (GenBank No. CP032506.1). Redundancy analysis and quadratic models showed that C/N ratio had significant effects on disposal of nitrate (NO3--N) and the strains isolated could perform well in denitrification when C/N ratio is relatively low. In addition, they have relatively wide ranges of carbon sources, temperature and a high NO3- removal rate of 9.12 mg/(L·hr) at elevated concentrations of 800 mg/L nitrate. Thus, strains isolated from M. aquaticum CWs with low C/N ratio have a practical application value in the treatment of nitrate-containing wastewater. These denitrifying bacteria could be added to CWs to enhance nitrogen removal efficiency of CWs for livestock wastewater with low C/N ratio in the future.

Comparison of the EAONO/JOS, STAMCO and ChOLE cholesteatoma staging systems in the prognostic evaluation of acquired middle ear cholesteatoma in children.

OBJECTIVE: To compare the performance of the EAONO/JOS, STAMCO, and ChOLE Cholesteatoma Staging Systems in prognostic evaluation of children acquired middle ear cholesteatoma after primary surgery and identify the other factors that could predict cholesteatoma recidivism. And the correlation between the staging and the recidivism of cholesteatoma after grouping according to operation was evaluated. METHODS: A total of 123 ears of 118 patients that underwent surgery for primary cholesteatoma from November 2008 to May 2020 were included in this retrospective study, and then classified and staged according to the EAONO/JOS, STAMCO, and ChOLE cholesteatoma staging system, respectively. Each indicator involved in the system above was analyzed separately to evaluate its prognostic value for cholesteatoma recidivism. RESULTS: The type of surgical procedure performed (P = 0.020) was shown to be associated with cholesteatoma recidivism. Cholesteatoma location the supratubal recess (S1) (P = 0.026, HR = 3.614, 95% CI 1.137, 7.945), and the sinus tympani (S2) (P = 0.004, HR = 4.208, 95% CI 1.574, 11.250) were shown to be significantly associated with disease recidivism. When focusing on the CWU operation group, ossicular chain status in STAMCO stage (P = 0.043) and in the ChOLE stage (P = 0.018) were significantly associated with cholesteatoma recidivism. The results had shown no association between the three stages and cholesteatoma recidivism in the CWD and endoscopic surgery groups. CONCLUSIONS: Based on our study, the EAONO/JOS, STAMCO, and ChOLE Classifications have limited value in predicting cholesteatoma recidivism, in acquired middle ear cholesteatoma in children. Adding the pathological status of the ossicular chain may be useful for predicting the recidivism of cholesteatoma. Additional validation studies are entailed to definitively assess the clinical utility of these classifications.

Overview of MicroRNAs as Diagnostic and Prognostic Biomarkers for High-Incidence Cancers in 2021.

MicroRNAs (miRNAs) are small non-coding RNAs (ncRNAs) about 22 nucleotides in size, which play an important role in gene regulation and are involved in almost all major cellular physiological processes. In recent years, the abnormal expression of miRNAs has been shown to be associated with human diseases including cancer. In the past ten years, the link between miRNAs and various cancers has been extensively studied, and the abnormal expression of miRNAs has been reported in various malignant tumors, such as lung cancer, gastric cancer, colorectal cancer, liver cancer, breast cancer, and prostate cancer. Due to the high malignancy grade of these cancers, it is more necessary to develop the related diagnostic and prognostic methods. According to the study of miRNAs, many potential cancer biomarkers have been proposed for the diagnosis and prognosis of diseases, especially cancer, thus providing a new theoretical basis and perspective for cancer screening. The use of miRNAs as biomarkers for diagnosis or prognosis of cancer has the advantages of being less invasive to patients, with better accuracy and lower price. In view of the important clinical significance of miRNAs in human cancer research, this article reviewed the research status of miRNAs in the above-mentioned cancers in 2021, especially in terms of diagnosis and prognosis, and provided some new perspectives and theoretical basis for the diagnosis and treatment of cancers.

