Helicobacter pylori East Asian type CagA hijacks more SHIP2 by its EPIYA-D motif to potentiate the oncogenicity.

CagA is a significant oncogenic factor injected into host cells by Helicobacter pylori, which is divided into two subtypes: East Asian type (CagAE), characterized by the EPIYA-D motif, and western type (CagAW), harboring the EPIYA-C motif. CagAE has been reported to have higher carcinogenicity than CagAW, although the underlying reason is not fully understood. SHIP2 is an intracellular phosphatase that can be recruited by CagA to perturb the homeostasis of intracellular signaling pathways. In this study, we found that SHIP2 contributes to the higher oncogenicity of CagAE. Co-Immunoprecipitation and Pull-down assays showed that CagAE bind more SHIP2 than CagAW. Immunofluorescence staining showed that a higher amount of SHIP2 recruited by CagAE to the plasma membrane catalyzes the conversion of PI(3,4,5)P3 into PI(3,4)P2. This alteration causes higher activation of Akt signaling, which results in enhanced IL-8 secretion, migration, and invasion of the infected cells. SPR analysis showed that this stronger interaction between CagAE and SHIP2 stems from the higher affinity between the EPIYA-D motif of CagAE and the SH2 domain of SHIP2. Structural analysis revealed the crucial role of the Phe residue at the Y + 5 position in EPIYA-D. After mutating Phe of CagAE into Asp (the corresponding residue in the EPIYA-C motif) or Ala, the activation of downstream Akt signaling was reduced and the malignant transformation of infected cells was alleviated. These findings revealed that CagAE hijacks SHIP2 through its EPIYA-D motif to enhance its carcinogenicity, which provides a better understanding of the higher oncogenic risk of H. pylori CagAE.

Cognitive behavioral therapy achieves better benefits in relieving postoperative pain and improving joint function: A systematic review and meta-analysis of randomized controlled trials.

BACKGROUND: Cognitive behavioral therapy (CBT) is a structured, short-term psychotherapy approach that may have positive effects in terms of relieving postoperative pain. The main objective of this study was to determine the effect of CBT on pain and joint function in patients after total joint arthroplasty. METHODS: We searched 3 electronic databases including randomized controlled studies (RCTs) using CBT as an intervention. The main results of this study were to determine pain intensity by NRS, VAS, WOMAC pain Scale, PCS, and joint function by HHS, OKS, EQ-5D, ROM. Data extraction and quality assessment of included RCTs were independently performed by the authors and date analysis was performed by RevMan V.5.4. RESULTS: Among the 605 studies, 9 RCTS were included in this systematic review and meta-analysis. The study showed that the difference between CBT and usual care groups in PCS (≤3months), NRS, VAS (≤3months) were statistically significant (P < 0.05); the difference between CBT and usual care groups in PCS (≥12months), WOMAC Pain Scale, and VAS (≥12months) were not statistically significant (P > 0.05), indicating that CBT can improve pain in patients after arthroplasty in the early term. In addition, the difference between CBT and usual care groups in OKS (≤3months), HSS, ROM (≤3months), EQ-5D (≤3months) were not statistically significant (P > 0.05); the difference between CBT and usual care groups in EQ-5D (≥12months) were statistically significant (P < 0.05), indicating that the quality of life in patients after total joint arthroplasty were improved with the extension of follow-up time. CONCLUSIONS: This study shows that CBT can relieve pain in patients with total joint arthroplasty in the early postoperative period and improve quality of life to some extent over time.

Nuclear factor E2-associated factor 2 and musculoaponeurotic fibrosarcoma K mediate regulation glutathione peroxidase of Cristaria plicata after microcystin-induced oxidative stress.

