Deciphering the key factors affecting pesticide residue risk in vegetable ecosystem.

Soil contamination, particularly from pesticide residues, presents a significant challenge to the sustainable development of agricultural ecosystems. Identifying the key factors influencing soil pesticide residue risk and implementing effective measures to mitigate their risks at the source are essential. Here, we collected soil samples and conducted a comprehensive survey among local farmers in the Three Gorges Reserve Area, a major agricultural production region in Southwest China. Subsequently, employing a dual analytical approach combining structural equation modeling (SEM) and random forest modeling (RFM), we examined the effects of various factors on pesticide residue accumulation in vegetable ecosystems. Our SEM analysis revealed that soil characteristics (path coefficient 0.85) and cultivation factor (path coefficient 0.84) had the most significant effect on pesticide residue risk, while the farmer factors indirectly influenced pesticide residues by impacting both cultivation factors and soil characteristics. Further exploration using RFM identified the three most influential factors contributing to pesticide residue risk as cation exchange capacity (CEC) (account for 18.84%), cultivation area (account for 14.12%), and clay content (account for 13.01%). Based on these findings, we carried out experimental trials utilizing Integrated Pest Management (IPM) technology, resulting in a significant reduction in soil pesticide residues and notable improvements in crop yields. Therefore, it is recommended that governmental efforts should prioritize enhanced training for vegetable farmers, promotion of eco-friendly plant protection methods, and regulation of agricultural environments to ensure sustainable development.

BMP-2 promotes osteogenic differentiation of mesenchymal stem cells by enhancing mitochondrial activity.

OBJECTIVES: Mesenchymal stem cells (MSCs) have become seed cells and basic elements for bone regeneration and bone tissue engineering. The aim of the present study was to investigate the roles and mechanisms of bone morphogenetic protein 2 (BMP-2) on osteogenic differentiation of MSCs. METHODS: Primary MSCs were isolated from the femur and tibia bone of rats and then transfected with BMP-2 and PGC-1α adenovirus vectors. Alkaline phosphatase (ALP) activity and alizarin red staining were used to measure osteogenic differentiation of MSCs. Real-time PCR and western blot assays were performed to assess osteogenic differentiation-related proteins levels. The activities of mitochondrial respiratory chain complexes I and II and mitochondrial fluorescence intensity were used to explore mitochondria status during osteogenic differentiation of MSCs. RESULTS: We found that the ability of BMP-2 overexpressed (OE) group osteogenic differentiation was significantly improved, compared with the negative control (NC) group. The results also indicated that BMP-2 can promote the activity of mitochondria. We further used the gain- and loss-of-function approaches to demonstrate that BMP-2 promotes mitochondrial activity by up-regulating PGC-1α to promote osteogenic differentiation of MSCs. CONCLUSIONS: These results explored the important role of BMP-2 in the osteoblast differentiation of MSCs from a new perspective, providing a theoretical and experimental basis for bone defect and repair.

Role of tumor necrosis factor receptor­associated factor 6 in pyroptosis during acute pancreatitis.