Clinical features and outcomes of bowel perforation in primary pediatric gastrointestinal lymphoma.

BACKGROUND: Whether surgery can improve the prognosis of patients with primary pediatric gastrointestinal lymphoma (PPGL) who experienced bowel perforation remains controversial. This study aimed to evaluate the prognosis of such patients. METHODS: Nine patients pathologically diagnosed with PPGL who experienced perforation at our center between January 2010 and December 2020 were enrolled and divided into two groups: those with perforation during (n = 4) and before (n = 5) chemotherapy. Their medical records were reviewed, and long-term follow-up was conducted by telephone in February 2021. RESULTS: All patients with perforation during chemotherapy were diagnosed with PPGL in the outpatient department. The mean time from outpatient visit to chemotherapy was 17.3 ± 6.1 days. Two patients experienced perforation during the first chemotherapy regimen and received conservative treatment, while the others developed perforation after multiple chemotherapy regimens and underwent surgery. All of the patients received regular chemotherapy and survived for a mean follow-up time of 3.8 ± 1.9 years. No patient with perforation before chemotherapy had a definite diagnosis in the outpatient department. Among these patients, 4 experienced perforation and underwent surgery, of whom 3 developed perforation-related complications and died; the other recurred after chemotherapy. Only the patient who received conservative treatment was diagnosed with PPGL before chemotherapy, received regular chemotherapy, and survived without a recurrence for 1.0 year. CONCLUSION: Prompt diagnosis and chemotherapy improve the prognosis of PPGL. Surgery does not affect the prognosis of patients with perforation during chemotherapy but may accelerate disease progression in patients with perforation before chemotherapy.

circRNA-14723 promotes hepatocytes proliferation in rat liver regeneration by sponging rno-miR-16-5p.

Circular RNA (circRNA) is a subclass of noncoding RNA (ncRNA) detected within mammalian tissues and cells. However, its regulatory role during the proliferation phase of rat liver regeneration (LR) remains unreported. This study was designed to explore their regulatory mechanisms in cell proliferation of LR. The circRNA expression profile was detected by high-throughput sequencing. It was indicated that 260 circRNAs were differentially expressed during the proliferation phase of rat LR. Among them, circ-14723 displayed a significantly differential expression. We further explored its regulatory mechanism in rat hepatocytes (BRL-3A cells). First, EdU, flow cytometry and western blot (WB) indicated that knocking down circ-14723 inhibited BRL-3A cells proliferation. Second, RNA-Pulldown and dual-luciferase report assay showed that circ-14723 could sponge rno-miR-16-5p. At last, WB showed that the reported target genes of rno-miR-16-5p, CCND1, and CCNE1 were downregulated after knocking down circ-14723. In conclusion, we found that circ-14723 exerted a critical role in G1/S arrest to promote cell proliferation via rno-miR-16-5p/CCND1 and CCNE1 axis in rat LR. This finding further revealed the regulatory mechanisms of circRNA on cell proliferation of LR, and might provide a potential target for clinical problems.

Serine peptidase inhibitor Kazal type III (SPINK3) promotes BRL-3A cell proliferation by targeting the PI3K-AKT signaling pathway.

The serine protease inhibitor, Kazal type III (SPINK3), is a trypsin inhibitor associated with liver disease, which highly overexpresses in a variety of cancers. In one of our previous studies of our laboratory, Spink3 was observed to be significantly upregulated in rat liver regeneration (LR) via a gene expression profile. For the current study, rat hepatocyte BRL-3A cells were treated by gene addition/interference, and the addition of the exogenous rat recombinant protein SPINK3. It was revealed that both the overexpression of endogenous Spink3 and addition of exogenous rat recombinant SPINK3 (rrSPINK3) significantly promoted the cell proliferation of BRL-3A cells, whereas cell proliferation was inhibited when Spink3 was interfered. Furthermore, quantitative reverse transcription polymerase chain reaction and western blot results revealed that three signaling pathways, including extracellular-signal-regulated kinase 1/2 (ERK1/2), Janus kinase (JAK)-signal transducer and activator of transcription (STAT), and phosphatidylinositol-3-kinase (PI3K)-protein kinase B (AKT), as well as their related genes, were altered following endogenous Spink3 addition/interference. Also, the PI3K-AKT and SRC-p38 pathways and their related genes were modified following exogenous SPINK3 treatment. Among them, the common signaling pathway was PI3K-AKT pathway. We concluded that SPINK3 could activate the PI3K-AKT pathway by enhancing the expression of AKT1 to regulate the proliferation of BRL-3A cells. This study may contribute to shedding light on the potential mechanisms of SPINK3 that regulate the proliferation of BRL-3A cells.

