Activation of dorsal horn cannabinoid CB2 receptor suppresses the expression of P2Y12 and P2Y13 receptors in neuropathic pain rats.

BACKGROUND: More evidence suggests that dorsal spinal cord microglia is an important site contributing to CB2 receptor-mediated analgesia. The upregulation of P2Y12 and P2Y13 purinoceptors in spinal dorsal horn microglia is involved in the development of pain behavior caused by peripheral nerve injury. However, it is not known whether the expression of P2Y12 and P2Y13 receptors at spinal dorsal horn will be influenced after CB2 receptor activation in neuropathic pain rats. METHODS: Chronic constriction injury (CCI) and intrathecal ADPbetaS injection were performed in rats to induce neuropathic pain. The paw withdrawal latency (PWL) was used to evaluate thermal hyperalgesia in neuropathic rats. The expression of P2Y12 and P2Y13 receptors, p-p38MAPK, and NF-kappaBp65 was detected with RT-PCR and western blotting analysis. RESULTS: Treatment with AM1241 produces a pronounced inhibition of CCI-induced thermal hyperalgesia and significantly inhibited the increased expression of P2Y12 and P2Y13 receptors at the mRNA and protein levels, which open up the possibility that P2Y12 and P2Y13 receptor expression are downregulated by CB2 receptor agonist AM1241 in CCI rats. Western blot analysis demonstrated that AM1241 reduced the elevated expression of p-p38MAPK and NF-κBp65 in the dorsal spinal cord induced by CCI. After administration with either SB203580 (p38MAPK inhibitor) or PDTC (NF-kappaB inhibitor), the levels of P2Y13 receptor expression in the dorsal spinal cord were lower than those in the CCI group. However, in CCI rats, the increased expression of P2Y12 receptor was prevented by intrathecal administration of PDTC but not by SB203580. In addition, minocycline significantly decreased the increased expression of P2Y12 and P2Y13 receptors. The similar results can be observed in ADPbetaS-treated rats. Intrathecal injection of ADPbataS causes thermal hyperalgesia and increased expression of P2Y12 and P2Y13 receptors in the dorsal spinal cord of naive rats. Moreover, intrathecal injection of AM1241 alleviates pain response and reduces the elevated expression of P2Y12 and P2Y13 receptors, p-p38MAPK, and NF-κBp65 in the dorsal spinal cord of ADPbetaS-treated rats. Intrathecal injection of SB203580 significantly inhibited the ADPbetaS-induced P2Y13 receptor expression, without affecting P2Y12 receptor expression. However, treatment with either SB203580 or PDTC effectively inhibited P2Y13 receptor expression compared to ADPbetaS-treated rats. CONCLUSIONS: In CCI- and ADPbetaS-treated rats, AM1241 pretreatment could efficiently activate CB2 receptor, while inhibiting p38MAPK and NF-kappaB activation in the dorsal spinal cord. CB2 receptor stimulation decreased P2Y13 receptor expression via p38MAPK/NF-kappaB signaling. On the other hand, CB2 receptor activation decreased P2Y12 receptor expression via p38MAPK-independent NF-kappaB signaling pathway.

Seismic Behavior Analysis of Recycled Aggregate Concrete-Filled Square Steel Tube Frames.

