CXCL8 Knockout: A Key to Resisting Pasteurella multocida Toxin-Induced Cytotoxicity.

Pasteurella multocida, a zoonotic pathogen that produces a 146-kDa modular toxin (PMT), causes progressive atrophic rhinitis with severe turbinate bone degradation in pigs. However, its mechanism of cytotoxicity remains unclear. In this study, we expressed PMT, purified it in a prokaryotic expression system, and found that it killed PK15 cells. The host factor CXCL8 was significantly upregulated among the differentially expressed genes in a transcriptome sequencing analysis and qPCR verification. We constructed a CXCL8-knockout cell line with a CRISPR/Cas9 system and found that CXCL8 knockout significantly increased resistance to PMT-induced cell apoptosis. CXCL8 knockout impaired the cleavage efficiency of apoptosis-related proteins, including Caspase3, Caspase8, and PARP1, as demonstrated with Western blot. In conclusion, these findings establish that CXCL8 facilitates PMT-induced PK15 cell death, which involves apoptotic pathways; this observation documents that CXCL8 plays a key role in PMT-induced PK15 cell death.

The prognostic value of preoperative systemic inflammatory response index in predicting outcomes of acute type A aortic dissection patients underwent surgical treatment.

Background: The systemic inflammatory response index (SIRI) is a novel inflammatory-immune biological marker that has prognostic value in various cardiovascular diseases. This study aims to investigate the relationship between SIRI and short-term and long-term prognosis in patients with acute type A aortic dissection (AAAD) underwent surgical treatment. Methods: We conducted a retrospective analysis of patients with AAAD who underwent emergency surgical treatment at our center. Through multifactorial logistics regression analysis and cox proportional hazards regression analysis, we identified SIRI as an independent risk factor for major adverse events (MAEs) and long-term aorta-related adverse events (ARAEs) post-surgery. The optimal cutoff value of preoperative SIRI was determined using receiver operating characteristic (ROC) curve analysis, and patients were divided into low SIRI group and high SIRI group. The prognostic outcomes at different time points post-surgery for the two groups of patients were analyzed using Kaplan-Meier survival analysis, and the significance was determined by log-rank test. Results: A total of 691 AAAD patients were included in this study. Among them, 50 patients (7.2%) died within 30 days post-surgery, and 175 patients (25.3%) experienced MAEs. A total of 641 patients were followed up, with an average follow-up time of 33.5 ± 17.5 months, during which 113 patients (17.6%) experienced ARAEs. The results of multifactorial logistics regression analysis and cox proportional hazards regression analysis showed that SIRI was an independent risk factor for postoperative MAEs (OR=3.148, 95%CI[1.650-6.006], p<0.001) and ARAEs (HR=2.248, 95%CI[1.050-4.809], p<0.037). Kaplan-Meier analysis demonstrated that the MAEs-free survival in the high SIRI group was significantly lower than that in the low SIRI group, and a similar trend was observed in the ARAEs-free survival during follow-up (log-rank test, p<0.001). Conclusion: Preoperative SIRI is significantly associated with the short-term and long-term prognosis of AAAD patients underwent emergency open surgery, demonstrating its valuable prognostic value. Therefore, preoperative SIRI is a reliable biological marker that can serve as a valuable tool for preoperative risk stratification and decision management.

Dapagliflozin prevents kidney podocytes pyroptosis via miR-155-5p/HO-1/NLRP3 axis modulation.