Acute pancreatitis (AP) is hypothesized to be related to the activation of an inflammatory response induced by pyroptosis. The aim of the present study was to investigate the potential role of tumor necrosis factor receptor­associated factor 6 (TRAF6) in pyroptosis in an AP rat model and the human pancreatic ductal epithelial HPDE6C7 cell line. In vivo, AP was induced by intraperitoneal injection of caerulein (CAE) in rats. The rats were sacrificed at 24 or 48 h after the final CAE injection. In vitro, HPDE6C7 cells were treated with CAE for 12, 24 and 48 h. Moreover, TRAF6 was overexpressed and treated with CAE for 48 h. Histopathological changes of pancreatic, serum and supernatant inflammatory cytokines and pyroptosis­related mRNA and protein expression levels were determined by histopathological scores, ELISA, reverse transcription­quantitative PCR and western blotting. In addition, pyroptosis morphological changes were also determined by Hoechst/PI staining in HPDE6C7 cells. Results showed that AP was observed in the CAE­induced rat model, and that serum IL­1ß and IL­18 levels, and TRAF6, NLR pyrin domain containing 3 (NLRP3), caspase­1 and caspase­3 mRNA and protein expression levels were increased. Similar in HPDE6C7 cells, CAE treatment caused supernatant IL­1ß level, NLRP3 and caspase­1 mRNA expression levels to significantly increase. After TRAF6 overexpression and CAE treatment, supernatant IL­1ß level, caspase­1 protein expression level, and NLRP3 and caspase­3 mRNA and protein expression levels were also significantly increased. Furthermore, cells exhibited red fluorescence in Hoechst/PI staining, which can be used as a method of detecting pyroptosis activation. The results also showed that the red fluorescence was stronger after CAE treatment or TRAF6 overexpression plus CAE treatment. In conclusion, TRAF6 and caspase­1/3 signaling pathways were involved in the pathogenesis of CAE­induced AP in rats. Pyroptosis was activated by CAE and TRAF6 overexpression via the caspase­1/3 signaling pathways in HPDE6C7 cells.

Activation of the Nuclear Factor-kappa B Signaling Pathway Damages the Epithelial Barrier in the Human Pancreatic Ductal Adenocarcinoma Cell Line HPAF-II.

OBJECTIVES: Injury of the pancreatic duct epithelial barrier plays a critical role in the development of acute pancreatitis. The activity of the nuclear factor-kappa B (NF-κB) pathway is involved in the disruption of the pancreatic duct epithelial barrier. This study investigated how NF-κB impacts the dysfunction of the pancreatic duct epithelial barrier. METHODS: A human pancreatic ductal adenocarcinoma cell line was treated with tumor necrosis factor-alpha (TNF-α) and pyrrolidine dithiocarbamate. The expression levels of p65 and p-p65 were detected to evaluate NF-κB activity. Tricellulin (TRIC) expression levels were measured to assess the change in tight junction (TJ)-related proteins. The expression and localization of myosin light chain kinase (MLCK) were investigated. The structure of TJs and monolayer permeability were also examined. RESULTS: NF-κB was activated by TNF-α and suppressed by pyrrolidine dithiocarbamate. Activation of NF-κB upregulated the expression levels of TRIC and MLCK. Broadened TJs were observed after NF-κB was activated. Lower monolayer permeability was observed when NF-κB was suppressed. CONCLUSIONS: Activation of the NF-κB pathway induced by TNF-α leads to increased TRIC and MLCK expression, resulting in broadened TJs and high permeability, which contribute to damage to the pancreatic duct epithelial barrier.

Comparison of the effects of minimally invasive percutaneous pedicle screws osteosynthesis and open surgery on repairing the pain, inflammation and recovery of thoracolumbar vertebra fracture.

We compared the clinical effects of minimally invasive percutaneous pedicle screws osteosynthesis (MIPPSO) and open surgery on the repair of thoracolumbar vertebra fracture. Seventy patients, who suffered from thoracolumbar vertebra fracture and received treatment at our hospital, were selected and randomly divided into either the minimally invasive percutaneous pedicle screws osteosynthesis group (MIPPSO group) and the traditional open pedicle screws osteosynthesis group (TOPSO group) with 35 cases in each group. The perioperative parameters including length of incision, duration of operation, bleeding during operation, length of hospital stay, the changes of pre-operative and post-operative VAS pain scores, inflammatory indexes including serum C-reactive protein (CRP) and creatine kinase (CK), and imaging indexes including Cobb's angle and anterior margin height of vertebral body of both groups were compared. The length of incision, duration of operation, bleeding volume during operation and length of hospital stay in the MIPPSO group were significantly lower than those in the TOPSO group (P<0.05). Both minimally invasive surgery and traditional surgery effectively alleviated the pain (P<0.05), which was more significant in MIPPSO group (P<0.05). The post-operative inflammatory indexes, CRP and CK levels, of both groups were higher compared to the pre-operation (P<0.05), which was more significant in TOPSO group (P<0.05). The differences of imaging indexes, including Cobb's angle and anterior margin height of injured vertebra, were statistically significant between pre-operation and post-operation for each group (P<0.05); however, there were no statistically significant differences between two groups at either pre-operation or post-operation (P>0.05). The effect of minimally invasive percutaneous pedicle screws osteosynthesis is similar to the traditional open surgery, however, the MIPPSO technique has the advantages of small trauma, less bleeding, short duration of operation, rapid post-operative recovery, light pain, less economic cost, and better aesthetic effect and is therefore worthy of clinical promotion.