A combined proteomic and metabolomic analyses of the priming phase during rat liver regeneration.

By comparing differentially abundant proteins and metabolites, the protein expression, metabolic changes and metabolic regulation mechanisms during the priming phase of liver regeneration (LR) were investigated. We combined proteomic analysis via isobaric tags for relative and absolute quantification (iTRAQ) with metabolomic analysis via nontargeted liquid chromatography-mass spectrometry (LC-MS). LC-MS was used to examine 29 energy metabolites expression alterations in targeted metabolomics. A total number of 441 differentially expressed proteins and 65 metabolites were identified. PSMB10, PSMB5, RCG_63409, PSME4 and PSMB7 were key node proteins, these proteins are involved in the proteasome pathway. The most strongly enriched transcription factor motif was TP63. These results point out a critical role of the proteasome pathway (defense mechanisms) and of TP63 (metabolic regulator) as the key transcription factor during the priming phase of LR. Metabolomic and metabolite analysis showed that profiling indicates upregulation of arginine biosynthesis and glycolysis as the main ATP-delivering pathway. Integrative proteomic and metabolomic analysis showed that biomolecular changes were primarily related to the neurological disease, cell death and survival and cell morphology. What's more, neurotransmitters may play an important role in the regulation of LR.

Polycyclic aromatic hydrocarbons in railway stations dust of the mega traffic hub city, central China: Human health risk and relationship with black carbon.

Twenty dust samples collected from Wuchang and Wuhan Railway Stations, the biggest transport stations in the mega traffic hub city in Central China, were analyzed for polycyclic aromatic hydrocarbons (PAHs) to investigate the concentration, sources apportionment, and relationship with black carbon (BC) and assess the health risk. The results suggested that the concentrations of PAHs, BC and TOC in Wuhan Railway Station (WHRS) (PAHs = 5940 ± 1920 ng g-1, BC = 53.2 ± 23.1 mg g-1 and TOC = 80.7 ± 44.4) were twice higher than those in Wuchang Railway Station (WCRS) (PAHs = 2580 ± 1630 ng g-1, BC = 20.4 ± 14.3 mg g-1 and TOC = 33.9 ± 20.1 mg g-1). Moreover, the 3 - and 4 - rings PAHs were major PAHs in railway station dust. The composition pattern of PAHs in these railway station dusts had a common characteristic with HMW-PAHs contribution. The results of source identification revealed that different local development features and energy consumption of trains would influence the sources of PAHs and BC. PAHs and BC were most likely related to industrial activities in WHRS. Coal and biomass combustion may influence the PAHs components and BC distribution in WCRS. Moreover, BC had played an important role in retaining PAHs in urban railway stations. Especially in WHRS, BC would more likely to absorb the high molecular weight PAHs, such as 4 -ring (p<0.05), 5 -ring (p<0.05) and 6 -ring (p<0.05) PAHs; while BC just played limited roles in the binding of volatile and semi-volatile organic pollutants, such as 2 -ring and 3 -ring PAHs. With the coexistence of BC and PAHs, passengers would face significant potential health risks by exposure to toxic dust in railway stations, especially for children. The cancer risk in WHRS was almost twice higher than that in WCRS, and it would tend to be stable by a semi-confined structure in the platform area.

Comprehensive analysis of lncRNA-miRNA-mRNA during proliferative phase of rat liver regeneration.

