TRIM21 Promotes Oxidative Stress and Ferroptosis through the SQSTM1-NRF2-KEAP1 Axis to Increase the Titers of H5N1 Highly Pathogenic Avian Influenza Virus.

Tripartite motif-containing protein 21 (TRIM21) is involved in signal transduction and antiviral responses through the ubiquitination of protein targets. TRIM21 was reported to be related to the imbalance of host cell homeostasis caused by viral infection. Our studies indicated that H5N1 highly pathogenic avian influenza virus (HPAIV) infection up-regulated TRIM21 expression in A549 cells. Western blot and qPCR results showed that knockdown of TRIM21 alleviated oxidative stress and ferroptosis induced by H5N1 HPAIV and promoted the activation of antioxidant pathways. Co-IP results showed that TRIM21 promoted oxidative stress and ferroptosis by regulating the SQSTM1-NRF2-KEAP1 axis by increasing SQSTM1 K63-linked polyubiquitination under the condition of HPAIV infection. In addition, TRIM21 attenuated the inhibitory effect of antioxidant NAC on HPAIV titers and enhanced the promoting effect of ferroptosis agonist Erastin on HPAIV titers. Our findings provide new insight into the role of TRIM21 in oxidative stress and ferroptosis induced by viral infection.

Plasma LTBP2 as a potential biomarker in differential diagnosis of connective tissue disease-associated interstitial lung disease and idiopathic pulmonary fibrosis: a pilot study.

Few biomarkers distinguish connective tissue disease-associated interstitial lung disease (CTD-ILD) from idiopathic pulmonary fibrosis (IPF). Latent transforming growth factor-ß binding protein-2 (LTBP2), a secreted extracellular matrix protein, is involved in pulmonary fibrosis. However, the role of LTBP2 in differentially diagnosing CTD-ILD and IPF is unclear. In this study, enzyme-linked immunosorbent assays quantified plasma LTBP2 concentrations in 200 individuals (35 healthy controls, 42 CTD patients without ILD, 89 CTD-ILD patients, and 34 IPF patients). CTD-ILD and IPF were further classified based on chest imaging pattern and pulmonary function test results. Plasma LTBP2 levels were significantly elevated in the IPF group compared with the CTD-ILD group. ROC analysis further suggested the possible value of LTBP2 in differentially diagnosing CTD-ILD and IPF. Additionally, CTD-ILD patients with progressive lung fibrosis had higher plasma LTBP2 concentrations than those who did not. Similarly, patients with IPF developing acute exacerbation showed higher plasma LTBP2 levels than those with stable IPF. This is the first study showing that LTBP2 was closely associated with the usual interstitial pneumonia (UIP) pattern in rheumatoid arthritis-associated ILD (RA-ILD). Moreover, the optimal cutoff values of LTBP2 for distinguishing IPF from CTD-UIP/RA-UIP were 33.75 and 38.33 ng/mL with an AUC of 0.682 and 0.681, respectively. Our findings suggest that plasma LTBP2 levels may differentially diagnose CTD-ILD and IPF, and assess their fibrotic activity. Additionally, clinical LTBP2 evaluation may be a great aid to identifying the presence of the UIP pattern in RA-ILD and to discriminating IPF from CTD-UIP, particularly RA-UIP.

The metabolome and bacterial composition of high-moisture Italian ryegrass silage inoculated with lactic acid bacteria during ensiling.