Isolation and engineering of plant growth promoting rhizobacteria Pseudomonas aeruginosa for enhanced cadmium bioremediation.

Although many bacteria are tolerant to heavy metals and play important roles in the immobilization of heavy metals, they cannot always be dependably reproduced under field conditions. In this work, a cadmium (Cd)-resistant bacterium was isolated from a Cd-contaminated oil field and identified as Pseudomonas aeruginosa (Pse-w). We then determined various plant growth promoting features such as the solubilization of phosphate, and the production of indole-3-acetic acid and siderophores. Lastly, we engineered the strain Pse-w-MT by targeting metallothioneins to the cell surface of Pse-w to immobilize Cd2+ and promote plant growth. Our data revealed that Pse-w exhibited high levels of resistance to Cd2+ (4 mM) and showed various plant growth promoting features. The engineered strain Pse-w-MT was found to adsorb Cd2+ mainly via extracellular deposition, and had an enhanced ability for immobilizing Cd2+ ions from the external media. Furthermore, the inoculation of Cd-polluted soil with Pse-w-MT significantly elevated the shoot and root biomass and leaf chlorophyll content. Similarly, plants inoculated with Pse-w-MT demonstrated markedly lower Cd2+ accumulation in the root and shoot system. It was concluded that plant growth promoting rhizobacteria with a high Cd2+ tolerance was an ideal candidate to be engineered for bioremediation and plant growth promotion against Cd-induced stress.

Achieving High Yield of Lactic Acid for Antimicrobial Characterization in Cephalosporin-Resistant Lactobacillus by the Co-Expression of the Phosphofructokinase and Glucokinase.

Lactobacilli are universally recognized as probiotics that are widely used in the adjuvant treatment of inflammatory diseases, such as vaginitis and enteritis. With the overuse of antibiotics in recent years, the lactobacilli in the human body are killed, which could disrupt the microecological balance in the human body and affect health adversely. In this work, cephalosporin-resistant Lactobacillus casei RL20 was obtained successfully from the feces of healthy volunteers, which possessed a stable genetic set. However, the shortage of lactic acid (72.0 g/l at 48 h) by fermentation did not meet the requirement for its use in medicine. To increase the production of lactic acid, the functional genes pfk and glk were introduced into the wild strain. A yield of 144.2 g/l lactic acid was obtained in the transgenic L. casei RL20-2 after fermentation for 48 h in 1 L of basic fermentation medium with an initial glucose concentration of 100 g/l and increasing antibacterial activity. These data suggested that L. casei RL20-2 that exhibited a high yield of lactic acid may be a potential probiotic to inhibit the spread of bacterial infectious diseases and may be used for vaginitis therapy.

Oral delivery of Bifidobacterium longum expressing α-melanocyte-stimulating hormone to combat ulcerative colitis.