Nuclear factor E2-associated factor 2 (Nrf2)/Antioxidant Response Element (ARE) signaling pathway is an endogenous antioxidant pathway that protects cells from oxidative damage. This pathway is triggered when aquatic organisms are exposed to environmental toxicants. In this study, CpMafK (musculoaponeurotic fibrosarcoma K of Cristaria plicata) mRNA expression in hepatopancreas and gills were up regulated after Cristaria plicata (C. plicata) was exposed to microcystin (MC), which showed that CpMafK protected C. plicata from MC. After MC treatment and CpNrf2 (Nrf2 of Cristaria plicata) knockdown, the mRNA expression of CpMafK was down regulated. After MC treatment and CpMafK knockdown, the mRNA expression of CpNrf2 was down regulated. Indicating that the expression of CpNrf2 was positively correlated with CpMafK. CpGPx (GPx of Cristaria plicata) mRNA was also down regulated with the down regulation of CpMafK and CpNrf2. CpGPx promoter contains a variety of transcription factor binding sites, including Nrf2, ARE elements, etc. Gel blocking experiments showed that CpNrf2/CpMafK heterodimers were bound to CpGPx promoters in vitro. Dual luciferase reporter assay showed that CpNrf2/CpMafK heterodimer negatively regulated CpGPx promoter in cells. In conclusion, Nrf2 and MafK mediate regulation of GPx play a crucial role in protecting bivalves from MC.

The thioredoxin expression of Cristaria plicata is regulated by Nrf2/ARE pathway under microcystin stimulation.

Thioredoxin plays an important role in inhibiting apoptosis and protecting cells from oxidative stress. This study was aimed to clarify how the expression of Trx from Cristaria plicata is regulated by Nrf2/ARE pathway. The expression of CpTrx mRNA was significantly up-regulated in gill and kidney tissues under microcystin stress. The Nrf2 gene of Cristaria plicata was identified to possess an auto active domain bit. While CpNrf2 was knocked down by specific small RNA, CpTrx mRNA expression was significantly down-regulated. The promoter of CpTrx gene had high transcriptional activity, and this basic transcriptional activity persisted after ARE element mutation. The region of promoter -206 to +217 bp was a core promoter region and had forward regulatory elements. Gel shift Assay exhibited that the CpTrx promoter could bind to the purified proteins CpNrf2 and CpMafK in vitro. The binding phenomenon disappeared after the ARE element mutation in promoter region. Subcellular localization experiments displayed that fluorescence overlap between CpNrf2 and Trx promoter increased under microcystin toxin stress. These results suggested that Trx expression was regulated by Nrf2/ARE pathway under oxidative stress.

Prx5 of Cristaria plicata has antioxidant function and is regulated by Nrf2/ARE signaling pathway.

Cristaria plicata is one of the more important freshwater pearl bivalves in China, which is susceptible to pathogen infection, and greatly impacts the ability of breeding pearls. Nrf2/ARE signaling pathway and its downstream target gene Prx5 have endogenous antioxidant functions to protect cells from oxidative damage. The full-length cDNA of Prx5 was cloned from C. Plicata, which was 1420 bp, encoding a total of 189 amino acids and had two conserved cysteine residues (Cys78 and Cys179). The amino acid sequence of CpPrx5 was highly similar to Prx5 of other species. Real-time fluorescence quantitative PCR showed that CpPrx5 was distributed in various tissues of mussels, and the highest expression was in hepatopancreas. The expression of CpPrx5 up-regulated in hepatopancreas and gills after LPS, PGN and Poly:I:C stimulation. The recombinant plasmid DE3-PGEX-4T-1-CpPrx5 was expressed in Escherichia coli BL21 and showed antioxidant activity. With the increase of CpPrx5 protein concentration, the superhelical form of DNA was protected. The expression of CpPrx5 was up-regulated after interference CpKeap1 and down-regulated after interference CpNrf2. Gel block assay showed that CpNrf2 and CpMafK proteins blocked CpPrx5 promoter. Subcellular localization showed that CpPrx5 was located in 293T nucleus and cytoplasm and CpMafK was located in 293T nucleus. GST-Pull down verified that CpMafK and CpPrx5 could bind in vitro. These results indicated that Prx5 had antioxidant function and could protects DNA from oxidative damage, and participated in transcriptional regulation by combining with the transcription factor MafK. In addition, MafK could combine with Nrf2 to regulate the downstream target gene Prx5.