BACKGROUND: With its high nutritional value and productivity, Italian ryegrass as a biomass feedstock constantly supplies rumen degradable nitrogen and digestible fiber to ruminants. However, biofuel production is easily reduced during ensiling due to the high-moisture content of Italian ryegrass, leading to economic losses. Lactic acid bacteria inoculants could improve lignocellulosic degradation and fermentation quality and decrease dry matter loss during the bioprocessing of silage. Therefore, this study analyzed the effects of Lactobacillus buchneri TSy1-3 (HE), Lactobacillus rhamnosus BDy3-10 (HO), and the combination of HE and HO (M) on fermentation quality, bacterial community and metabolome in high-moisture Italian ryegrass silage during ensiling. RESULTS: The results showed that the pH value was significantly lower in the HO groups than in the other treatments at the end of ensiling, and the dry matter and acetic acid contents were significantly higher in the HO group than in the other inoculated groups. All inoculants decreased the diversity of the bacterial community and significantly increased the relative abundance of Lactobacillus. Inoculation with HO significantly improved the concentrations of organic acids, dipeptides, ferulic acid, apigenin, and laricitrin. Compared with Lactobacillus buchneri TSy1-3 (HE), HO significantly upregulated the flavonoid compounds in the flavone and flavonol biosynthesis pathway. CONCLUSIONS: Overall, these findings suggest that inoculation with HO was beneficial for the development of Italian ryegrass as a biomass feedstock, improving fermentation quality, accelerating changes in bacterial community composition and increasing biofunctional metabolites in high-moisture Italian ryegrass silage.

Characterization of mycotoxins and microbial community in whole-plant corn ensiled in different silo types during aerobic exposure.

Silage can be contaminated with mycotoxins and accidental fungi after aerobic exposure. The study assessed the effects of bunker silos (BS), round bales (RB), and silage bags (SB) on the nutritional characteristics, fermentation quality, aerobic stability, mycotoxin levels and microbial communities of whole-plant corn silage (WPCS). After 90 days of fermentation, silages were opened and sampled at 0, 1, 3, 5, 7, and 9 days of exposure. SB group conserved higher lactic acid and dry matter contents and a lower pH value than other groups after 9 days of exposure (p < 0.05). The SB group showed the longest aerobic stability (202 h) among all silages (p < 0.05). The concentrations of aflatoxin B1, trichothecenes and fumonisin B1 were significantly lower in SB after 9 days of exposure (p < 0.05). Acetobacter became the dominant bacteria in BS and RB groups after 5 days of exposure. However, Lactobacillus still dominated the bacterial community in SB group. Acetobacter was positively correlated with pH, acetic acid content, and ammonia-N content (p < 0.05). Lactobacillus was positively correlated with Kazachstania and Candida abundances (p < 0.01) but negatively correlated with Fusarium abundance (p < 0.05). Considering the feed value and food safety of silage in the feeding process, silage bags are recommended for WPCS according to the observed nutritional quality, fermentation index and mycotoxin content.

Phylogenetic and pathogenic characterization of current fowl adenoviruses in China.

In recent years, fowl adenoviruses (FAdVs) continue to outbreak and cause huge economic losses to the poultry industry in China. The homologous recombination accounts for the diversity serotypes of adenovirus. However, the prevalence, recombination and pathogenicity of current FAdVs remain unclear. Herein, the prevalence, phylogenetic feature and pathogenicity of FAdVs in China in 2019 were characterized. Our findings showed that multiple species and serotypes of FAdVs currently circulate in China, including A, C, D and E species, and 1, 2, 4, 8a and 8b serotypes. Notably, the recombination occurred between FAdV-8a and FAdV-8b, and the recombination regions included Hexon, Fiber, ORF19 and ORF20. All five FAdVs replicated effectively in various chicken tissues, and viral shedding peaked at 4-8 dpi. Except CH/GDSZ/1905(FAdV-1/A), the remaining FAdVs caused obvious inclusion body hepatitis (IBH) in 3-week-old specific-pathogen-free (SPF) chickens, of which CH/JSXZ/1905(FAdV-4/C) caused hydropericardium-hepatitis syndrome (HHS) with a mortality rate of 62.5%. Taken together, our findings illustrate the prevalence, recombination and pathogenicity of current FAdVs in China and strengthen surveillance and further pathogenicity studies of FAdVs are extremely urgent.

Effects of different additives on the bacterial community and fermentation mode of whole-plant paper mulberry silage.