α-Melanocyte-stimulating hormone (α-MSH) is a tridecapeptide derived from pro-opiomelanocortin that exhibits potent anti-inflammatory properties by regulating the production of inflammatory mediators. This peptide has been well established in several inflammatory models, including inflammatory bowel disease (IBD). However, its extremely short duration in vivo limits its clinical application. To address this limitation, Bifidobacterium was used here as a carrier to deliver α-MSH. We utilized α-MSH-engineered Bifidobacterium against IBD, which is closely linked to immune and intestinal microbiota dysfunction. First, we constructed a Bifidobacterium longum secreting α-MSH (B. longum-α-MSH). We then tested the recombinant α-MSH expression and determined its bioactivity in HT-29 cells. To assess its effectiveness, B. longum-α-MSH was used against an ulcerative colitis (UC) model in rats induced by dextran sulfate sodium. The data showed that α-MSH expression in B. longum-α-MSH was effective, and its biological activity was similar to the synthesized one. This UC model experiment indicated that B. longum-α-MSH successfully colonized the intestinal gut, expressed bioactive α-MSH and had a significant anti-inflammatory effect. The results demonstrate the feasibility of preventing IBD by using B. longum-α-MSH.

Oral Bifidobacterium longum expressing alpha-melanocyte-stimulating hormone to fight experimental colitis.

The oral delivery of peptides is a highly attractive treatment approach. However, the harsh environment of the gastrointestinal tract limits its application. Here, we utilize Bifidobacterium as a delivery system to orally deliver a potent anti-inflammatory but short duration peptide alpha-melanocyte-stimulating hormone (α-MSH) against experimental colitis. The aim of our study was to facilitate the efficient oral delivery of α-MSH. We designed a vector of pBDMSH and used it to construct a Bifidobacterium longum expressing α-MSH. We then determined the bioactivity of recombinant Bifidobacterium in lipopolysaccharide-induced inflammatory models of HT-29 cells. Finally, we used Bifidobacterium expressing α-MSH against dextran sulfate sodium (DSS)-induced ulcerative colitis mice. Results based on the myeloperoxidase activity, the levels of inflammatory cytokines TNF-α, IL-1ß, IL-6, and IL-10 and the histological injury of colon tissue reveal recombinant Bifidobacterium was efficient in attenuating DSS-induced ulcerative colitis, suggesting an alternative way to use Bifidobacterium as a delivery system to deliver α-MSH for DSS-induced ulcerative colitis therapy.

HIV-1 Coreceptor CXCR4 Antagonists Promote Clonal Expansion of Viral Epitope-Specific CD8+ T Cells During Acute SIV Infection in Rhesus Monkeys In Vivo.

BACKGROUND: The underlying molecular mechanisms and the kinetics of T cell receptor (TCR) repertoire selection during administration of CXCR4 or CCR5 inhibitors in infection of AIDS viruses in vivo have remained largely unexplored. Viral epitope-specific CD8(+) T lymphocytes play a dominant role in the control of HIV and simian immunodeficiency virus (SIV). We hypothesized that blockade of CXCR4 or CCR5 might influence the clonal expansion of epitope-specific CD8(+) T cells, contributing to antiviral immune responses in vivo. METHODS: We measured frequencies of the dominant epitope p11C-specific CD8(+) T cells and analyzed the TCR repertoire of those cells in SIV-infected rhesus monkeys treated by CXCR4 or CCR5 inhibitors and vMIP-II, which binds multiple chemokine receptors. RESULTS: A significantly increase in the levels of epitope-specific CD8(+) T cells was observed after blockade of CXCR4 or CCR5 compared with untreated control groups. Those CD8(+) T cells exhibited selected usage of TCR Vß families and complementarity-determining region 3 (CDR3) segments. The clonal expansion of distinct Vß populations could efficiently inhibit SIV replication in vitro, and CXCR4 inhibitor induced more expansion of epitope-specific CD8(+) T cells than CCR5 antagonist (P < 0.01), whereas vMIP-II treatment showed the most marked augmentation of p11C-specific CD8(+) T cells. CONCLUSIONS: Antagonists of HIV coreceptors, particularly CXCR4, play an important role in the clonal expansion of SIV epitope-specific CD8(+) T cells in vivo, thus inhibitors of chemokine receptors such as CXCR4 or CCR5 may contribute to the ability of epitope-specific CD8(+) T cells to inhibit SIV or HIV infection.