Diabetic nephropathy (DN) is a significant clinical microvascular complication associated with diabetes mellitus (DM), and end-stage diabetes giving rise to kidney failure is developing into the major etiological factor of chronic kidney failure. Dapagliflozin is reported to limit podocyte damage in DM, which has proven to protect against renal failure. Mounting evidence has demonstrated that pyroptosis is associated with DM progression. Nevertheless, whether pyroptosis causes DN and the underlying molecular pathways remain obscure. In this study, we aimed to explore the antipyroptotic attributes of dapagliflozin and elucidate the underlying mechanisms of kidney damage in diabetes. In vivo, experiments were conducted in streptozotocin (STZ)-induced type 2 diabetic mice, which were administered dapagliflozin via gavage for 6 weeks. Subsequently, the specific organizational characteristics and expression of pyroptosis-related genes were evaluated. Intragastric dapagliflozin administration markedly reduced renal tissue injury. Meanwhile, dapagliflozin also attenuated the expression level of pyroptosis associated genes, including ASC, cleaved Caspase-1, GSDMD N-termini, NLRP3, IL-18, and IL-1ß in renal tissue of dapagliflozin-treated animals. Similar antipyroptotic effects were observed in palmitic acid (PA)-treated mouse podocytes. We also found that heme oxygenase 1 (HO-1) enhanced the protection of mouse podocyte clone 5 cells (MPC5). Moreover, miR-155-5p inhibition increased pyroptosis in PA-treated MPC5 cells, suggesting that miR-155-5p acts as an endogenous stimulator that increases HO-1 expression and reduces pyroptosis. Hence, our findings imply that dapagliflozin inhibits podocyte pyroptosis via the miR-155-5p/HO-1/NLRP3 axis in DM. Furthermore, dapagliflozin substitution may be regarded as an effective strategy for preventing pyroptosis in the kidney, including a therapeutic option for treating pyroptosis-related DN.

Application of neoadjuvant chemoradiotherapy and neoadjuvant chemotherapy in curative surgery for esophageal cancer: A meta-analysis.

BACKGROUND: The effectiveness of neoadjuvant therapy in esophageal cancer (EC) treatment is still a subject of debate. AIM: To compare the clinical efficacy and toxic side effects between neoadjuvant chemoradiotherapy (nCRT) and neoadjuvant chemotherapy (nCT) for locally advanced EC (LAEC). METHODS: A comprehensive search was conducted using multiple databases, including PubMed, EMBASE, MEDLINE, Science Direct, The Cochrane Library, China National Knowledge Infrastructure, Wanfang Database, Chinese Science and Technology Journal Database, and Chinese Biomedical Literature Database Article. Studies up to December 2022 comparing nCRT and nCT in patients with EC were selected. RESULTS: The analysis revealed significant differences between nCRT and nCT in terms of disease-free survival. The results indicated that nCRT provided better outcomes in terms of the 3-year overall survival rate (OSR) [odds ratio (OR) = 0.95], complete response rate (OR = 3.15), and R0 clearance rate (CR) (OR = 2.25). However, nCT demonstrated a better 5-year OSR (OR = 1.02) than nCRT. Moreover, when compared to nCRT, nCT showed reduced risks of cardiac complications (OR = 1.15) and pulmonary complications (OR = 1.30). CONCLUSION: Overall, both nCRT and nCT were effective in terms of survival outcomes for LAEC. However, nCT exhibited better performance in terms of postoperative complications.

Prediction of pathological complete response of breast cancer patients who received neoadjuvant chemotherapy with a nomogram based on clinicopathologic variables, ultrasound, and MRI.

OBJECTIVE: To establish a nomogram for predicting the pathologic complete response (pCR) in breast cancer (BC) patients after NAC by applying magnetic resonance imaging (MRI) and ultrasound (US). METHODS: A total of 607 LABC women who underwent NAC before surgery between January 2016 and June 2022 were retrospectively enrolled, and then were randomly divided into the training (n = 425) and test set (n = 182) with the ratio of 7:3. MRI and US variables were collected before and after NAC, as well as the clinicopathologic features. Univariate and multivariate logistic regression analyses were applied to confirm the potentially associated predictors of pCR. Finally, a nomogram was developed in the training set with its performance evaluated by the area under the receiver operating characteristics curve (ROC) and validated in the test set. RESULTS: Of the 607 patients, 108 (25.4%) achieved pCR. Hormone receptor negativity (odds ratio [OR], 0.3; P < .001), human epidermal growth factor receptor 2 positivity (OR, 2.7; P = .001), small tumour size at post-NAC US (OR, 1.0; P = .031), tumour size reduction ≥50% at MRI (OR, 9.8; P < .001), absence of enhancement in the tumour bed at post-NAC MRI (OR, 8.1; P = .003), and the increase of ADC value after NAC (OR, 0.3; P = .035) were all significantly associated with pCR. Incorporating the above variables, the nomogram showed a satisfactory performance with an AUC of 0.884. CONCLUSION: A nomogram including clinicopathologic variables and MRI and US characteristics shows preferable performance in predicting pCR. ADVANCES IN KNOWLEDGE: A nomogram incorporating MRI and US with clinicopathologic variables was developed to provide a brief and concise approach in predicting pCR to assist clinicians in making treatment decisions early.