The purpose of this study was to investigate the effects of inoculation with two lactic acid bacteria (LAB) strains (Lacticaseibacillus rhamnosus and Lentilactobacillus buchneri) and the addition of four corn flour proportions (0, 3, 6, and 9%) in different treatments, on the composition and function of the bacterial community in whole-plant paper mulberry silage. The different treatments promoted Lactiplantibacillus, Lentilactobacillus, and Lacticaseibacillus growth, but the microbial species responsible for fermentation differed among the treatments. High species diversity and various Gram-negative bacteria, such as Flavitalea sp., Pantoea agglomerans, Acinetobacter pittii, Turicibacter sanguinis, and Ralstonia pickettii, were found in the uninoculated LAB treatments. A beneficial bacterium, Lactobacillus johnsonii, was discovered for the first time in whole-plant paper mulberry silage. LAB inoculation simplifies the microbial community structure, and beneficial Lactobacillus as a key species aggregates in the inoculated treatment group. However, L. rhamnosus inoculation alone may have limited bacteriostatic activity against in whole-plant paper mulberry silage. Compared with silage lacking corn flour, amino sugar and nucleotide sugar metabolism, galactose metabolism, the phosphotransferase system and the pentose phosphate pathway metabolic activity were increased in corn flour-containing silage. Whole-plant paper mulberry can be used as a high-quality silage to provide high-quality feed resources for sustainable ruminant livestock production. Moreover, additive use is necessary for preparing paper mulberry silage.

Phylogenetic and Phylogeographic Analysis of the Highly Pathogenic H5N6 Avian Influenza Virus in China.

The clade 2.3.4.4b H5N8 avian influenza viruses (AIVs) have caused the loss of more than 33 million domestic poultry worldwide since January 2020. Novel H5N6 reassortants with hemagglutinin (HA) from clade 2.3.4.4b H5N8 AIVs are responsible for multiple human infections in China. Therefore, we conducted an epidemiological survey on waterfowl farms in Sichuan and Guangxi provinces and performed a comprehensive spatiotemporal analysis of H5N6 AIVs in China. At the nucleotide level, the H5N6 AIVs isolated in the present study exhibited high homology with the H5N6 AIVs that caused human infections. Demographic history indicates that clade 2.3.4.4b seemingly replaced clade 2.3.4.4h to become China's predominant H5N6 AIV clade. Based on genomic diversity, we classified clade 2.3.4.4b H5N6 AIV into ten genotypes (2.3.4.4bG1-G10), of which the 2.3.4.4bG5 and G10 AIVs can cause human infections. Phylogeographic results suggest that Hong Kong and Jiangxi acted as important epicentres for clades 2.3.4.4b and 2.3.4.4h, respectively. Taken together, our study provides critical insight into the evolution and spread of H5N6 AIVs in China, which indicates that the novel 2.3.4.4b reassortants pose challenges for public health and poultry.

Effect of Different Regions and Ensiling Periods on Fermentation Quality and the Bacterial Community of Whole-Plant Maize Silage.

This study aimed to explore the changes in the microbial community on the silage material surface and during the ensiling process of whole-plant maize in different regions. Whole-plant maize silages were sampled in Ziyun, Guanling, and Weinning counties within warm and humid climate areas in southern China. Silages were sampled at 0, 2, 5, 10, 20, and 45 days during ensiling. The nutritional components, fermentation properties, and microbiomes were examined to evaluate the influence of sampling area and fermentation time on the quality of silage. The results showed that the pH values of all silages significantly decreased (<4.2 at ensiling day 2) during fermentation and all silages achieved satisfactory fermentation at 45 days. Butyric acid was not detected during ensiling, and the contents of acetic acid and ammonia nitrogen in the final silages were below 6 g/kg DM and 50 g/kg total nitrogen, respectively. Weissella was the dominant epiphytic bacteria of raw material in Ziyun and Weinning, while Lactobacillus was prevalent in Guanling. Lactobacillus dominated the ensiling process, and its abundance significantly increased with increasing fermentation time in the three groups. Lactobacillus was negatively correlated with pH of all silages (p < 0.05) and positively correlated with lactic acid, propionic acid and acetic acid (p < 0.05). Furthermore, the bacterial community was significantly correlated with environmental factors. Altitude had a highly positive correlation with the abundance of Stenotrophomonas, Chryseobacterium, and Massilia (p < 0.01), while precipitation was negatively correlated with these bacteria. The humidity and average temperature significantly influenced the Lactobacillus and Weissella abundances of fresh whole-plant maize. During the ensiling process, the silages from three regions had similar bacterial dynamic changes, and the Lactobacillus formed and maintained good fermentation characteristics in whole-plant maize silage.