In this study, the seismic behavior of a recycled aggregate concrete-filled square steel tube (S-RACFST) frame under different design conditions was investigated. Based on previous studies, a finite element model for the seismic behavior of the S-RACFST frame was developed. Moreover, the axial compression ratio, beam-column line stiffness ratio, and yield bending moment ratio of the beam-column were regarded as the variation parameters. It was through these parameters that the seismic behavior of eight S-RACFST frame finite element specimens was discussed. The seismic behavior indexes, such as the hysteretic curve, ductility coefficient, energy dissipation coefficient, and stiffness degradation were obtained-which, in turn, revealed the influence law and the degree of the design parameters regarding seismic behavior. Moreover, the sensitivity of the various parameters with respect to the seismic behavior of the S-RACFST frame was evaluated via grey correlation analysis. The results show that the hysteretic curves of the specimens were fusiform and full with respect to the different parameters. Firstly, with the axial compression ratio increasing from 0.2 to 0.4, the ductility coefficient increased by 28.5%. In addition, the equivalent viscous damping coefficient of the specimen with the axial compression ratio of 0.4 was 17.9% higher than that of the specimen with the axial compression ratio of 0.2, which was 11.5% as well as that with an axial compression ratio of 0.3. Second, when the line stiffness ratio rises from 0.31 to 0.41, the specimens' bearing capacity and displacement ductility coefficient both get better. However, the displacement ductility coefficient gradually decreases when the line stiffness ratio is greater than 0.41. As a result, an optimal line stiffness ratio (0.41) thus exhibits good energy dissipation capacity. Thirdly, with the increase in the yield bending moment ratio from 0.10 to 0.31, the bearing capacity of the specimens improves. In addition, the positive and negative peak loads increased by 16.4% and 22.8%, respectively. Moreover, the ductility coefficients were all close to three, thus demonstrating good seismic behavior. The stiffness curve of the specimen with a large yield bending moment ratio with respect to the beam-column, is higher than those that possess a small beam-column yield moment ratio. In addition, the yield bending moment ratio of the beam-column possesses a significant influence on the seismic behavior of the S-RACFST frame. Furthermore, the yield bending moment ratio of the beam-column should be considered first in order to ensure the seismic behavior of the S-RACFST frame.

Isolation of Cytochrome C for Proteomics with Lindqvist-type Polyiodate Modified Metal Organic Framework.

Separation and purification of Cytochrome C (Cyt-C) is important for proteomic. High efficient and selective pretreatment method for Cyt-C in real samples are always needed. Herein, polyniobate (K7H[Nb6O19]·13H2O, Nb6O19) is modified on a metal-organic framework MIL-125(Ti) through intermolecular hydrogen bonds and an aqueous-stable composite Nb6O19/MIL-125(Ti) is successfully prepared to pretreat complex protein sample. Protein adsorption studies have shown that Nb6O19/MIL-125(Ti) can promote the selective adsorption of Cyt-C due to the synergistic effect of electrostatic and hydrogen-bond interactions. At pH=10.0 (Britton-Robinson buffer), the adsorption efficiency of 300 µL 100 µg·mL-1 Cyt-C onto 1.0 mg Nb6O19/MIL-125(Ti) can reach 99.5%. The adsorption behavior of Cyt-C fits well with the Langmuir adsorption model, corresponding to a maximum theoretical adsorption capacity of 168.35 mg·g-1. Using 3 mol·L-1 NaCl as the eluent, a high elution efficiency of 92.19% is obtained. In addition, the results of the sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis confirm that Nb6O19/MIL-125(Ti) efficiently adsorbed Cyt-C from scrofa heart extraction. LC-MS/MS spectrometry results show that the purification of Cyt-C reduces the abundance from the 12th to the 154th place after Nb6O19/MIL-125(Ti) treatment. Moreover, low abundant proteins, e.g., Superoxide dismutase 1, IF rod domain-containing protein and Ubiquitin-60S ribosomal protein L40 were considerably enriched. These outcomes confirm the practicability of Nb6O19/MIL-125 (Ti) as a Cyt-C extractant has potential application value in scrofa heart proteomics.

Outcome of Nursing Based on Health Belief United with Knowledge, Belief, and Practice Mode on Gastroscopy of Patients with Gastric Cancer.

Aim: If gastric cancer can be detected through early screening, and scientific and reasonable intervention methods can be selected in time, the condition can be effectively controlled. Routine nursing has been unable to obtain satisfactory results, and the effect on improving the compliance of the examiner is not outstanding. The research aims to estimate the outcome of nursing based on health belief combined with knowledge, belief, and practice on gastroscopy in patients with gastric cancer. Methods: 126 patients with clinically diagnosed gastric cancer in the Number Two Hospital of Baoding from May 2020 to May 2022 were randomly divided into belief guidance group and mode group, with 63 instances each. The mode group was intervened via the mode of knowledge, belief, and practice, and the belief guidance group was intervened via the nursing based on health belief on the basis of the mode group. Before and after the nursing, the health belief, examination compliance, inappropriateness, and negative emotion in different time periods were contrasted between the two groups. Results: After the nursing, the scores of health belief scale in the belief guidance group were enhanced than those in the mode group; the compliance rate of the belief guidance group was markedly enhanced than that of the mode group, and the inappropriateness during the insertion and examination was lower than that of the mode group; the scores of self-rating anxiety scale (SAS) and self-rating depression scale (SDS) in the two groups preinsertion and postnursing were markedly lower than those in the mode group. Conclusion: Nursing based on health belief guidance united with knowledge, belief, and practice mode nursing can advance the health belief and compliance of gastroscopy in patients with gastric cancer, reduce discomfort, and effectively advance the negative emotions of patients. It is worthy of clinical application.