Porcine reproductive and respiratory syndrome virus upregulates SMPDL3B to promote viral replication by modulating lipid metabolism.

Porcine reproductive and respiratory syndrome virus (PRRSV) poses a severe threat to the health of pigs globally. Host factors play a critical role in PRRSV replication. Using PRRSV as a model for genome-scale CRISPR knockout (KO) screening, we identified a host factor critical to PRRSV infection: sphingomyelin phosphodiesterase acid-like 3B (SMPDL3B). Our findings show that SMPDL3B restricted PRRSV attachment, entry, replication, and secretion and that its depletion significantly inhibited PRRSV proliferation, indicating that SMPDL3B plays a positive role in PRRSV replication. Our data also show that SMPDL3B deficiency resulted in an accumulation of intracellular lipid droplets (LDs). The expression level of key genes (ACC, SCD-1, and FASN) involved in lipogenesis was increased, whereas the fundamental lipolysis gene, ATGL, was inhibited when SMPDL3B was knocked down. Overall, our findings suggest that SMPDL3B deficiency can effectively inhibit viral infection through the modulation of lipid metabolism.

Immunogenicity and protective efficacy of a multi-epitope recombinant toxin antigen of Pasteurella multocida against virulent challenge in mice.

Pasteurella multocida (P. multocida) infection frequently results in porcine atrophic rhinitis and swine plague, leading to large economic losses for the swine industry worldwide. P. multocida toxin (PMT, 146 kDa) is a highly virulent key virulence factor that plays a vital role in causing lung and turbinate lesions. This study developed a multi-epitope recombinant antigen of PMT (rPMT) that showed excellent immunogenicity and protection in a mouse model. Using bioinformatics to analyse the dominant epitopes of PMT, we constructed and synthesized rPMT containing 10 B-cell epitopes, 8 peptides with multiple B-cell epitopes and 13 T-cell epitopes of PMT and a rpmt gene (1,974 bp) with multiple epitopes. The rPMT protein (97 kDa) was soluble and contained a GST tag protein. Immunization of mice with rPMT stimulated significantly elevated serum IgG titres and splenocyte proliferation, and serum IFN-γ and IL-12 were upregulated by 5-fold and 1.6-fold, respectively, but IL-4 was not. Furthermore, the rPMT immunization group exhibited alleviated lung tissue lesions and a significantly decreased degree of neutrophil infiltration compared with the control groups post-challenge. In the rPMT vaccination group, 57.1% (8/14) of the mice survived the challenge, similar to the bacterin HN06 group, while all the mice in the control groups succumbed to the challenge. Thus, rPMT could be a suitable candidate antigen for developing a subunit vaccine against toxigenic P. multocida infection.

Pseudorabies virus exploits N6-methyladenosine modification to promote viral replication.

Introduction: Pseudorabies virus (PRV) is the pathogenic virus of porcine pseudorabies (PR), belonging to the Herpesviridae family. PRV has a wide range of hosts and in recent years has also been reported to infect humans. N6-methyladenosine (m6A) modification is the major pathway of RNA post-transcriptional modification. Whether m6A modification participates in the regulation of PRV replication is unknown. Methods: Here, we investigated that the m6A modification was abundant in the PRV transcripts and PRV infection affected the epitranscriptome of host cells. Knockdown of cellular m6A methyltransferases METTL3 and METTL14 and the specific binding proteins YTHDF2 and YTHDF3 inhibited PRV replication, while silencing of demethylase ALKBH5 promoted PRV output. The overexpression of METTL14 induced more efficient virus proliferation in PRV-infected PK15 cells. Inhibition of m6A modification by 3-deazaadenosine (3-DAA), a m6A modification inhibitor, could significantly reduce viral replication. Results and Discussion: Taken together, m6A modification played a positive role in the regulation of PRV replication and gene expression. Our research revealed m6A modification sites in PRV transcripts and determined that m6A modification dynamically mediated the interaction between PRV and host.