Cancer Classification with a Cost-Sensitive Naive Bayes Stacking Ensemble.

Ensemble learning combines multiple learners to perform combinatorial learning, which has advantages of good flexibility and higher generalization performance. To achieve higher quality cancer classification, in this study, the fast correlation-based feature selection (FCBF) method was used to preprocess the data to eliminate irrelevant and redundant features. Then, the classification was carried out in the stacking ensemble learner. A library for support vector machine (LIBSVM), K-nearest neighbor (KNN), decision tree C4.5 (C4.5), and random forest (RF) were used as the primary learners of the stacking ensemble. Given the imbalanced characteristics of cancer gene expression data, the embedding cost-sensitive naive Bayes was used as the metalearner of the stacking ensemble, which was represented as CSNB stacking. The proposed CSNB stacking method was applied to nine cancer datasets to further verify the classification performance of the model. Compared with other classification methods, such as single classifier algorithms and ensemble algorithms, the experimental results showed the effectiveness and robustness of the proposed method in processing different types of cancer data. This method may therefore help guide cancer diagnosis and research.

A self-assembled RNA-triple helix hydrogel drug delivery system targeting triple-negative breast cancer.

The major drawbacks of traditional RNA cancer therapies include low cellular uptake in vitro or in vivo, instability of in vivo circulation, nonspecific bio-distribution, and lack of targeting ability, which result in poor silencing efficiency. Herein, we developed a novel RNA-triple-helix hydrogel for the treatment of triple negative breast cancers (TNBCs) by incorporating RNA-triple-helix and siRNA duplexes of CXCR4 into the same RNA nanoparticles with no synthetic polycationic reagents added. The RNA-triple-helix consists of one tumour suppressor miRNA (miRNA-205) and one oncomiR inhibitor (miRNA-221), both of which showed an outstanding effect in synergistically abrogating tumours. The siRNA duplexes of CXCR4 were embedded into the RNA hydrogel to block breast cancer metastasis and conjugation of the LXL-DNA aptamer (apt-DNA-Chol) is an effective target DNA sequence for MDA-MB-231 cells. The self-assembly of the RNA-triple-helix hydrogel exhibited high selectivity of in vitro and in vivo absorption and controlling miRNA expression when compared to free miRNA and RNA transcripts. The well-developed gene delivery system provided a potential treatment with high specificity and selectivity toward TNBCs. This strategy can be implemented in triplex-helix hydrogel design to form novel miRNA combinations to treat various human cancers.

Cancer Classification Based on Support Vector Machine Optimized by Particle Swarm Optimization and Artificial Bee Colony.

Intelligent optimization algorithms have advantages in dealing with complex nonlinear problems accompanied by good flexibility and adaptability. In this paper, the FCBF (Fast Correlation-Based Feature selection) method is used to filter irrelevant and redundant features in order to improve the quality of cancer classification. Then, we perform classification based on SVM (Support Vector Machine) optimized by PSO (Particle Swarm Optimization) combined with ABC (Artificial Bee Colony) approaches, which is represented as PA-SVM. The proposed PA-SVM method is applied to nine cancer datasets, including five datasets of outcome prediction and a protein dataset of ovarian cancer. By comparison with other classification methods, the results demonstrate the effectiveness and the robustness of the proposed PA-SVM method in handling various types of data for cancer classification.

[Analysis of the therapeutic effect of esophagectomy with extended 2-field lymph node dissection for esophageal carcinoma].