This study aims to reveal the regulatory mechanism of lncRNAs-miRNAs-mRNAs network during the proliferative phase of liver regeneration (LR). High-throughput sequencing technology was performed, and a total of 1,738 differentially expressed lncRNAs (DE lncRNAs), 167 known differentially expressed miRNAs (DE miRNAs), and 2,727 differentially expressed mRNAs were identified. Then, the target DE lncRNAs and DE mRNAs regulated by the same miRNAs were screened and a ceRNA regulatory network containing 32 miRNAs, 107 lncRNAs, and 270 mRNAs was constructed. Insulin signaling pathway, pyrimidine metabolism, axon guidance, carbohydrate digestion and absorption, and pyruvate metabolism were significantly enriched in the network. Through literature review and the regulatory relationship between lncRNAs and miRNAs, nine core lncRNAs were identified, which might play important roles during the proliferative phase of rat LR. This study analyzed lncRNA-miRNA-mRNA regulatory network for the first time during the proliferative phase of rat LR, providing clues for exploring the mechanism of LR and the treatment of liver diseases.

RNA-seq analysis provides insight into molecular adaptations of Andrias davidianus.

The Chinese giant salamander Andrias davidianus is regarded as an ideal model for studying local adaptations, such as longevity, tolerance to starvation, and cutaneous respiration. Transcriptome analysis is useful for studying the large and complex genomes of amphibians. Based on the coding gene set of adult A. davidianus, dozens of A. davidianus-specific genes were identified and three signaling pathway (JAK-STAT, HIF-1, and FoxO) genes were expanded as compared with other amphibians. The results of the pathway analysis of A. davidianus-specific genes indicated that the molecular adaptation of A. davidianus may have required a more rapid evolution of the immune system. Additionally, for the first time, the gene expressions in different parts of the skin tissue were compared. The results of the comparison analysis demonstrated that lateral skin could be more focused on mucus secretion, dorsal skin on immunity and melanogenesis, and abdominal skin on water and salt metabolism. This study provides the first insight into studying longevity and starvation tolerance in A. davidianus, and offers a basis for further investigation of the molecular mechanisms of adaptations in amphibians.

Effects of glycinin and ß-conglycinin on growth performance and intestinal health in juvenile Chinese mitten crabs (Eriocheir sinensis).

This study investigates the effects of two soybean antigens (glycinin and ß-conglycinin) as an antinutritional substance in the diet on the growth, digestive ability, intestinal health and microbiota of juvenile Chinese mitten crabs (Eriocheir sinensis). The isonitrogenous and isolipidic diets contained two soybean antigens at two levels each (70 and 140 g/kg ß-conglycinin, 80 and 160 g/kg glycinin) and a control diet without ß-conglycinin or glycinin supplementation, and were used respectively to feed juvenile E. sinensis for seven weeks. Dietary inclusion of either glycinin or ß-conglycinin significantly reduced crab survival and weight gain. The crabs fed diets containing soybean antigens had higher malondialdehyde concentrations and lower catalase activities in the intestine than those in the control. The activities of trypsin and amylase in the intestine were suppressed by dietary ß-conglycinin and glycinin. Dietary glycinin or ß-conglycinin impaired the immunity and morphological structure of intestine, especially the peritrophic membrane. The mRNA expression of constitutive and inducible immune responsive genes (lipopolysaccharide-induced TNF-α factor and interleukin-2 enhancer-binding factor 2) increased while the mRNA expression of the main genes related to the structural integrity peritrophic membrane (peritrophin-like gene and peritrophic 2) significantly decreased in the groups with soybean antigen addition. Soybean antigen could also change the intestinal microbial community. The abundance of pathogenic bacteria (Ochrobactrum, Burkholderia and Pseudomonas) increased significantly in both soybean antigen groups. Although pathogenic bacteria Vibrio were up-regulated in the glycinin group, the abundance of Dysgonomonas that degraded lignocellulose and ameliorated the gut environment decreased in the glycinin group. This study indicates that existence of soybean antigens (glycinin or ß-conglycinin) could induce gut inflammation, reshape the community of gut microbiota, and cause digestive dysfunction, ultimately leading to impaired growth in crabs.