Effect of Autochthonous Lactic Acid Bacteria-Enhanced Fermentation on the Quality of Suancai.

The lactic acid bacteria (LABs) used for fermentation have an extremely vital impact on the quality of Suancai, a fermented vegetable. The bacterial diversity and metabolites of inoculated Suancai with LABs, including Lactiplantibacillus plantarum (Lb. plantarum), Levilactabacillus brevis (Lb. brevis), and Leuconostoc mesenteroides (Leu. mesenteroides), were investigated. The inoculation of LABs significantly decreased the pH and the content of nitrite. The Suancai inoculated with LABs had a higher content of the total titratable acidity (TTA) and organic acids than spontaneous fermentation. The LABs inoculation significantly influenced the bacterial community structures, which directly or indirectly caused changes of metabolites. The bacterial community profiles of Suancai inoculated with Lb. plantarum were more similar to spontaneous fermentation. The inoculation of Lb. plantarum, Lb. brevis, and Leu. mesenteroides could increase its abundance in Suancai. Whatever the species inoculated, Lb. plantarum was always the predominant bacterium in Suancai after fermentation. The inoculated LABs were positively correlated with most volatile compounds and amino acids. The inoculated LABs significantly improved the volatile compounds and amino acid content of Suancai. This study could contribute to understanding the function of starters in Suancai fermentation and promote the selection of applicable starters for high-quality Suancai production.

Monitoring Mycoplasma pneumoniae-Specific Antibody, C-Reactive Protein, and Procalcitonin Levels in Children with Mycoplasma Pneumonia Is Important.

The goal of this study was to see how important it is to monitor Mycoplasma pneumoniae-specific antibody IgM, C-reactive protein, and procalcitonin levels in the blood of kids with Mycoplasma pneumoniae pneumonia as a reference for clinical diagnosis and treatment. The study group consisted of 96 children who had mycoplasma pneumonia in our hospital between May 2020 and May 2021, and the control group consisted of 96 healthy children who had a routine physical examination in our hospital at the same time. C-reactive protein and procalcitonin were measured and compared. The application value of single detection and combined detection of Mycoplasma pneumoniae-specific antibody IgM, C-reactive protein, and procalcitonin in the diagnosis of Mycoplasma pneumoniae pneumonia was evaluated based on clinical diagnosis results. The detection values of C-reactive protein and procalcitonin in the study group were higher than those in the recovery period and the control group, P < 0.05; the detection values of C-reactive protein and procalcitonin in the study group were higher than those in the control group, P < 0.05. The combination detection of Mycoplasma pneumoniae-specific antibody IgM, C-reactive protein, and procalcitonin had a greater diagnostic accuracy than single detection (P < 0.05). The sensitivity was higher than C-reactive protein and procalcitonin (P < 0.05); the specificity and positive predictive value were higher than Mycoplasma pneumoniae-specific antibody IgM (P < 0.05); and the negative predictive value was higher than procalcitonin (P < 0.05). The clinical value of combining the detection of Mycoplasma pneumoniae-specific antibody IgM, C-reactive protein, and procalcitonin in the diagnosis of Mycoplasma pneumoniae pneumonia in children is higher than that of single item detection, and it can provide a reliable clinical reference, as well as aid in evaluating the recovery effect of children, and it is worthy of application.

MAPK/MAK/MRK overlapping kinase mediated apoptosis through caspase signaling pathway from Cristaria plicata.