Application of artificial neural networks to predict multiple quality of dry-cured ham based on protein degradation.

This study investigated protein degradation and quality changes during the processing of dry-cured ham, and then established the multiple quality prediction model based on protein degradation. From the raw material to the curing period, proteolysis index of external samples were higher than that of internal samples, however, the difference gradually decreased from the drying period to the maturing period. Protein degradation can be used as indicators for controlling quality of the hams. With protein degradation index as input variables, the back propagation-artificial neural networks (BP-ANN) models were optimized, with training function of trainlm, transfer function of logsig in input-hidden layer and tansig in hidden-output layer, and 20 hidden layer neurons. Furthermore, the relative errors of predictive data and experimental data of 12 samples were approximately 0 with the BP-ANN model. Results indicated that the BP-ANN has great potential in predicting multiple quality of dry-cured ham based on protein degradation.

Substituting sodium by various metal ions affects the cathepsins activity and proteolysis in dry-cured pork butts.

Salt contents and compositions are crucial parameters to enzyme activity and might even affect the proteolysis and quality of dry-cured meat. However, the metal ions significantly vary with geographic origin, which would be a determinant in the dry-cured meat manufacture. Therefore, the main salt compositions of KCl, CaCl2, MgCl2 and ZnCl2 were therefore used to partially substitute NaCl while additionally assessing and regulating their function during the dry-cured pork butts processing. Physiochemical properties, cathepsins activity, proteolysis and sensory evaluation were investigated. The results indicated that partial substitution of sodium prevented the dehydration, and accelerated the pH reduction. CaCl2 and MgCl2 partial substitution moreover exerted some promoting effect on cathepsin activity whereas ZnCl2 was a subtle inhibitor. The proteolysis index of CaCl2 and MgCl2 substitution were superior to the rest. The metal ions partial substitution reduced saltness, while the presence of KCl and MgCl2 enhanced bitterness. Further correlation analysis was performed to better understand the interactions between those parameters.

A rapid and reliable multiplex PCR assay for simultaneous detection of fourteen animal species in two tubes.

A simple and rapid method for animal species identification to prevent food adulteration based on mitochondrial DNA using two independent multiplex polymerase chain reactions (PCRs) and microchip electrophoresis was developed. This method was designed to identify fourteen domestic animals (Group I: cattle, donkey, dog, fox, raccoon-dog, deer and horse; Group II: pig, sheep, goat, chicken, duck, cat and mouse) simultaneously using ten pairs of primers and three of which were degenerate primers. Sequences for species-specific primers were generated based on mitochondrial genes, including 12S rRNA, 16S rRNA, ND2 and CO I. This method was validated in terms of the specificity, sensitivity and practicability, and the developed multiplex PCR method was able to correctly identify animal species of raw meats and processed meat products. The detection limits of two multiplex PCRs were 0.02 ng DNA for animal species in Group I and 0.2 ng DNA for Group II, respectively.

P2Y6 receptor activation is involved in the development of neuropathic pain induced by chronic constriction injury of the sciatic nerve in rats.