Inhibiting von Hippel‒Lindau protein-mediated Dishevelled ubiquitination protects against experimental parkinsonism.

Dopaminergic neuron degeneration is a hallmark of Parkinson's disease (PD). We previously reported that the inactivation of von Hippel‒Lindau (VHL) alleviated dopaminergic neuron degeneration in a C. elegans model. In this study, we investigated the specific effects of VHL loss and the underlying mechanisms in mammalian PD models. For in vivo genetic inhibition of VHL, AAV-Vhl-shRNA was injected into mouse lateral ventricles. Thirty days later, the mice received MPTP for 5 days to induce PD. Behavioral experiments were conducted on D1, D3, D7, D14 and D21 after the last injection, and the mice were sacrificed on D22. We showed that knockdown of VHL in mice significantly alleviated PD-like syndromes detected in behavioral and biochemical assays. Inhibiting VHL exerted similar protective effects in MPP+-treated differentiated SH-SY5Y cells and the MPP+-induced C. elegans PD model. We further demonstrated that VHL loss-induced protection against experimental parkinsonism was independent of hypoxia-inducible factor and identified the Dishevelled-2 (DVL-2)/ß-catenin axis as the target of VHL, which was evolutionarily conserved in both C. elegans and mammals. Inhibiting the function of VHL promoted the stability of ß-catenin by reducing the ubiquitination and degradation of DVL-2. Thus, in vivo overexpression of DVL-2, mimicking VHL inactivation, protected against PD. We designed a competing peptide, Tat-DDF-2, to inhibit the interaction between VHL and DVL-2, which exhibited pharmacological potential for protection against PD in vitro and in vivo. We propose the therapeutic potential of targeting the interaction between VHL and DVL-2, which may represent a strategy to alleviate neurodegeneration associated with PD.

Symbiotic microbiota and odor ensure mating in time for giant pandas.

To achieve reproduction, male solitary mammals need to locate females using chemical communication with high levels of precision. In the case of giant pandas, the total estrus period of females was usually 15 days each year, however, successful mating activity is finished within 3 days from respective home range. The mating pattern of giant pandas, where multiple males compete for each female requires females employ efficient systems to communicate their estrus phases. To verifying whether the scent secretions of giant pandas changes by gender and estrus progression, the microbiota and compounds in 29 anogenital gland samples from 14 individuals during estrus were analyzed by 16S rRNA sequencing and GC-MS. We show that the microbiota communities covary by gender with 4 particular compounds of scent secretions. Among 597 genera, 34 were identified as biomarkers that could be used to distinguish between different estrus phases. By bacterial-compounds co-analysis, 3 fatty ester acids and squalene compounds covaried with the development of estrus in the bacterial communities of female giant pandas. This study helps clarify how a large, solitary mammal expresses accurate information to improve the likelihood of successful reproduction by changing the composition of microbiota and odor compounds of anogenital glands during estrus.

Efficacy of traditional Chinese medicine combined with rivaroxaban in the treatment of lower extremity deep vein thrombosis: A meta-analysis.