Individual and combined effects of ammonia-N and sulfide on the immune function and intestinal microbiota of Pacific white shrimp Litopenaeus vannamei.

In this study, we explored the individual and combined effects of ammonia-N and sulfide stress (1 mg/L sulfide and 15 mg/L ammonia-N) on the oxidation resistance, immune response and intestinal health of Litopenaeus vannamei during 72 h exposure. The total antioxidant capacity (T-AOC), malonaldehyde (MDA) and nitric oxide (NO) content, superoxide dismutase (SOD) and catalase activity (CAT), the immune-relative gene (caspase-3, hsp70 and IMD) expression in hepatopancreas and intestine of L.vannamei and the intestinal microbiota were measured. The result showed that MDA and NO contents in hepatopancreas of L. vannamei in all treatment groups increased and remain were at high levels at the end of the stress exposure. The L. vannamei employ antioxidant defense system by increasing the activities of T-AOC, SOD and CAT enzymes in hepatopancereas and intestine to reduce oxidant damage. More severe damages with combined ammonia-N and sulfide stress to antioxidant systems were observed. The gene expression results also demonstrated that antioxidant capacity of L. vannamei was severely impaired and the apoptosis cell was initiated under the ammonia-N and sulfide stress. In addition, the environmental stress also reshaped the intestinal microbial community structure of L. vannamei that a number of original genera decreased, such as Cellvibrio, Vibrio and Rheinheimera; some new genera increased or appeared, such as Photobacterium in all treatment groups, Arcobacter and Fusibacter in sulfide stress group. Therefore, the health of L. vannamei was severely impacted when exposed to the stress of ammonia nitrogen and sulfide and these two factors can have weak synergic effects.

Comprehensive CircRNA expression profile and selection of key CircRNAs during priming phase of rat liver regeneration.

BACKGROUND: Rat liver regeneration (LR) proceeds along a process of highly organized and ordered tissue growth in response to the loss or injury of liver tissue, during which many physiological processes may play important roles. The molecular mechanism of hepatocyte proliferation, energy metabolism and substance metabolism during rat LR had been elucidated. Further, the correlation of circular RNA (circRNA) abundance with proliferation has recently been clarified. However, the regulatory capacity of circRNA in rat LR remains a fascinating topic. RESULTS: To investigate the regulatory mechanism of circRNA during priming phase of rat LR, high-throughput RNA sequencing technology was performed to unbiasedly profile the expression of circRNA during priming phase of rat LR. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) biological pathway analysis was conducted to predict the functions of differentially expressed circRNAs and their host linear transcripts. Co-expression networks of circRNA-miRNA were constructed based on the correlation analysis between the differentially expressed LR-related circRNAs and the condition of their miRNA binding sites. To excavate the key circRNAs in the early phase of rat LR, we comprehensively evaluated and integrated the relationship of expression level between the circRNAs and the linear transcripts as well as the distribution of miRNA binding sites in circRNA sequences. CONCLUSIONS: This paper is the first to employ the comprehensive circRNA expression profile and to investigate circRNA-miRNA interactions during priming phase of rat LR. Two thousand four hundred twelve circRNAs were detected, and 159 circRNAs deriving from 116 host linear transcripts differentially expressed (p < 0.05). Six significantly changed circRNAs during priming phase of rat LR were screened as key circle molecules, and then were validated by qRT-PCR. This study will lay the foundation for revealing the functional roles of circRNAs during rat LR and help solve the remaining clinical problems.

Serine peptidase inhibitor Kazal type I (SPINK1) promotes BRL-3A cell proliferation via p38, ERK, and JNK pathways.