OBJECTIVE: To summarize the surgical effect and clinical application value of esophagectomy with extended 2-field lymph node dissection for patients with esophageal carcinoma. METHODS: From June 1987 to December 2008, 1690 patients with esophageal cancer underwent esophagectomy with extended 2-field (thoracic and abdominal) dissection of lymph nodes. Patients with the middle and lower thoracic esophageal cancer underwent Ivor-Lewis esophagectomy, and patients with upper thoracic esophageal cancer underwent Akiyama esophagectomy. 2-field (thoracic and abdominal) lymph node metastases information and the 1, 3, 5, 10-year survival rates were analyzed retrospectively. RESULTS: Lymph node metastases were found in 713 patients. The lymph node metastases rate was 42.2% (713/1690).Thoracic lymph node metastasis rate was 39.3% (665/1690), among which in the right pleural apical para-tracheal triangle was 20.7% (349/1690), in the posterior upper mediastinum was 26.3% (444/1690), in the lower mediastinum was 18.2% (307/1690). Abdominal lymph node metastasis rate was 20.1% (339/1690). THE Postoperative complication rate was 16.4% (278/1690), among which the pulmonary complication rate ranking the first, was 43.6% (136/312). The operative mortality rate was 0.2%. The 1-year, 3-year, 5-year and 10-year survival rates were 88.2% (1388/1574), 63.5% (868/1367), 54.8% (705/1287) and 30.8% (232/754), respectively. The 5-year survival rate in patients without lymph node metastasis was 76.2% (448/588), but that in patients with lymph node metastases was 36.8% (257/669). CONCLUSION: The results of this study demonstrated that Ivor-Lewis and Akiyama esophagectomy with two-field lymph node dissection exposes the operation fields clearly and make radical lymphadenectomy thoroughly, especially the lymph nodes in the posterior upper mediastinum around the recurrent laryngeal nerve and in the right pleural apical para-tracheal triangle. It is essential that patients with esophageal carcinoma with lymph node metastases should undergo esophagectomy with extended 2-field dissection of lymph nodes. This can elevate the postoperative 5-year survival rate remarkably.

[Regulations and lymphadenectomy strategy of mediastinal and upper abdominal lymph node metastasis in thoracic esophageal carcinoma].

BACKGROUND & OBJECTIVE: Regional lymph node metastasis plays an important role in the prognosis of esophageal carcinoma. However, the range of lymph node dissection is still controversial. This study was to investigate the regulations of lymph node metastasis of thoracic esophageal carcinoma in the mediastinum and upper abdomen, and explore the rational lymphadenectomy with Ivor-Lewis procedure. METHODS: A total of 1 412 thoracic esophageal carcinoma patients underwent radical esophagectomy and mediastinal and abdominal lymphadenectomy by Ivor-Lewis procedure from 1990 to 2005 at Yangzhong People's Hospital; 517 of them underwent right para-trachea triangle field lymphadenectomy through the right pleural apical approach. The regulations of regional lymph node metastasis were analyzed. RESULTS: Of the 1,412 patients, 323 (22.88%) had postoperative complications, 2 (0.14%) died during hospitalization, and 547 (38.74%) had lymph node metastasis. The lymph node metastasis rates were 32.30% in the right para-trachea triangle, 18.43% in the upper mediastinum, 5.31% in the lower mediastinum, and 17.28% in the upper abdomen(P<0.001). Of the 13 916 resected lymph nodes, 2 662 (19.13%) were positive; the metastasis degree (positive lymph nodes/resected lymph modes) were 23.83% in the right para-trachea triangle, 18.92% in the upper mediastinum, 21.07% in the lower mediastinum, and 17.20% in the upper abdomen. For those patients with the cancer focuses in the upper, middle and lower segments of the esophagus, the lymph node metastasis rates were 40.59%, 36.97% and 44.35% (P=0.093), respectively, while the lymph node metastasis degree in these 3 fields were 19.60%, 18.35%, and 21.82%, respectively. Both the lymph node metastasis rate and degree were significantly higher in the patients at advanced stage than in the patients at early stage (46.56% vs. 7.75%, 21.82% vs. 4.01%, P<0.001). CONCLUSIONS: Regional lymph node metastasis, especially in the right para-trachea triangle and upper mediastinum, is a key factor for thoracic esophageal carcinoma. Ivor-Lewis esophagectomy with two-field lymph node dissection is a safe operation for thoracic esophageal carcinoma, and may increase the chances of complete resection.