BACKGROUND: Despite the usefulness of traditional Chinese medicine (TCM) in the treatment of lower deep vein thrombosis (DVT), there is no consensus on safety and efficacy. We aim to systematically evaluate the safety and efficacy of TCM combined with Rivaroxaban in the treatment of lower limb DVT. METHODS: An online search of databases such as Cochrane Library, Embase, Pubmed, and Web of science, as well as CBM, China Science and Technology Journal Database, China Knowledge Network (CNKI) and Wanfang Data (from inception to July, 2021) was performed. All published clinical randomized controlled trials (RCTs) were screened manually, evaluated for quality and considered for meta-analysis using RevMan 5.3. RESULTS: Nine RCTs with a total of 730 cases were included, 368 cases in the trial group were treated with TCM combined with Rivaroxaban, and 362 cases in the control group were treated with Rivaroxaban alone after surgery. Clinical efficiency was significantly higher in the test group [OR = 3.33, 95% CI (2.01, 5.53), P < .00001], the circumference of the affected limb was significantly lower in the thigh and calf, respectively [MD = -1.48, 95% CI (-1.88, -1.09), P < .00001], [MD = -0.54, 95% CI (-0.62, -0.46), P < .00001], pain scores were significantly lower [MD = -0.97, 95% CI (-1.58, -0.36), P = .002], coagulation index plasma fibrinogen (FIB) was significantly lower [MD = -0.85, 95% CI (-1.18, -0.52), P < .00001], coagulation function index D-2 aggregates were significantly reduced [MD = -0.69, 95% CI (-1.13, -0.24), P = .002], serum hypersensitive C-reactive protein (hs-CRP) measurements were significantly reduced [MD = -5.37, 95% CI (-9.20, -1.55), P = .006], complications measurement was significantly lower [OR = 0.60, 95% CI (0.27, 1.30), P = .20], activated partial thrombin time (ATPP) measurement was significantly lower [MD = 5.70, 95% CI (4.28, 7.12), P < .00001] and prothrombin time (PT) measurement was significantly lower [MD = 1.64, 95% CI (0.70, 2.57), P = .0006]. CONCLUSION: Based on the available evidence, TCM combined with Rivaroxaban for treating lower extremity DVT have better clinical efficacy and safety profile, reducing the risk of bleeding complications and adverse effects. Further improved studies are needed to support this inference.

[Responses of radial growth of Pinus tabuliformis to climate change at the northern slopes of Qinling Mountains, China]. / 秦岭北麓油松径向生长对气候变化的响应.

A standard chronology (STD) was established with Pinus tabuliformis samples collected from Nanwutai at the northern slopes of the Qinling Mountains. The correlations between radial growth of P. tabuliformis and climatic factors were explored. The results showed that radial growth was positively correlated with moisture factors in previous September and current May. The radial growth was positively correlated with temperature factors in pre-vious November and negatively with that in previous October and current May. There was significant lag effect of climatic factors on the radial growth. The radial growth of P. tabuliformis was well correlated with the PDSI drought severity index, with positive correlations being identified from September to December of the previous year and in May of the current year. The regression model was able to simulate the relationship between STD and PDSI. The formations of extremely wide and narrow tree-ring were mainly a result of drought condition. Combined with climatic factors, PDSI could better reflect the radial growth characteristics of P. tabuliformis.

Diagnostic performance of DCE-MRI, multiparametric MRI and multimodality imaging for discrimination of breast non-mass-like enhancement lesions.

OBJECTIVE: The aim of this study was to investigate and compare the diagnostic performance of dynamic contrast-enhanced (DCE)-MRI, multiparametric MRI (mpMRI), and multimodality imaging (MMI) combining mpMRI and mammography (MG) for discriminating breast non-mass-like enhancement (NME) lesions. METHODS: This retrospective study enrolled 193 patients with 199 lesions who underwent 3.0 T MRI and MG from January 2017 to December 2019. The features of DCE-MRI, turbo inversion recovery magnitude (TIRM), and diffusion-weighted imaging (DWI) were assessed by two breast radiologists. Then, all lesions were divided into microcalcification and non-microcalcification groups to assess the features of MG. Comparisons were performed between groups using univariate analyses. Then, multivariate analyses were performed to construct diagnostic models for distinguishing NME lesions. Diagnostic performance was evaluated by using the area under the curve (AUC) and the differences between AUCs were evaluated by using the DeLong test. RESULTS: Overall (n = 199), mpMRI outperformed DCE-MRI alone (AUCmpMRI = 0.924 vs. AUCDCE-MRI = 0.884; p = 0.007). Furthermore, MMI outperformed both mpMRI and MG (the microcalcification group [n = 140]: AUCMMI = 0.997 vs. AUCmpMRI = 0.978, p = 0.018 and AUCMMI = 0.997 vs. AUCMG = 0.912, p < 0.001; the non-microcalcification group [n = 59]: AUCMMI = 0.857 vs. AUCmpMRI = 0.768, p = 0.044 and AUCMMI = 0.857 vs. AUCMG = 0.759, p = 0.039). CONCLUSION & ADVANCES IN KNOWLEDGE: DCE-MRI combined with DWI and TIRM information could improve the diagnostic performance for discriminating NME lesions compared with DCE-MRI alone. Furthermore, MMI combining mpMRI and MG showed better discrimination than both mpMRI and MG.