Serine peptidase inhibitor Kazal type I (SPINK1) has the similar spatial structure as epidermal growth factor (EGF); EGF can interact with epidermal growth factor receptor (EGFR) to promote proliferation in different cell types. However, whether SPINK1 can interact with EGFR and further regulate the proliferation of hepatocytes in liver regeneration remains largely unknown. In this study, we investigated the role of SPINK1 in a rat liver hepatocyte line of BRL-3A in vitro. The results showed the upregulation of endogenous Spink1 (gene addition) significantly increased not only the cell viability, cell numbers in S and G2 /M phase, but also upregulated the genes/proteins expression related to cell proliferation and anti-apoptosis in BRL-3A. In contrast, the cell number in G1 phase and the expression of pro-apoptosis-related genes/proteins were significantly decreased. The similar results were observed when the cells were treated with exogenous rat recombinant SPINK1. Immunoblotting suggested SPINK1 can interact with EGFR. By Ingenuity Pathway Analysis software, the SPINK1 signalling pathway was built; the predicted read outs were validated by qRT-PCR and western blot; and the results showed that p38, ERK, and JNK pathways-related genes/proteins were involved in the cell proliferation upon the treatment of endogenous Spink1 and exogenous SPINK1. Collectively, SPINK1 can associate with EGFR to promote the expression of cell proliferation-related and anti-apoptosis-related genes/proteins; inhibit the expression of pro-apoptosis-related genes/proteins via p38, ERK, and JNK pathways; and consequently promote the proliferation of BRL-3A cells. For the first time, we demonstrated that SPINK1 can associate with EGFR to promote the proliferation of BRL-3A cells via p38, ERK, and JNK pathways. This work has direct implications on the underlying mechanism of SPINK1 in regulating hepatocytes proliferation in vivo and liver regeneration after partial hepatectomy.

Probiotic Lactobacillus casei Zhang ameliorates high-fructose-induced impaired glucose tolerance in hyperinsulinemia rats.

AIM: To evaluate the preventive and therapeutic effects of Lactobacilluscasei Zhang on impaired glucose tolerance (IGT) by using fructose-induced hyperinsulinemia rats. METHODS: Rats were fed 25 % fructose solution for hyperinsulinemia with L.casei Zhang for prevention or therapy. Serum levels of insulin, glucagon-like peptide-2 (GLP-2), osteocalcin, malondialdehyde (MDA), total intestinal bile acids and hepatic glycogen contents were determined by assay kits. The major bacteria from feces and liver expression of adiponectin receptor 2 (AdipoR2), liver X receptor-α (LXR-α), peroxisome proliferator-activated receptor gamma (PPAR-γ) and vitamin K epoxide reductase complex subunit 1 mRNA were assessed by RT-PCR. Pancreas injury was evaluated by histological analysis. RESULTS: Lactobacilluscasei Zhang significantly increased numbers of Lactobacillus and Bifidobacterium and decreased Clostridium in the intestine (p < 0.01). Meanwhile, liver glycogen contents were significantly decreased (p < 0.05). In preventive group, accompanied by significantly lower insulin and GLP-2 levels (p < 0.05), L.casei Zhang prevented rats from an increase in oral glucose tolerance area under curve (AUC) which was significant in hyperinsulinemia group (p < 0.05). In therapeutic group, L.casei Zhang administration possessed improved glucose tolerance (p < 0.05), which were associated with increased osteocalcin level (p < 0.01), improved intestinal bile acids secretion (p = 0.060), decreased serum MDA levels (p < 0.05) and upregulation of LXR-α, PPAR-γ and AdipoR2 gene expression, as well as an increase in Bacteroides fragilis (p < 0.05). CONCLUSIONS: Lactobacilluscasei Zhang administration exert both preventive and ameliorative effect on oral glucose tolerance AUC in IGT rats but may be via different mechanisms. L.casei Zhang could prevent rats from increased AUC through GLP-2 lowering, while the ameliorative effect in high-fructose-fed post-adolescent rats may be via B. fragilis enriched vitamin K2-dependent osteocalcin mechanism in which AdipoR2, LXR-α and PPAR-γ signaling were involved.