MAPK/MAK/MRK Overlapping Kinase (MOK) belongs to MAP kinase superfamily, which plays an important role in regulating cell growth, division, and differentiation. Caspase-3, as the final executor of apoptosis, has an important position in the caspase-mediated apoptotic signaling pathway. The full-length cDNA of MOK and caspase-3 were cloned from Cristaria plicata (designated CpMOK and CpCaspase-3). The CpMOK gene was sequence with a full-length of 1413 bp, encoding a total of 470 amino acids, and containing an S_TKc structural domain. CpCaspase-3 has a sequence of 2425 bp, encoding 322 amino acids, containing a CASc domain. Real-time fluorescence quantitative PCR analysis showed that CpMOK and CpCaspase-3 distributed in various tissues of C. plicata, and the highest expression of CpMOK and CpCaspase-3 mRNA was in hepatopancreas. The expression of CpMOK was significantly changed in hepatopancreas, gills, and kidneys by the construction of wound model as well as stimulation of LPS, PGN, Poly I: C and Aeromonas hydrophila. Subcellular localization experiments confirmed that CpMOK was localized in the nucleus. Furthermore, the double-stranded RNA (dsRNA) of CpMOK was constructed for interference experiment, and the results showed that the mRNA expression of apoptotic gene signals caspase-1, caspase-3, caspase-7, caspase-8, and caspase-9 were increased. The expression of caspase-1, -3, -7, -9, cytochrome C (Cyt-c) and tumor necrosis factor-α (TNF-α) was detected by ELISA. Fluorescent staining of apoptotic cells using the Tunnel method revealed an increase in the number of apoptotic cells after interference. These results suggested that CpMOK knockdown could induce caspase-mediated apoptosis in C. plicata, and the phosphorylation of the kinase was disrupted during the process.

Improving the quality of Suancai by inoculating with Lactobacillus plantarum and Pediococcus pentosaceus.

The quality characteristics of Suancai fermented with Lactobacillus plantarum CGMCC No.20193 (Lb. plantarum) and Pediococcus pentosaceus CGMCC No. 20192 (P. pentosaceus) were investigated. Their inoculation affected the bacterial communities revealed by Pacbio Sequel platform. After fermentation, the dominant phylum and genus in inoculation and spontaneous fermented Suancai were Firmicutes and Lactobacillus. Compared with single inoculation, the co-inoculation of Lb. plantarum and P. pentosaceus had a higher bacterial diversity. The Suancai co-inoculated with Lb. plantarum and P. pentosaceus had a more similar VCs profile with spontaneous fermented Suancai. The inoculation of Lb. plantarum and P. pentosaceus increased the content of organic acids, such as lactate, acetate, citrate, succinate, malate and tartrate. The most amino acids content in Suancai fermented with Lb. plantarum and P. pentosaceus were higher than that in spontaneous fermented Suancai. Compared single inoculation, the Suancai co-inoculated with Lb. plantarum and P. pentosaceus had a higher similarity of organoleptic tastes with spontaneous fermented Suancai. These results may facilitate the understanding of the starters' effects on the Suancai fermentation and the selection of applicable starters to manipulate the flavor.

Effects of salt concentration on the quality of paocai, a fermented vegetable product from China.

BACKGROUND: Paocai is a traditional Chinese fermented vegetable food. As the most important ingredient, salt has crucial effects on the bacterial community and volatile compounds of paocai. To demonstrate the effects of salt on the fermentation of paocai, the bacterial composition and volatile compounds were investigated using high-throughput sequencing and gas chromatography-mass spectrometry (GC-MS). RESULTS: The salt had no significant effects on the bacterial community at the phylum level. Proteobacteria and Bacteroidetes gradually decreased during the fermentation, and Firmicutes gradually increased as the dominant bacteria in the late stage of fermentation. At the genus level, Lactobacillus and Lactococcus gradually increased in relative abundance during the fermentation and became the dominant bacteria in paocai. High salt levels can contribute to the growth of Lactobacillus, which became the dominant genus in paocai. The salt concentration affected the profiles of volatile compounds in paocai after fermentation. A total of 42 volatile components were detected by GC-MS, among which phenols, aldehydes, and nitriles were the main ones. A high salt concentration will increase the volatile compound content, mainly aldehydes and alcohols, and improve the flavor of paocai. At the same time, the electronic tongue analysis also showed that a high salt concentration made a major contribution to the flavor of paocai. CONCLUSIONS: These data are helpful to elucidate the effects of salt on the quality of paocai and contribute to improving the quality and reducing the use of salt. © 2021 Society of Chemical Industry.