The Plaque Microbiota Community of Giant Panda (Ailuropoda melanoleuca) Cubs With Dental Caries.

Dental caries severely hinders efficient access to adequate energy in wildlife. Different food supplies will develop characteristic plaque, and the microorganisms of these plaque are closely related to dental health. Here, plaque samples from panda cubs with caries and caries-free were collected for 16S rRNA high-throughput sequencing. All sequences clustered into 337 operational taxonomic units (OTUs; 97% identity), representing 268 independent species belonging to 189 genera, 98 families, 51 orders, 24 classes, and 13 phyla. Two groups shared 218 OTUs, indicating the presence of a core plaque microbiome. α diversity analysis showed that the microbial diversity in plaques with caries exceeded that of caries-free. The dominant phyla of plaque microbiota included Proteobacteria, Bacteroidetes, Firmicutes, Fusobacteria, and Actinobacteria. The dominant genera included unclassified Neisseriaceae, Actinobacillus, Lautropia, Neisseria, Porhyromonas, unclassified Pasteurellaceae, Moraxella, Streptococcus, Bergeywlla and Capnocytophaga. ß diversity analysis showed that the plaque microbial community structure was different between two groups. Using LEfSe analysis, 19 differentially abundant taxa were identified as potential biomarkers. Finally, function predictions analysis showed All the energy related metabolic pathways on KEGG level 2 were enriched in caries-active group. Consistent with the mainstream caries-causing narrative, our results illuminate the lack of information regarding the oral microflora composition and function within giant panda cubs.

Changes in the Mitochondria-Related Nuclear Gene Expression Profile during Human Oocyte Maturation by the IVM Technique.

To address which mitochondria-related nuclear differentially expressed genes (DEGs) and related pathways are altered during human oocyte maturation, single-cell analysis was performed in three oocyte states: in vivo matured (M-IVO), in vitro matured (M-IVT), and failed to mature in vitro (IM-IVT). There were 691 DEGs and 16 mitochondria-related DEGs in the comparison of M-IVT vs. IM-IVT oocytes, and 2281 DEGs and 160 mitochondria-related DEGs in the comparison of M-IVT vs. M-IVO oocytes, respectively. The GO and KEGG analyses showed that most of them were involved in pathways such as oxidative phosphorylation, pyruvate metabolism, peroxisome, and amino acid metabolism, i.e., valine, leucine, isoleucine, glycine, serine, and threonine metabolism or degradation. During the progress of oocyte maturation, the metabolic pathway, which derives the main source of ATP, shifted from glucose metabolism to pyruvate and fatty acid oxidation in order to maintain a low level of damaging reactive oxygen species (ROS) production. Although the immature oocytes could be cultured to a mature stage by an in vitro technique (IVM), there were still some differences in mitochondria-related regulations, which showed that the mitochondria were regulated by nuclear genes to compensate for their developmental needs. Meanwhile, the results indicated that the current IVM culture medium should be optimized to compensate for the special need for further development according to this disclosure, as it was a latent strategy to improve the effectiveness of the IVM procedure.

Coumaroyl and feruloyl flavonoid glycosides from the male flowers of Ginkgo biloba L. and their inhibitory activity against α-glucosidase.