Purinergic signaling in spinal cord microglia plays an important role in the pathogenesis of neuropathic pain. Among all P2 receptors, P2Y6 receptor is expressed in rat dorsal spinal cord. However, it's not clear that the role of P2Y6 receptor in the chronic constriction injury (CCI) model of neuropathic pain rats. We evaluated the effect of repeated intrathecal administration of MRS2578 (selective P2Y6 receptor antagonist) on CCI-induced nociceptive behaviors in rats. After CCI, MRS2578 (10-11-10-4 M) was administration. The thermal withdrawal latency (TWL) and mechanical withdrawal threshold (MWT) were assessed. The expression of P2Y6 receptor and Iba-1 at rat dorsal spinal cord was observed by using RT-PCR. We found that intrathecal injection of MRS2578 suppressed CCI-induced mechanical allodynia and thermal hyperalgesia with a dose-dependent manner. The CCI rats presented increased expression of P2Y6 receptor and Iba-1 at the mRNA level in the ipsilateral dorsal spinal cord than that in sham group. Treatment with either minocycline or SB203580 effectively inhibited P2Y6 receptor expression compared to CCI rats. Intrathecal injection of UDP enhanced mechanical and thermal allodynia than that in CCI group. To the further study, intrathecal injection of UDP causes mechanical allodynia and thermal hyperalgesia in naive rats. The increased expression of P2Y6 receptor and Iba-1 were observed in UDP-treated rats. Intrathecal injection of MRS2578 alleviates pain response in UDP-treated rats. These observations suggested that P2Y6 receptor in dorsal spinal cord contribute to mechanical allodynia and thermal hyperalgesia in CCI-induced neuropathic pain.

N-fixing trees in wetland restoration plantings: effects on nitrogensupply and soil microbial communities.

To investigate the impact of an exotic Frankia nodulated tree (Alnus trabeculosa) on soil nitrogen content, soil microbial composition, and the abundance of N turnover-related functional microorganism community, we compared the community structure and abundance of key functional genes (nifH, bacterial/archaeal amoA, and nosZ) in the rhizosphere and nonrhizosphere of monoculture of Phragmites australis and A.trabeculosa-P.australis mixed communities by MiSeq Illumina sequencing and real-time PCR, respectively. The introduction of Frankia nodulated tree to recover degraded wetland was effective in the accumulation of soil organic carbon and nitrogen, which was the key factor to impact on the bacterial community composition revealed by canonical correspondence analysis. Acidobacteria and Proteobacteria were the dominant bacterial phylums while seven rare phyla appeared the most phylogenetically different among the investigated soil of two vegetations, including Chlorobi, Cyanobacteria, OD1, OP11, TM6, TM7, and GN02. The gene copy numbers of nifH were ranged from 2.28 × 108 to 2.96 × 109 copies g-1 dry soil in the wetland, and which were significantly higher in soil samples from P. australis than that from A.trabeculosa. While the abundance of nosZ in both rhizosphere and nonrhizosphere soils of A.trabeculosa-P.australis mixed communities was significantly lower compared with P.australis monoculture. The potential nitrification (PNA) (0.15-0.41 mg NOx-N kg-1 dry soil d-1) in the rhizosphere of A. trabeculosa was significantly higher than that of P. australis, and the soil denitrification enzyme activity (DEA) (0.42-0.90 nmol N2O-N g-1 dry soil h-1) was lower in the mixed community compared with monoculture of P. australis. The introduced planting of Frankia nodulated tree effectively accumulated soil organic carbon and nitrogen and reduce the relative abundance and activity of nitrogen-fixing bacteria and denitrification bacteria.

Two efficient nitrite-reducing Lactobacillus strains isolated from traditional fermented pork (Nanx Wudl) as competitive starter cultures for Chinese fermented dry sausage.

Lactic acid bacteria isolated from traditional Dong pork product (Nanx Wudl) were investigated for their potential as starter cultures for Chinese fermented dry sausages. Based on preliminary screening, Lactobacillus plantarum CMRC6 and Lactobacillus sakei CMRC15, both showing excellent nitrite-reducing capacity, were used as single-strain starter cultures. For comparison, a commercial composite starter was also tested. In CMRC6 and CMRC15-inoculated sausages, lactic acid bacteria dominated the microflora and improved the microbiological safety by suppression of Enterobacteriaceae growth. Nitrite content of all inoculated sausages declined rapidly during ripening compared to non-inoculated. Texture profiles analysis showed inoculated sausages had more pronounced textural development during ripening. Sensory evaluation indicated CMRC6 and CMRC15-fermented sausages had comparable or more desirable organoleptic characteristics than sausage made with commercial starters. Therefore, CMRC6 and CMRC15 are promising candidates as multi-functional starter cultures for microbiological safety and residual nitrite control in gourmet Chinese dry sausage production.