Changes in the expression of interleukin-10 in myocardial infarction and its relationship with macrophage activation and cell apoptosis.

BACKGROUND: Currently, the role of interleukin-10 (IL-10) as an anti-inflammatory factor in the occurrence and development of heart disease is still unclear. This study aimed to observe the dynamic changes in the expression of IL-10 in serum and myocardial tissues and to investigate the relationship of IL-10 expression with macrophage activation and cardiomyocyte apoptosis during the occurrence of myocardial infarction (MI). METHODS: Mice models with MI were prepared by ligating the anterior descending branch of the coronary artery. The animals were classified into the sham operation group (the control group), and the day 1, 7, 14, and 28 MI groups based. RESULTS: On days 7 and 14, the cells with positive IL-10 expression were largely distributed in the infarct areas, while cells with positive IL-10 expression were decreased on day 28. Serum IL-10 was significantly positively correlated with IL-10 protein expression in myocardial tissues. Moreover, Bcl-2 and Bax protein expression in myocardial tissues, along with the ratio of Bcl-2/Bax proteins, were gradually elevated with prolonged time of infarction. The expression of arginase protein increased gradually too. There were positive correlations between IL-10 and arginase expressions, and between the expressions of Bcl-2 and Bax proteins. CONCLUSIONS: After the occurrence of MI, the expression of IL-10 first increased and then decreased in serum and myocardial tissues, with this likely affecting macrophage activation, phenotypic transformation, and the occurrence of cardiomyocyte apoptosis.

Correlation between the trajectory of systolic blood pressure and new renal damage in a nonhypertensive population.

OBJECTIVE: This study aims to investigate the correlation between the trajectory of systolic blood pressure (SBP) and new renal damage in a nonhypertensive population. PATIENTS AND METHODS: This prospective cohort study included a total of 14 382 nonhypertensive individuals, employees of Kailuan Group of Companies, who took part in five healthy examinations in 2006-2007, 2008-2009, 2010-2011, 2012-2013, and 2014-2015, and had complete data. These individuals were divided into four groups according to the different trajectories of SBP: low-low, low-stable, middle-high, and high-high groups. The correlation between the trajectory of SBP and new renal damage in a nonhypertensive population was analyzed using a multivariate Cox's proportional hazard regression model. RESULTS: (a) A total of 14 382 individuals had complete data and the average age of these individuals was 44.6±10.8 years. Among these, 10 888 (75.7%) individuals were men and 3494 (24.3%) individuals were women. (b) These individuals were divided into four groups according to different trajectories of blood pressure: low-low group, accounting for 13.15% (blood pressure was <106 mmHg); low-stable group, accounting for 53.91% (blood pressure was between 115 and 116 mmHg); middle-high group, accounting for 28.77% (blood pressure was between 125 and 131 mmHg); and high-high group, accounting for 4.6% (blood pressure was between 126 and 151 mmHg). (c) With the increase in the trajectory of SBP, the detection rate of renal damage increased gradually. From the low-low group to the high-high group, the detection rates of estimated glomerular filtration rate (eGFR) less than 60 ml/min/1.73 m were 2.3, 2.4, 3.6, and 4.3%, respectively; the positive rates of urinary protein were 1.7, 2.9, 3.8, and 5.5%, respectively; and the detection rates of eGFR less than 60 ml/min/1.73 m or positive urinary protein were 4, 5.2, 7.3, and 9.3%, respectively (P<0.05). (d) After adjustment for other confounding factors, multivariate Cox's proportional hazard regression analysis showed that compared with the low-low group, the risk of eGFR less than 60 ml/min/1.73 m increased by nearly 1.5 times in the high-high group and in the low-stable, middle-high, and high-high groups, the risks of positive urinary protein, eGFR less than 60 ml/min/1.73 m, or positive urinary protein increased by 1.48-2.34 and 1.20-1.70 times, respectively. CONCLUSION: In a nonhypertensive population, the high trajectory of SBP is a risk factor for kidney damage.