Four flavonoid glycosides containing coumaroyl or feruloyl groups were isolated from the male flowers of Ginkgo biloba L., and compounds 3 and 4 were identified as novel compounds. The inhibitory activities against α-glucosidase were investigated by docking studies, in vitro assays and kinetic studies. The docking results showed that all compounds mainly formed hydrogen-bond and π-π-stacking interactions with α-glucosidase. Compound 4 had the lowest binding energy and maximum number of hydrogen bonds. Subsequently, the in vitro assays showed that compound 4 exhibited the strongest inhibitory potency. Finally, the kinetic studies indicated the inhibitory mode of compounds 1-4 against α-glucosidase were mixed types of competitive and non-competitive. Together, these findings suggested that the isolated flavonoid glycosides in this study, especially compound 4, have potential as α-glucosidase inhibitors.

Mechanism of Dendrobium huoshanense in Treatment of Gastric Ulcer： Based on Network Pharmacology and in Vivo Experiment / 中国实验方剂学杂志

ObjectiveTo explore the mechanism of Dendrobium huoshanense in the treatment of gastric ulcer （GU） based on network pharmacology and in vivo experiment. MethodThe active components of D. huoshanense were searched from Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP） and literature， and the targets of the components were screened from TCMSP and SwissTargetPrediction. GU-related genes were retrieved from GeneCards， Online Mendelian Inheritance in Man （OMIM）， and DisGeNET. Thereby， the common targets of the disease and the medicinal were yielded and the protein-protein interaction （PPI） network was constructed， followed by Gene Ontology （GO） term enrichment and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment. According to the predicted results， hematoxylin-eosin （HE） staining， immunohistochemistry， enzyme-linked immunosorbent assay （ELISA）， Western blot， and real-time fluorescent quantitative polymerase chain reaction （Real-time PCR） were used to validate the effects of D. huoshanense on acetic acid-induced GU in rats. ResultA total of 63 active components of D. huoshanense and 37 target genes of D. huoshanense for the treatment of GU were screened out. PPI network analysis yielded several possible core anti-GU targets of D. huoshanense. They influenced the development of GU by acting on signaling pathways such as phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt）， hypoxia inducible factor-1 （HIF-1）， tumor necrosis factor （TNF）， and nuclear factor-κB （NF-κB）， and various biological processes. The in vivo experiment showed that D. huoshanense significantly reduced the levels of inflammatory factors such as interleukin-1β （IL-1β）， interleukin-6 （IL-6）， and TNF-α in the serum of model rats （P<0.05， P<0.01）， increased gastric blood flow （GBF） at the ulcer margin， raised the expression of epidermal growth factor （EGF） and epidermal growth factor receptor （EGFR） at the ulcer margin （P<0.01）， significantly down-regulated protein and mRNA expression of PI3K and Akt， and up-regulated protein and mRNA expression of phosphatase and tensin homolog deleted on chromosome ten （PTEN） in the gastric tissues of GU rats （P<0.01）. ConclusionThrough regulating EGFR/PI3K/Akt signaling pathway， D. huoshanense can inhibit tissue inflammation， increase gastric microcirculatory blood flow at the ulcer margin， and promote cell proliferation and repair of damaged gastric mucosa.

Effect of extract of livistona chinensis on the proliferation and apoptosis of bladder cancer cells and its related mechanisms / 中华泌尿外科杂志