[Right para-tracheal triangle lymphadenectomy for esophageal carcinoma].

OBJECTIVE: To investigate the incidence of lymph node metastasis (LNM) in the right para-tracheal triangle (RPT) of esophageal carcinoma patients and the technique of dissection. METHODS: On the top of double mediastinal and abdominal lymphadenectomy, 333 esophageal carcinoma patients received RPT lymphadenectomy through the right pleural apical approach from 1990 to 2001. RESULTS: In these 333 patients, the lymph node metastasis (LNM) rate in the RPT was 36.40%. A total of 457 nodes among 2 159 nodes removed gave a metastasis degree of 24.96%. The LNM rates in RPT for cervical, upper third, middle third, and lower third segments of esophagus were 66.67%, 45.45%, 34.19% and 15.79% (P < 0.05), while their respective metastasis degrees were 44.44%, 27.04%, 24.32% and 18.92% (P > 0.05). The frequency of positive nodes in the RPT for PTI, PT1, PT2, PT3 and PT4 was 0, 17.24%, 28.7%, 45.16% and 53.57%, while those of metastasis degree were 0, 8.77%, 17.62%, 33% and 41.17% (P < 0.01). The frequency of LNM in the RPT in papillary, erosive, patch-like and covert type of early tumor was 40%, 3.85%, 0 and 0 (P < 0.05), while those of the metastasis degree were 29.41%, 1.82%, 0 and 0 (P < 0.01). Higher rate of LNM in progressive stenotic esophageal carcinoma was observed compared with those of the other gross types (56.52%, P < 0.05), so was the degree (P < 0.01). The frequency of LNM in the RPT for mono-focal and multi-focal tumor was 34.98% and 70% without significant difference (P > 0.05), while the degree was 24.29% and 53.33% (P < 0.05). Postoperative complications were: leak (0.6%), and recurrent laryngeal nerve injury (1.2%). No injury of vein or infra-clavicular artery, tracheal damage or mortality occurred. CONCLUSION: 1. The lymph node metastasis from esophageal carcinoma has a tendency of wide spread and right para-tracheal triangle is an important region to be doomed. 2. With location, depth of tumor invasion and differentiation of tumor as major factors affecting LNM of esophageal carcinoma, dissection of this region should be paid more emphasis. 3. In early lesions, higher frequency of LNM in the RPT is found in papillary and erosive lesions than in the other macroscopic types. 4. Exposing the RPT, lymph node by dissection through a right pleural apical approach is very important and significant.

[Effect of moulding and extruding conditions on mechanical properties of poly(D,L-lactide) and MDI chain-extending poly(D,L-lactide)/hydroxyapatite composite].

In this paper, poly(D,L-lactide) (PDLLA), MDI chain-extending poly(D,L-lactide) (PDLLA/MDI) and MDI chain-extending poly(D,L-lactide)/hydroxyapatite composite (PDLLA/HA/MDI) were prepared respectively and the effects of moulding and extruding conditions on their mechanical properties were also investigated. At the optimal conditions, bending strength of PDLLA and PDLLA/MDI is 35.1 MPa and 51.3 MPa, respectively, and their bending modulus is 2413.6 MPa and 1830.9 MPa, respectively. Bending strength of PDLLA/HA and PDLLA/HA/MDI is 31.2 MPa and 55.4 MPa, respectively, and their bending modulus is 1735.0 MPa and 2068.5 MPa, respectively. These results have shown that the mechanical properties of PDLLA/MDI and PDLLA/HA/MDI have enhanced significantly by MDI chain-extending.