Objective:To study the effect of extract of livistona chinensis on the proliferation and apoptosis of bladder cancer cells and the related mechanism.Methods:T24 cells were cultured in medium with the final concentration of 0, 25, 50 and 100 mg/L livistona chinensis extract, respectively. And then they were divided into control group and low, medium and high dose groups. The cell survival rate was detected by cell counting kit 8 (CCK-8). Colony formation assay was used to detect the number of cell clones. Apoptosis was detected by flow cytometry. Western blot was used to detect the expression of related proteins. The Syk overexpression vector plasmid and its negative control were transfected into T24 cells. After transfection, the cells were treated with 100 mg/L livistona chinensis. The cell survival rate, colony formation number and apoptosis rate were detected by the above method. The bladder cancer model nude mice were treated with different concentrations of livistona chinensis extract. Under the microscope, the expression of protein was detected by immunohistochemical staining of bladder tissue.Results:Compared with the control group, the survival rate of T24 cells in the low, medium and high dose livistona chinensis extract groups were significantly decreased [(88.50±3.65)%, (70.58±2.47)%, (48.90±2.37)% vs. (98. 25±4.26)%], and the number of clone formation decreased significantly [(101. 33±3.40), (84.00±2.94), (60.00±2.16) vs. (121.33±4.64) ], and the apoptosis rate was significantly increased [(11.45± 0.59)%, (17.71±0.64)%, (21.33±0.83)% vs. (7. 86±0.43)%]. The expression level of Ki-67 protein was significantly decreased, while the expression levels of Caspase3 and Syk protein were significantly increased in a concentration dependent manner ( P < 0.05). The cell survival rate of pcDNA3.1-Syk group was significantly lower than that of pcDNA3.1 group [(63.87±2.53)% vs. (98. 45±3.54)%], the number of clone formation decreased significantly [(74. 33±2.87) vs. (121.33±3.68)], and the apoptosis rate was significantly increased [(18.39±0.63)% vs. (7.89± 0.45)%] (all P<0.05). The cell survival rate in the high-dose group of livistona chinensis+ pcDNA3.1-Syk was significantly lower than that in the high-dose group of livistona chinenisi+ pcDNA3.1 group [ (29.80±1.63)% vs.(49.33±2.76)% ], the number of clone formation decreased significantly [(33.00±2.94) vs. (59.67±3.30) ], and the apoptosis rate was significantly increased [(26.93±0.68)% vs. (21.25±0.78)% ]( P<0.05). The experimental results of nude mice of bladder cancer model showed that the tumor volume of transplanted bladder cancer nude mice in the control group and the low, medium, and high dose livistona chinensis extract groups were (1 209.75±64.37), (1 006.31±40.49), (530.58±42.87), (267.58±16.73)mm 3, respectively, the weight of the transplanted tumor were (0.36±0.08), (0.30±0.04), (0.26±0.03), (0.18±0.06)g, and the differences between the two groups were statistically significant ( P <0.05). Immunohistochemical staining results showed that the expression of Sky and Caspase3 was increased and the expression of Ki-67 was decreased in the middle and high dose groups compared with that in the control group. Conclusion:Extract of livistona chinensis can inhibit the proliferation and promote apoptosis of bladder cancer cells by up regulating Syk expression.

Regulatory effect of m6 A modification on different viruses.

N6 -methyladenosine (m6 A) modification is the most common and reversible posttranscriptional modification of RNA in eukaryotes, which is mainly regulated by methyltransferase, demethylase, and specific binding protein. The replication of the virus and host immune response to the virus are affected by m6 A modification. In different kinds of viruses, m6 A modification has two completely opposite regulatory functions. This paper reviews the regulatory effects of m6 A modification on different viruses and provides a reference for studying the regulatory effects of RNA epitranscriptomic modification.

Molecular and functional characterization of HtrA protein in Actinobacillus pleuropneumoniae.

Actinobacillus pleuropneumoniae (A.pleuropneumoniae) causes serious economic loss for the swine industry. A high-temperature requirements A (HtrA)-like protease and its homologs have been reported to be involved in protein quality control and expression of important immunoprotective antigens in many pathogens. In this study, we showed that HtrA of A.pleuropneumoniae exhibited both chaperone and proteolytic activities. Moreover, Outer membrane protein P5 (OmpP5) in A.pleuropneumoniae and Heat shock protein 90 (Hsp90) in porcine lung tissues were first discovered and identified as specific proteolytic substrates for rHtrA. The maximum cleavage activity occurs at 50 ℃ in a time-dependent manner. In addition, rHtrA mainly induced IgG 2a subtype of IgG and Th1 (IFN-γ, IL-2) response in a mice model, and promoted a significant proliferation of spleen lymphocytes compare with negative control (P < 0.05). The survival rates of 37.5 % were observed against A.pleuropneumoniae strain. Together, these data demonstrate that rHtrA plays a multi-functional role in A.pleuropneumoniae.